Clinicopathological features of very well-differentiated adenocarcinoma of the stomach / 中华病理学杂志

Objective: To investigate the clinicopathological features of very well-differentiated adenocarcinoma (VWDA) of the stomach. Methods: The clinicopathological data of 12 cases of VWDA of the stomach were collected retrospectively at the People's Liberation Army Joint Logistics Support Force 989 Hospital (formerly 152 Hospital), Pingdingshan, China, from January 2013 to May 2021. The histological characteristics and immunophenotypes were observed and analyzed with review of current literature. Results: There were 8 males and 4 females with a median age of 63 years (range 47 to 80 years). The tumor involved in the upper part of the stomach in 6 cases, the middle part in 2 cases, and the lower part in 4 cases. The median diameter of the tumors was 17 mm (range 5-65 mm). The tumor cells were similar to absorbent cells, Paneth cells, foveolar epithelial cells, and goblet cells. The cells were arranged in a single layer, and the nuclei were slightly enlarged and located at the base. The nuclei were fusiform to slightly irregular, with loss of nuclear polarity. Early tubular VWDA was found in 9 cases, and the tumor glands were similar to intestinal metaplasia. In two cases the tumors infiltrated into the submucosa. The lesions in the mucosa and submucosa showed the glands with cystic expansion, bending, branching, spiky and abortive growth pattern. One case of early papillary tubular VWDA was confined to the mucosal layer and composed of foveolar-type epithelial cells. There were two cases of advanced papillary tubular VWDA, which consisted of foveolar-type epithelial, pyloric glands, or mucinous neck cells and were associated with intra-lymphatic cancer embolus and lymph node metastases. Background mucosal atrophy and intestinal metaplasia were observed in all cases. Immunohistochemical staining showed intestinal type VWDA in 1 case, mixed gastrointestinal type VWDA in 9 cases, and gastric type VWDA in 2 cases. The Ki-67 proliferation index of 8 cases limited to the mucosa was 40%-70%, 2 cases of infiltration into the submucosa and 2 cases of advanced carcinoma was 10%-25%. All the tumors showed a wild type of p53 protein expression pattern and negative HER2. Adenocarcinoma or high-grade dysplasia was diagnosed on preoperative biopsy in 5 cases, and chronic atrophic gastritis with intestinal metaplasia in 7 cases. The median follow-up time was 28 months (range 12-72 months). No recurrence was found in the 10 patients with early cancer. Of the two patients with advanced carcinoma, one patient had lung metastases and the other died. Conclusions: Gastric VWDA is a rare low-grade malignancy with structural features of highly differentiated adenocarcinoma and extremely low cytological atypia. The diagnostic value of structural abnormality is significantly greater than cytological atypia. The invasive growth of irregular glands in the deep mucosa and submucosa is reliable evidence for diagnosis. The diagnosis of intramucosal VWDA is challenging and very difficult in some biopsy specimens.

Expression of silent information regulator 1 and 3 and hypoxia-inducible factor 1α in cutaneous squamous cell carcinoma tissues and cells / 中华皮肤科杂志

Objective:To determine the expression of silent information regulator 1 (Sirt1) , Sirt3 and hypoxia-inducible factor 1α (HIF-1α) in cutaneous squamous cell carcinoma (CSCC) tissues and cells, and to explore their role in the occurrence and development of CSCC.Methods:From January 2019 to December 2020, 30 lesional skin tissues were obtained from patients with histopathologically confirmed poorly-, moderately- or well-differentiated CSCC, and 30 normal skin tissues were obtained from patients with non-cancerous diseases in Department of Dermatology, General Hospital of Ningxia Medical University. A CSCC cell line A431 and a human keratinocyte cell line HaCaT were cultured. Immunohistochemical study, Western blot analysis and real-time quantitative PCR (RT-PCR) were performed to determine the protein and mRNA expression of Sirt1, Sirt3 and HIF-1α in CSCC tissues of different grades of differentiation and normal skin tissues, cytochemical and immunofluorescence staining and RT-PCR were conducted to determine the protein and mRNA expression of Sirt1, Sirt3 and HIF-1α in A431 and HaCaT cells, respectively. Comparisons of measurement data among multiple groups were performed by using one-way analysis of variance, and comparisons between two groups by using t test. Results:Immunohistochemical study showed that the expression level of Sirt3 (expressed as the average optical density) was 100 ± 12.12, 117.72 ± 26.23, 127.32 ± 24.45, 132.71 ± 31.61 in the normal skin tissues and well-, moderately- and poorly-differentiated CSCC tissues respectively, and there was a significant difference among these groups ( F = 20.14, P < 0.001) ; the expression of Sirt1 and HIF-1α increased in turn from the normal skin tissues to the well-, moderately- and poorly-differentiated CSCC tissues, and significantly differred in these groups ( F = 174.50, 225.00, respectively, both P < 0.001) . As Western blot analysis revealed, the expression level of Sirt3 significantly differed among the normal skin tissues, well-, moderately- and poorly-differentiated CSCC tissues (expressed as relative gray value: 1.000 ± 0.132, 1.403 ± 0.411, 1.387 ± 0.393, 1.677 ± 0.683, respectively; F = 34.97, P < 0.001) , and so did the expression levels of Sirt1 and HIF-1α ( F = 69.29, 199.90, respectively, both P < 0.00l) , with a gradually increasing trend in their expression levels from the the normal skin tissues to well-, moderately- and poorly-differentiated CSCC tissues. RT-PCR showed that the mRNA expression of Sirt3, Sirt1 and HIF-1α was sequentially increased from the normal skin tissues to well-, moderately- and poorly-differentiated CSCC tissues, and significant differences were observed among these groups ( F = 113.00, 174.50, 50.33, respectively, all P < 0.001) . The protein expression levels of Sirt3, Sirt1 and HIF-1α were significantly higher in the A431 cells than in the HaCaT cells ( t = 16.75, 18.34, 27.76, respectively, all P < 0.001) , and so were their mRNA expression levels ( t= 14.22, 9.62, 16.86, respectively, all P < 0.001) . Conclusion:Increased expression of Sirt3, Sirt1 and HIF-1α was observed in CSCC tissues and cells, which may promote the occurrence and development of CSCC.

Molecular identification and quality analysis of Rhei Rhizoma based on chloroplast matK gene and UPLC / 药学学报

Rhei Rhizoma is commonly used as a traditional Chinese medicine with multiple botanical origins. Different botanical sources may have different pharmacological activities. The germplasm resources of commercial Rhei Rhizoma were determined based on the chloroplast gene matK, and the anthraquinone and free anthraquinone content was determined by UPLC to analyze quality of commercial Rhei Rhizoma. Eighty-nine commercial Rhei Rhizoma samples were collected from 40 cities in 27 provinces. DNA was extracted and the matK gene was amplified by PCR. Results indicated that the collected samples were from the same botanical origin, Rheum palmatum, and 8 genotypes were identified, including Rp1, Rp2, Rp3, Rp4, Rp5, Rp6, Rp10 and Rp12. Rp4 and Rp6, cultivated in Gansu, Sichuan and Yunnan provinces were the main circulating genotypes, representing 40.45% and 37.08% of the total samples, respectively. Phylogenetic tree analysis showed that the eight genotypes were mainly divided into two branches, of which the main genotypes Rp4 and Rp6 were in one branch. Genetic distance analysis indicated that the genetic separation of the eight genotypes was between 0.001 and 0.010. UPLC analysis indicated that 93.26% of the samples met the Pharmacopoeia standards. There were significant differences in the content of total anthraquinone and free anthraquinone among the samples, in which the difference in free anthraquinone was 1.01% and the difference in total anthraquinone content was 3.79%, indicating that the quality of commercial Rhei Rhizoma samples varies considerably. There was no significant difference in the content of total anthraquinone and free anthraquinone in commercial Rhei Rhizoma among different collection provinces and genotypes. This study will help guide the circulation of Rhei Rhizoma in the market and provides valuable insights for molecular identification and quality analysis of other traditional Chinese medicines.

Establishment and Evaluation of Rat Model of Acute Coronary Syndrome with Combined Blood Stasis and Poison / 中国实验方剂学杂志

ObjectiveTo compare the feasibility of establishing the rat model of acute coronary syndrome with combined blood stasis and poison by lipopolysacharide （LPS） injection， ligation of coronary artery and different combinations of the two methods. MethodA total of 225 male SD rats were randomly divided into sham operation group， simple coronary artery ligation group， first injected LPS group ［LPS（5 mg·kg）injection 24 h before coronary artery ligation］ and follow injected LPS group ［LPS（5 mg·kg）injection 10 min after coronary artery ligation］. The indexes of each group were detected at 3， 24， 72 h after modeling， and the model was comprehensively evaluated. The general state and macroscopic evaluation indexes of traditional Chinese medicine （TCM） syndrome （tongue and pulse） of rats in each group were observed. ECG and echocardiography were used to evaluate cardiac function， and the myocardial ischemia and infarction areas were measured by Evans blue/2，3，5-triphenyltetrazolium chloride （TTC） staining. The content of creatine kinase isoenzyme （CK-MB）， lactate dehydrogenase （LDH）， creatine kinase （CK）， and troponin T （cTnT） in serum as well as interleukin-1 β （IL-1β） and IL-6 changes were determined by biochemical method or enzyme-linked immunosorbent assay （ELISA）. Hematology analyzer was adopted to determine the white blood cell （WBC） count， and the four coagulation indexes， platelet aggregation rate， hemorheology and other coagulation evaluation indexes were also detected. The myocardial tissue was observed by hematoxylin-eosin（HE）staining. ResultAfter 3 h of modeling， compared with the conditions in sham operation group， the R， G and B values of tongue of rats （P<0.01）， pulse amplitude （P<0.01）， and cardiac function in simple coronary artery ligation group were decreased， and the color of hypoglossal veins became purple（P<0.01）. The content of CK， LDH， cTnT， IL-1β and IL-6 in serum（P<0.05）， myocardial infarction area（P<0.01）， and total number of WBCs （P<0.05）were increased. Compared with simple coronary artery ligation group， first injected LPS group and follow injected LPS group had increased hypoglossal veins， decreased R value of tongue and elevated cTnT content （P<0.01）， while follow injected LPS group had reduced B value of tongue， decreased cardiac output （CO）（P<0.05）， increased IL-1β content， and thinned left ventricular anterior walls at end-systole （LVAWs）（P<0.01）. After 24 h of modeling， compared with sham operation group， simple coronary artery ligation group presented significantly decreased R， G and B values of tongue， lengthened purplish dark hypoglossal veins （P<0.01）， reduced pulse amplitude（P<0.01） and cardiac function， enlarged myocardial infarction area（P<0.01）， increased whole blood viscosity， platelet aggregation rate， fibrinogen （FIB）， shortened prothrombin time （PT） and thrombin time （TT）（P<0.01）， and elevated total number of WBCs （P<0.01）and content of CK， LDH， cTnT and IL-6 in serum（P<0.05）. Compared with the conditions in simple coronary artery ligation group， the pulse amplitude， R， G and B values of tongue （P<0.01）， and ejection fraction （EF） and fractional shortening （FS） scores （P<0.05）dropped， and hypoglossal veins were deepened and lengthened（P<0.05）， and cTnT content was increased（P<0.01）in first injected LPS group and follow injected LPS group. However， follow injected LPS group had thinned LVPWs， increased LDH content， platelet aggregation rate（P<0.05）， myocardial infarction area， and total number of WBC， level of IL-1β（P<0.05）， and shortened TT（P<0.01）. Additionally， 72 h after modeling， compared with sham operation group， simple coronary artery ligation group showed significantly reduced pulse amplitude， lowered R， G and B values of tongue， thickened and lengthened hypoglossal veins（P<0.01）， decreased cardiac function， and increased content of cTnT， FIB， whole blood viscosity（P<0.01），platelet aggregation rate， level of IL-6 and IL-1β（P<0.05）. Compared with the conditions in simple coronary artery ligation group， the hypoglossal veins of the first injected LPS group and the follow injected LPS group were more purple， and the content of cTnT was boosted（P<0.01）， whereas follow injected LPS group had decreased pulse amplitude， R， G and B values of tongue， EF and FS scores （P<0.05）， and enlarged myocardial infarction area（P<0.01）. ConclusionCompared with the other modeling methods and models at different modeling time， the established model by LPS injection 10 min after coronary artery ligation for 24 h was more consistent with the clinical characteristics of acute coronary syndrome with combined blood stasis and poison.

Establishment and Evaluation of Rat Model of Acute Coronary Syndrome with Combined Blood Stasis and Poison / 中国实验方剂学杂志

ObjectiveTo compare the feasibility of establishing the rat model of acute coronary syndrome with combined blood stasis and poison by lipopolysacharide （LPS） injection， ligation of coronary artery and different combinations of the two methods. MethodA total of 225 male SD rats were randomly divided into sham operation group， simple coronary artery ligation group， first injected LPS group ［LPS（5 mg·kg）injection 24 h before coronary artery ligation］ and follow injected LPS group ［LPS（5 mg·kg）injection 10 min after coronary artery ligation］. The indexes of each group were detected at 3， 24， 72 h after modeling， and the model was comprehensively evaluated. The general state and macroscopic evaluation indexes of traditional Chinese medicine （TCM） syndrome （tongue and pulse） of rats in each group were observed. ECG and echocardiography were used to evaluate cardiac function， and the myocardial ischemia and infarction areas were measured by Evans blue/2，3，5-triphenyltetrazolium chloride （TTC） staining. The content of creatine kinase isoenzyme （CK-MB）， lactate dehydrogenase （LDH）， creatine kinase （CK）， and troponin T （cTnT） in serum as well as interleukin-1 β （IL-1β） and IL-6 changes were determined by biochemical method or enzyme-linked immunosorbent assay （ELISA）. Hematology analyzer was adopted to determine the white blood cell （WBC） count， and the four coagulation indexes， platelet aggregation rate， hemorheology and other coagulation evaluation indexes were also detected. The myocardial tissue was observed by hematoxylin-eosin（HE）staining. ResultAfter 3 h of modeling， compared with the conditions in sham operation group， the R， G and B values of tongue of rats （P<0.01）， pulse amplitude （P<0.01）， and cardiac function in simple coronary artery ligation group were decreased， and the color of hypoglossal veins became purple（P<0.01）. The content of CK， LDH， cTnT， IL-1β and IL-6 in serum（P<0.05）， myocardial infarction area（P<0.01）， and total number of WBCs （P<0.05）were increased. Compared with simple coronary artery ligation group， first injected LPS group and follow injected LPS group had increased hypoglossal veins， decreased R value of tongue and elevated cTnT content （P<0.01）， while follow injected LPS group had reduced B value of tongue， decreased cardiac output （CO）（P<0.05）， increased IL-1β content， and thinned left ventricular anterior walls at end-systole （LVAWs）（P<0.01）. After 24 h of modeling， compared with sham operation group， simple coronary artery ligation group presented significantly decreased R， G and B values of tongue， lengthened purplish dark hypoglossal veins （P<0.01）， reduced pulse amplitude（P<0.01） and cardiac function， enlarged myocardial infarction area（P<0.01）， increased whole blood viscosity， platelet aggregation rate， fibrinogen （FIB）， shortened prothrombin time （PT） and thrombin time （TT）（P<0.01）， and elevated total number of WBCs （P<0.01）and content of CK， LDH， cTnT and IL-6 in serum（P<0.05）. Compared with the conditions in simple coronary artery ligation group， the pulse amplitude， R， G and B values of tongue （P<0.01）， and ejection fraction （EF） and fractional shortening （FS） scores （P<0.05）dropped， and hypoglossal veins were deepened and lengthened（P<0.05）， and cTnT content was increased（P<0.01）in first injected LPS group and follow injected LPS group. However， follow injected LPS group had thinned LVPWs， increased LDH content， platelet aggregation rate（P<0.05）， myocardial infarction area， and total number of WBC， level of IL-1β（P<0.05）， and shortened TT（P<0.01）. Additionally， 72 h after modeling， compared with sham operation group， simple coronary artery ligation group showed significantly reduced pulse amplitude， lowered R， G and B values of tongue， thickened and lengthened hypoglossal veins（P<0.01）， decreased cardiac function， and increased content of cTnT， FIB， whole blood viscosity（P<0.01），platelet aggregation rate， level of IL-6 and IL-1β（P<0.05）. Compared with the conditions in simple coronary artery ligation group， the hypoglossal veins of the first injected LPS group and the follow injected LPS group were more purple， and the content of cTnT was boosted（P<0.01）， whereas follow injected LPS group had decreased pulse amplitude， R， G and B values of tongue， EF and FS scores （P<0.05）， and enlarged myocardial infarction area（P<0.01）. ConclusionCompared with the other modeling methods and models at different modeling time， the established model by LPS injection 10 min after coronary artery ligation for 24 h was more consistent with the clinical characteristics of acute coronary syndrome with combined blood stasis and poison.

Identification and quality evaluation of germplasm resources of commercial Scutellaria baicalensis based on DNA barcode and HPLC / 中国中药杂志

Scutellaria baicalensis is a commonly used Chinese medicinal herb. In this study, we identified the germplasm resources of commercial S. baicalensis samples based on trnH-psbA, petA-psbJ, and ycf4-cemA sequences according to the available chloroplast genome sequencing results, and measured the content of baicalin by HPLC. Through the above means we determined the best DNA barcode that can be used to detect the germplasm resources and evaluate the quality of commercial S. baicalensis samples. A total of 104 samples were collected from 24 provinces, from which DNA was extracted for PCR amplification. The amplification efficiencies of trnH-psbA, petA-psbJ, and ycf4-cemA sequences were 100%, 59.62%, and 25.96%, respectively. The results of sequence analysis showed that 5, 4, and 2 haplotypes were identified based on trnH-psbA, petA-psbJ, and ycf4-cemA sequences, respectively. However, the sequences of haplotypes in commercial samples were different from that of the wild type, and the joint analysis of three fragments of S. baicalensis only identified 6 haplotypes. Furthermore, the phylogenetic analysis and genetic distance analysis indicated that trnH-psbA could be used to identify S. baicalensis from adulterants. The above analysis showed that trnH-psbA was the best fragment for identifying the germplasm resources of commercial S. baicalensis samples. We then analyzed the haplotypes(THap1-THap5) of commercial S. baicalensis samples based on trnH-psbA and found that THap2 was the main circulating haplotype of the commercial samples, accounting for 86.55% of the total samples, which indicated the scarce germplasm resources of commercial S. baicalensis samples. The content of baicalin in all the collected commercial S. baicalensis samples exceeded the standard in Chinese Pharmacopoeia and had significant differences(maximum of 12.21%) among samples, suggesting that the quality of commercial S. baicalensis samples varied considerably. However, there was no significant difference in baicalin content between different provinces or between different haplotypes. This study facilitates the establishment of the standard identification system for S. baicalensis, and can guide the commercial circulation and reasonable medication of S. baicalensis.

Effects of early life PM2.5 exposure on prefrontal cortex of offspring male rats / 中国应用生理学杂志

Objective: To investigate the effects of PM2.5 exposure at different stages of early life on the prefrontal cortex of offspring rats. Methods: Twelve pregnant SD rats were randomly divided into four groups: Control group (CG), Maternal pregnancy exposure group (MG), Early postnatal exposure group (EP) and Perinatal period exposure group (PP), 3 rats in each group. The pregnant and offspring rats were exposed to clean air or 8-fold concentrated PM2.5. MG was exposed from gestational day (GD) 1 to GD21. EP was exposed from postnatal day (PND) 1 to PND21, and PP was exposed from GD1 to PND21. After exposure, the prefrontal cortex of 6 offspring rats in each group was analyzed. HE staining was used to observe the pathological damage in the prefrontal cortex. ELISA was employed to detect neuroinflammatory factors, and HPLC/MSC was applied to determine neurotransmitter content. Western blot and colorimetry were applied for detecting astrocyte markers and oxidative stress markers, respectively. Results: Compared with MG and CG, the pathological changes of prefrontal cortex in PP and EP were more obvious. Compared with MG and CG, the neuroinflammatory factors (IL-1, IL-6, TNF-α) in PP and EP were increased significantly (P＜0.01), the level of MT were decreased significantly (P＜0.05), and the level of oxytocin (OT) showed a downward trend; the level of neurotransmitter ACh was also increased significantly (P＜0.01). Compared with MG and CG, the GFAP level of PP and EP showed an upward trend, the level of oxidative stress index SOD in PP and EP was decreased significantly (P＜0.01), and the level of ROS was increased significantly (P＜0.01). Compared with the offspring rats of CG and MG, the CAT level of PP was decreased significantly (P＜0.01, P＜0.05). Compared with the offspring rats of CG, the CAT level of EP was decreased significantly (P＜0.05). There was no significant difference in IL-1, IL-6, TNF-α, MT, OT, ACh, GFAP, SOD, ROS and CAT levels between PP and EP, or MG and CG. Conclusion: PM2.5 exposure in early life has adverse effects on the prefrontal cortex of offspring male rats, and early birth exposure may be more sensitive.

Effect of Buyang Huanwutang on Platelet Function and Inflammatory Cytokine in Rat Model of Acute Blood Stasis Induced by Multiple Factors / 中国实验方剂学杂志

ObjectiveTo explore the effect of Buyang Huanwutang （BYHWT） on platelet function and inflammatory cytokines in the rat model of acute blood stasis. MethodThe model of acute blood stasis was established with SD rats by ice water bath combined with injection of epinephrine. Rats were randomly assigned into four groups： normal group， model group， BYHWT （3.2 g·kg-1） group， and aspirin （60 mg·kg-1） group. The rats were injected with epinephrine hydrochloride on day 8 after 7 days of modeling. The macroscopic indexes of triditional Chinese medicine （TCM） syndrome including tongue manifestation and pulse manifestation were observed， while hemorheological indexes， blood coagulation， and platelet aggregation were detected. The serum levels of the inflammatory cytokine matrix metalloprotein-9 （MMP-9） and the adhesion factor intercellular adhesion molecule-1 （ICAM-1） and were determined by enzyme-linked immunosorbent assay （ELISA）. ResultThe pulse distention of rats in the model group was lower than that in the normal group （P<0.01）， while BYHWT improved the pulse distention of the rats with the syndrome of blood stasis （P<0.01）. In the model group， the tongue showed the characteristics of blood stasis syndrome， with dark purple veins at the tongue bottom and lower values of R， G， B on the tongue surface than those in the normal group （P<0.01）， which， however， can be recovered by BYHWT （P<0.01）. The blood viscosity at high， medium， and low shear stress and the plasma viscosity in the model group were higher than those in the normal group （P<0.01， P<0.05）. Compared with the model group， BYHWT restored the whole blood viscosity under high， medium and low shear stress and plasma viscosity （P<0.05，P<0.01）. The model group had shorter prothrombin time （PT）， shorter thrombin time （TT）， and higher fibrinogen （FIB） than the normal group （P<0.05， P<0.01）. BYHWT improved the TT and reduced the FIB in the rats with blood stasis syndrome （P<0.01）. The platelet aggregation rate induced by arachidonic acid （AA） and adenosine diphosphate （ADP） in the model group was higher than that in normal group （P<0.01） and BYHWT decreased the platelet aggregation rate of the rats with blood stasis syndrome （P<0.01）. The results of scanning electron microscopy showed that the model group exhibited excessive platelet activation， obvious pseudopodia， and increased aggregation of platelets compared with the normal group， while platelet activation and aggregation were rare in the BYHWT group. The serum levels of MMP-9 and ICAM-1 in the model group were higher than those in the normal group （P<0.01）， which were decreased in the BYHWT group （P<0.05， P<0.01）. ConclusionThe SD rats with the syndrome of acute blood stasis induced by ice water bath combined with injection of epinephrine demonstrate obvious changes in platelet function and morphology， inflammation， and abnormal cell adhesion. In the treatment of acute blood stasis in rats， BYHWT may reduce thrombosis and improve blood consistency and cohesion by mitigating inflammation， down-regulating cell adhesion factor overexpression， and improving platelet shape and function.

Cloning and subcellular location of the PnMYB1R1 gene from Panax notoginseng and expression analysis with different stresses / 药学学报

MYB transcription factors, one of the largest transcription factor families in plants, play an important role in signal transduction, plant growth and plant resistance. In this study a full-length cDNA of the PnMYB1R1 gene was cloned from Panax notoginseng. Sequence analysis, prokaryotic expression and purification, subcellular location, transcriptional activity analysis, tissue-specific analysis and expression analysis under different abiotic stresses was performed. The open reading frame (ORF) of PnMYB1R gene was 738 bp, encoding a protein of 245 amino acids with a predicted molecular mass (MW) of 27.0 kD. The sequence analysis and polygenetic analysis indicated that the PnMYB1R1 protein contains a conserved R3 domain, belonging to TRF-like protein in 1R-MYB-type transcription factors. The recombinant PnMYB1R1 protein was expressed in Escherichia coli BL21(DE3) cells using the prokaryotic expression vector pET28a-PnMYB1R1 and was purified. Subcellular localization analysis showed that PnMYB1R1 was localized in the nucleus. Transcriptional activity analysis indicated that the PnMYB1R1 transcription factor has transcriptional activation activity. Expression analysis indicated that PnMYB1R1 was primarily expressed in roots, followed by stems and leaves, and then rootlets. The expression level of PnMYB1R1 in root, stems, leaves and rootlets was influenced by salt, low temperature and drought treatment, while the abundance of PnMYB1R1 was significantly induced by salt stress in these tissues. These results provide valuable insights into the role of 1R-MYB transcription factors in plant defense.

Treatment options for colorectal cancer primary lesions with unresectable metastases at risk of acute abdomen / 国际肿瘤学杂志

For patients with metastatic colorectal cancer, it is still controversial whether the primary tumor needs surgery when the metastasis is unresectable. The effect of palliative surgical resection of the primary lesion on improving the prognosis and prolonging survival is still uncertain while the risks of acute abdomen trigger the discussion of early palliative surgical resection of the primary tumor. Evaluating and predicting the risk of acute abdomen complicated by colorectal cancer will help to choose the treatment of the primary lesion of unresectable metastatic colorectal cancer.

Research Progress on Molecular Mechanism of Lipid-lowering and Anti-inflammatory Effect of Zexietang / 中国实验方剂学杂志

Zexietang is a Chinese herbal compound prescription with a long history， which consists of Rhizoma alismatis and Atractylodes macrocephala. Zexietang comes from "Synopsis of Golden Chamber"， as "there is a drink under the heart， and its people are bitter and dizzy". Zexietang has the effect of removing water from drinking water and invigorating spleen for diuresis. Modern pharmacological studies have shown that its lipid-lowering and anti-inflammatory effects are very significant. It can be used to treat hyperlipidemia， anti-atherosclerosis and non-alcoholic fatty liver disease. With the continuous development of molecular biology， the research on the pharmacological effects of Zexietang， extracts and their monomers has deepened to the molecular level gradually， and the relevant mechanism of action has also been continuously elucidated. In terms of lipid-lowering effect of Zexietang， the levels of cytokines or receptors such as 3-hydroxy-3-methyl glutaryl coenzyme A reductase （HMGCR）， cholesterol 7α-hydroxylase （CYP7A1）， and liver X receptors （LXR） are affected. It is widely involved in adenosine 5'-monophosphate-activated protein kinase （AMPK） and peroxisome proliferators-activated receptors （PPAR） pathways， while Zexietang's anti-inflammatory effect mainly affects inflammatory factors such as interleukins （IL） and tumor necrosis factors （TNF）， and simultaneously nuclear factor κB （NF-κB）， toll-like receptors （TLR） and other cytokines or receptor-related pathways. In order to promote the further research and clinical application of Zexietang and contribute to the development of modernization of traditional Chinese medicine， the studies of the past 15 years on molecular mechanism of the lipid-lowering and anti-inflammatory effect of Zexietang， Alisma and Atractylodes extract as well as their monomer components were reviewed.

Effect of Shuangshen Ningxin Capsules on Cardiac Function and Hemodynamics in Rats with Coronary Microvascular Dysfunction / 中国实验方剂学杂志

Objective:To investigate the effect of Shuangshen Ningxin capsules （SSNX） on cardiac hemodynamics and cardiac function in rats with coronary microvascular dysfunction. Method:Rats were randomly divided into a sham operation group， a model group， a nicorandil group （5 mg·kg^-1）， and high- （180 mg·kg^-1）， medium- （90 mg·kg^-1）， and low-dose （45 mg·kg^-1） SSNX groups. Rats received corresponding drugs for 7 days. Two hours after the last administration， the model of coronary microvascular dysfunction was induced by left ventricular injection of embolic microspheres （40-120 μm， about 1 000 microspheres）. Twenty-four hours after modeling， left ventricular internal dimension in diastole （LVIDd）， left ventricular internal dimension in systole （LVIDs） left ventricular end-diastolic volume （LVEDV）， left ventricular end-systolic volume （LVESV）， stroke volume （SV）， cardiac output （CO）， left ventricular ejection fraction （EF）， and left ventricular shortening rate （FS） were detected by echocardiography. Cardiac catheterization was used to observe the arterial systolic blood pressure （SBP）， diastolic blood pressure （DBP）， left ventricular systolic pressure （LVSP）， left ventricular end-diastolic pressure （LVEDP）， maximum rate of increase in left ventricular pressure （LV+dp/dt_max）， and maximum rate of decrease in left ventricular pressure （LV-dp/dt_max）， and the mean arterial pressure （MAP） was calculated. Heart rate （HR） was calculated according to Ⅱ lead ECG. Biochemical analysis was carried out to detect the activities of creatine kinase （CK）， creatine kinase-MB （CK-MB）， and lactate dehydrogenase （LDH）. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum cardiac troponin T （cTnT）. Western blot was used to detect the protein expression of Caspase-3， Bcl-2， and Bax， and 2，3，5-triphenyltetrazolium chloride （TTC） staining to observe the area of myocardial infarction. Hematoxylin-eosin （HE） staining was used to observe the morphological changes of the myocardium. Result:As revealed by echocardiography， compared with the sham operation group， the model group showed reduced SV， CO， EF， and FS （<italic>P</italic><0.01）， and increased LVIDs and LVEDV （<italic>P</italic><0.01）. Compared with the model group， the SSNX groups showed increased EF （<italic>P</italic><0.05， <italic>P</italic><0.01） and FS （<italic>P</italic><0.01）， and the high- and medium-dose SSNX groups displayed reduced LVIDs and LVESV， and increased LVEDV， SV， and CO （<italic>P</italic><0.05， <italic>P</italic><0.01）. SBP， DBP， MAP， LVSP， LV+dp/dt_max， and LV-dp/dt_max in the model group were lower than those in the sham operation group （<italic>P</italic><0.01）， while there was no significant difference in HR. SSNX improved hemodynamics of rats， and increased SBP， DBP， MAP， LVSP， LV+dp/dt_max， LV-dp/dt_max， and HR as compared with the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. The serum CK， LDH， CK-MB， and cTnT levels in the model group were higher than those in the sham operation group （<italic>P</italic><0.01）. Compared with the model group， SSNX groups reduced serum CK， LDH， CK-MB， and cTnT （<italic>P</italic><0.05，<italic> P</italic><0.01）. Compared with the sham operation group， the model group displayed increased expression of Caspase-3 protein in the myocardium （<italic>P</italic><0.01） and reduced expression of Bcl-2 protein （<italic>P</italic><0.05）. The expression of Caspase-3 protein in the myocardium of SSNX groups was lower than that in the model group， and statistical difference was observed between the low-dose SSNX group and the model group （<italic>P</italic><0.05）. Compared with the model group， the SSNX groups exhibited increased expression of Bcl-2 in the rat myocardium， and the statistical difference was observed in the high-dose SSNX group <italic>（P</italic><0.01）. As demonstrated by the TTC staining， compared with the model group， SSNX groups showed reduced areas of myocardial infarction （<italic>P</italic><0.01）. The HE staining indicated that the pathological injury in myocardial tissues of the SSNX groups was relieved as compared with that in the model group. Conclusion:SSNX can significantly enhance the cardiac function after coronary microvascular dysfunction caused by embolic microspheres， improve cardiac hemodynamics， reduce the area of myocardial infarction， and decrease CK， LDH， CK-MB， and cTnT levels. The mechanism may be related to the inhibition of cardiomyocyte apoptosis to protect the myocardium.

Effect of Shuangshen Xionglian Granule on Vasodilation and PI3K/Akt/eNOS Pathway in Hyperhomocysteinemia Chronic Kidney Disease Rats / 中国实验方剂学杂志

Objective:To study the effect of Shuangshen Xionglian （SSXL） granules on vasculopathy and phosphatidylinositol 3 kinase （PI3K）/serine threonine kinase （Akt）/nitrogen oxide synthase （eNOS） signal in hyperhomocysteinemia chronic kidney disease rats. Method:Rats were randomly divided into 5 groups： sham operation group， model group， and high， medium and low-dose （8， 4， 2 g·kg-1） SSXL groups. The model of hyperhomocysteinemia chronic kidney disease in rats was established with high methionine feed combined with 5/6 nephrectomy. After 5/6 nephrectomy， continuous intragastric administration lasted for four weeks. Arterial blood pressure was measured at the 4th and 8th weeks after operation. At the end of the 8th week after the operation， blood was collected to determine serum creatinine， urea nitrogen， homocysteine （Hcy）， methionine and blood lipid. Western blot was used to detect the expressions of PI3K/Akt/eNOS pathway-related proteins， such as p-p85， p-Akt and p-Ser177 in thoracic aorta， and serum NO and eNOS were measured. The changes of endothelium-dependent relaxation and non-endothelium-dependent relaxation were measured by the method of isolated thoracic aorta ring. Pathological htoxylin eosin （HE） staining was used to observe the changes of renal tissue and thoracic aorta. Result:At the 8th week of the experiment， compared with the sham operation group， arterial systolic blood pressure， serum urea nitrogen， creatinine， Hcy， methionine， total cholesterol and low-density lipoprotein of the model group were significantly increased. Four weeks later after administration， arterial systolic blood pressure， serum urea nitrogen， Hcy， methionine， serum total cholesterol and serum low-density lipoprotein were significantly reduced in each dose group （P<0.05， P<0.01）. The creatinine in the SSXL 8， 4 g·kg-1 group was significantly reduced （P<0.05）. The nitric oxide content of SSXL in each dose groups were increased compared with that in the model group （P<0.05， P<0.01）， and the serum eNOS activity of the SSXL in the SSXL 8 g·kg-1 group was significantly increased compared with that in the model group （P<0.05）. The endothelium dependent and non-endothelium dependent vasodilation of thoracic aortic rings in the model group were significantly damaged. The cumulative concentration of acetylcholine （1×10-5.5~1×10-4 mmo1·L-1） in the SSXL 8 g·kg-1 group was significantly improved （P<0.05， P<0.01）. The diastolic degree of the vascular ring in the SSXL 8 g·kg-1 group was significantly higher than that in the model group （P<0.05）. Western blot results showed that the expressions of p-85， p-Akt and p-Ser177 in blood vessels increased in the sham group compared with those in the model group （P<0.01）. Compared with the model group， the phosphorylation level of this pathway was increased in the SSXL groups， and the expressions of p-Akt and p-Ser177 in the SSXL 8 g·kg-1 group were significantly increased （P<0.05）. The pathological results showed that the pathological changes of thoracic aorta and renal tissue in the dosages of SSXL were significantly reduced compared with those in the model group. Conclusion:SSXL granules can improve hyperhomocysteine and dyslipidemia in rats of chronic kidney disease with hyperhomocysteine， reduce serum creatinine， urea nitrogen levels and arterial systolic blood pressure， and improve vascular morphology and diastolic function， which may be related to the regulation of the PI3K/Akt/eNOS signaling pathway.

Application of optical trocar insertion in laparoscopic surgery after previous abdominal surgery / 临床肝胆病杂志

Objective To investigate the value of optical trocar insertion technique in establishing pneumoperitoneum in patients undergoing laparoscopic surgery after previous abdominal surgery. Methods A total of 29 patients, with a history of abdominal surgery, who planned to undergo laparoscopic liver surgery were enrolled and randomly divided into optical trocar insertion group and open approach group. The independent samples t -test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups; the Fisher's exact test was used for comparison of categorical data between groups; the Mann-Whitney U test was used for comparison of ranked data between groups. Results There were no procedure-related complications in either group. Compared with the open approach group, the optical trocar insertion group had a significantly shorter time required to establish pneumoperitoneum [35.00 (21.00-46.00) seconds vs 180.00 (152.50-252.50) seconds, U =0, P < 0.001] and a significantly smaller incision length [1.10(1.00-1.20) cm vs 2.80(2.45-3.00) cm, U =0, P < 0.001]. Conclusion Both optical trocar insertion and open approach for establishing pneumoperitoneum is relatively safe in patients undergoing laparoscopic liver surgery after previous abdominal surgery, while optical trocar insertion has the advantages of high efficiency and minimal invasiveness in establishing pneumoperitoneum.

Application of optical trocar insertion in laparoscopic surgery after previous abdominal surgery / 临床肝胆病杂志

Objective To investigate the value of optical trocar insertion technique in establishing pneumoperitoneum in patients undergoing laparoscopic surgery after previous abdominal surgery. Methods A total of 29 patients, with a history of abdominal surgery, who planned to undergo laparoscopic liver surgery were enrolled and randomly divided into optical trocar insertion group and open approach group. The independent samples t -test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups; the Fisher's exact test was used for comparison of categorical data between groups; the Mann-Whitney U test was used for comparison of ranked data between groups. Results There were no procedure-related complications in either group. Compared with the open approach group, the optical trocar insertion group had a significantly shorter time required to establish pneumoperitoneum [35.00 (21.00-46.00) seconds vs 180.00 (152.50-252.50) seconds, U =0, P < 0.001] and a significantly smaller incision length [1.10(1.00-1.20) cm vs 2.80(2.45-3.00) cm, U =0, P < 0.001]. Conclusion Both optical trocar insertion and open approach for establishing pneumoperitoneum is relatively safe in patients undergoing laparoscopic liver surgery after previous abdominal surgery, while optical trocar insertion has the advantages of high efficiency and minimal invasiveness in establishing pneumoperitoneum.

Clinical features related to lymphatic metastasis in grade 3 endometroid endometrial cancer: a retrospective cross-sectional study / 中华医学杂志(英文版)

BACKGROUND@#Endometrial cancer (EC) has been one of the most general cancers with respect to gynecological malignancies; however, there are debates on clinical strategies concerning treatments especially for patients with grade 3 (G3) endometroid endometrial cancer (EEC). Present study aimed to evaluate the lymphatic metastasis (LM) related factors and figure out the necessity of lymphadenectomy for G3 EEC patients.@*METHODS@#From January 2009 to April 2019, 3751 EC patients were admitted to Obstetrics and Gynecology Hospital of Fudan University. Clinical characteristics include age, grade, stage, and clinical pathological features. A total of 1235 EEC patients were involved in the multivariable analysis. Three hundred and eighty-one patients were involved in the survival analysis and the data attributed to sufficient follow-up information. Kaplan-Meier curve and log-rank test were utilized to analyze the survival rate.@*RESULTS@#Among the 1235 EEC patients, 181 (14.7%) were categorized as G3 and 1054 (85.3%) were grade 1 to grade 2 (G1-2). Multivariate analysis demonstrated that lymphovascular space invasion, adnexal involvement, and cervical stroma involvement were independent risk factors of LM in G3 cohort with odds ratio 3.4, 5.8, and 8.9; 95% confidence interval 1.1-10.6, 1.5-22.4, and 2.8-28.0, respectively. LM rates increased from 3.3% (3/92) to 75% (9/12) for G3 EEC cohort as related factor numbers increased from one to three. There were no differences between G3 and G1-2 EEC in overall survival and progression free survival. Additionally, no survival advantage was observed for G3 EEC patients at early stage with different plans of adjuvant treatment.@*CONCLUSIONS@#For G3 EEC patients without other pathological positive factor, the LM rate is lower than those with other pathological positive factor. Survival analysis showed no difference between G3 cohort and G1-2 cohort. Also, different adjuvant treatments had no impact on the overall survival for G3 EEC patients.

Difference of musculoskeletal ultrasound imaging of focus of knee joint tendon between patients with knee osteoarthritis and healthy subjects / 中国针灸

OBJECTIVE@#To observe the imaging features of focus of knee joint tendon in patients with knee osteoarthritis (KOA) by musculoskeletal ultrasound (MSUS) technique.@*METHODS@#One hundred KOA patients and 100 healthy subjects were included. All the KOA patients were palpated by the sequence of foot @*RESULTS@#The top-5 focus of knee tendon of KOA patients were located in medial inferior patella, medial tibial condyle, inferior patella, Zusanlici and Hedingci. The thickness of ligaments and tendons in extension and flexion positions in KOA patients were thicker than that in healthy subjects (@*CONCLUSION@#The focus of knee joint tendon in KOA patients shows significantly thickened musculoskeletal imaging features.

Characteristics of progestin-insensitive early stage endometrial cancer and atypical hyperplasia patients receiving second-line fertility-sparing treatment / 부인종양

Objective@#This study investigated the characteristics of progestin-insensitive endometrioid endometrial cancer (EEC) and atypical endometrial hyperplasia (AEH) patients receiving fertility-sparing treatments and assessed the therapeutic effects of second-line fertility-preserving treatments. @*Methods@#Three hundred and thirty-eight patients with EEC (n=75) or AEH (n=263) receiving fertility-preserving treatment were retrospectively analyzed. ‘Progestin-insensitive’ was defined as meeting one of the following criteria: 1) presented with progressed disease at any time during conservative treatment, 2) remained with stable disease after 7 months of treatment, and/or 3) did not achieve complete response (CR) after 10 months of treatment. Clinical characteristics and treatment results of progestin-insensitive patients receiving second-line treatment and those of progestin-sensitive patients were compared. @*Results@#Eight-two patients (59 AEH and 23 EEC) were defined as progestin-insensitive and 256 as progestin-sensitive. In multivariate analysis, body mass index ≥28.0 kg/m2 (odds ratio [OR]=1.898) and lesion size >2 cm (OR=2.077) were independent predictors of progestin-insensitive status. Compared to AEH patients, progestin-insensitive EEC patients had poorer second-line treatment responses (28-week cumulative CR rate after changing second-line treatment, 56.3% vs. 85.4%, p=0.011). No statistical difference was found in CR rate among different second-line treatments. @*Conclusion@#Obesity and larger lesion size were independent risk factors associated with progestin-insensitive status. In progestin-insensitive patients receiving second-line treatment, EEC patients had lower CR rate comparing with AEH patients. Further study with larger sample size is needed to evaluate efficacy of different second-line treatments for progestin insensitive patients.

Effect of Qihong Capsule on Heart Hemodynamics and Myocardial Consumption of Oxygen in Heart Failure Model Beagle / 中国实验方剂学杂志

Objective:To investigate the therapeutic effect of Qihong capsule on pentobarbital sodium induced heart failure in beagle dogs. Method:Thirty healthy adult beagle dogs were randomly divided into 6 groups, 6 in each group. They were normal group, model group, digoxin tablet group (40 μg·kg-1), Qihong capsule high, medium and low dose groups (2.6，1.3，0.65 g·kg-1). The heart failure model of beagle dogs was established by intravenous infusion of 2% pentobarbital sodium. The success standard of the model was that the maximum rate of rise of left ventricular pressure was reduced by 70%.The corresponding drugs were given through duodenum. The Ⅱ lead electrocardiogram, coronary blood flow, cardiac output, left ventricular pressure and maximum rate of left ventricular pressure rise were measured by multi-channel physiological recorder. The arterial oxygen content and coronary sinus oxygen content were measured by Roche blood oxygen analyzer at different time points, and the myocardial oxygen utilization rate was calculated. Result:After intravenous infusion of 2% pentobarbital sodium for about 15 minutes, beagle dogs began to show obvious symptoms of heart failure. The main manifestations were the increase of PR interval of Ⅱ lead electrocardiogram, the decrease of coronary blood flow, left ventricular pressure, cardiac output, cardiac output, venous oxygen content, and the increase of myocardial oxygen utilization rate (P<0.01) compared with the model group, Qihong capsule significantly increased coronary blood flow at 60-120 min after treatment (P<0.05). The cardiac output of 2.6 g·kg-1 Qihong capsule increased significantly at 45-60 min after treatment, with the maximum increase of about 16%, which was significantly different from that of model group (P<0.05). At the same time, it can increase the oxygen content of coronary sinus blood, which indicates that the myocardial oxygen supply is increased and the oxygen utilization rate is decreased. Qihong capsule 1.3 g·kg-1 group significantly increased the maximum rate of left ventricular pressure rise (P<0.05), the maximum increase rate was about 42%. Conclusion:Qihong capsule can increase coronary blood flow and venous blood oxygen content at the same time, make myocardial nutrient supply sufficient, reduce oxygen utilization rate, on this basis, Qihong capsule can further increase cardiac output and improve cardiac function, so as to play a protective role in heart failure.

Effect of Tianlong Tongxin Tablet on Acute Myocardial Ischemia Injury in Dogs / 中国实验方剂学杂志

Objective:To observe the effect of oral administration of Tianlong Tongxin tablet on acute myocardial ischemia and related indexes in experimental dogs. Method:The model of acute myocardial ischemia in dogs was established and the dogs were divided into the control group （equal amount of normo-cyclodintrin 10 g·kg-1）， Hexinshuang group （5 mg·kg-1）， Tianlong Tongxin tablet high， medium and low dose groups （1， 0.5， 0.25 g·kg-1） and the compound Danshen tablet group （0.144 g·kg-1）. Myocardial ischemia degree was measured by epicardium electrocardiogram， the range of myocardial infarction was determined by quantitative histology （N-BT staining）， and coronary blood flow， cardiac output， myocardial oxygen consumption， coronary resistance and peripheral resistance were measured. Meanwhile， serum creatine kinase （CK）， lactate dehydrogenase （LDH） and serum superoxide dismutase （SOD） activity and malondialdehyde （MDA） content were detected by optical kit. Result:As compared with the control group， Tianlong Tongxin tablet can reduce the myocardial ischemia degree （∑-ST） measured by the electrocardiogram of the pericardium （P<0.05）， reduce the infarcted area shown by N-BT staining （P<0.05）， reduce the venous oxygen content （P<0.05）， increase the coronary flow， cardiac output and myocardial oxygen consumption of anesthetized dogs， and reduce the coronary artery resistance and peripheral resistance （P<0.05）. However， there was no significant difference in the influence of serum CK， LDH， SOD activity and MDA content in serum. Conclusion:Tianlong Tongxin tablet can improve acute myocardial ischemia and myocardial infarction in dogs.