Effect of interleukin-6 on nucleated erythrocytes in lipopolysaccharide-induced preeclampsia rats / 解剖学报

Objective To explore the effect of interleukin (IL)-6 on nucleated erythrocytes in lipopolysaccharide (LPS)-induced preeclampsia rats. Methods ELISA and immunohistochemistry were used to detect the IL-6 in peripheral blood and placenta of preeclampsia and normal pregnancy; Flow cytometry and immunofluorescence were used to detect the maternal nucleated erythrocytes. Pregnant SD rats were randomly divided into 3 groups: the control, LPS and LPS +anti-IL-6 group; IL-6, the proportion of nucleated erythrocytes, JAK2/MEK and PI3K/Akt signal-related genes were detected. Results The IL-6 of preeclampsia was higher than that of normal patients. Compared with the Control group, IL-6, the proportion of nucleated erythrocytes and JAK2, P85, Akt, P65, IL-IB mRNA of LPS group increased, the fetal weight decreased; Compared with the LPS group, IL-6, the proportion of nucleated erythrocytes and JAK2, P85, Akt, P65 and IL-IB mRNA of the LPS + anti-IL-6 group decreased. Conclusion The up-regulation of IL-6 of preeclampsia patients is accompanied by increased nucleated erythrocytes in peripheral blood. Neutralizing IL-6 in vivo may down-regulate JAK2/ PI3K/Akt/NF-KB-signal-mediated IL-IB to protect preeclampsia rats.

Treatment of intrauterine adhesions in rats with hypoxia-cultured BMSC-derived exosomes / 中华妇产科杂志

Objective: To perform intrauterine adhesion modeling, and to investigate the repair effect of hypoxic treated bone marrow mesenchymal stem cells (BMSC) and their derived exosomes (BMSC-exo) on endometrial injury. Methods: BMSC and their exosomes BMSC-exo extracted from rats' femur were cultured under conventional oxygen condition (21%O2) or hypoxia condition (1%O2). Intrauterine adhesion modeling was performed on 40 healthy female SD rats by intrauterine injection of bacterial lipopolysaccharide after curettage. On the 28th day of modeling, 40 rat models were randomly divided into five groups, and interventions were performed: (1) NC group: 0.2 ml phosphate buffered solution was injected into each uterine cavity; (2) BMSC group: 0.2 ml BMSC (1×106/ml) with conventional oxygen culture was injected intrauterine; (3) L-BMSC group: 0.2 ml of hypoxic cultured BMSC (1×106/ml) was injected intrauterine; (4) BMSC-exo group: 0.2 ml of BMSC-exo cultured with conventional oxygen at a concentration of 500 μg/ml was injected into the uterine cavity; (5) L-BMSC-exo group: 0.2 ml hypoxic cultured BMSC-exo (500 μg/ml) was injected intrauterine. On the 14th and 28th day of treatment, four rats in each group were sacrificed by cervical dislocation after anesthesia, and endometrial tissues were collected. Then HE and Masson staining were used to observe and calculate the number of glands and fibrosis area in the endometrium. The expressions of angiogenesis related cytokines [vascular endothelial growth factor A (VEGFA) and CD31], and fibrosis-related proteins [collagen-Ⅰ, collagen-Ⅲ, smooth muscle actin α (α-SMA), and transforming growth factor β1 (TGF-β1)] in endometrial tissues were detected by western blot. Results: (1) HE and Masson staining showed that the number of endometrial glands in L-BMSC group, BMSC-exo group and L-BMSC-exo group increased and the fibrosis area decreased compared with NC group on the 14th and 28th day of treatment (all P<0.05). Noteworthily, the changes of L-BMSC-exo group were more significant than those of BMSC-exo group (all P<0.05), and the changes of BMSC-exo group were greater than those of BMSC group (all P<0.05). (2) Western blot analysis showed that, compared with NC group, the expressions of collagen-Ⅲ and TGF-β1 in BMSC group, L-BMSC group, BMSC-exo group and L-BMSC-exo group decreased on the 14th and 28th day of treatment (all P<0.05). As the treatment time went on, the expressions of fibrosis-related proteins were different. Compared with BMSC group, the expressions of collagen-Ⅲ, α-SMA and TGF-β1 in the BMSC-exo group and L-BMSC group decreased on the 28th day (all P<0.05). Moreover, the expressions of collagen-Ⅲ and TGF-β1 in L-BMSC-exo group were lower than those in BMSC-exo group on the 28th day (all P<0.05). And the expressions of collagen-Ⅰ, α-SMA and TGF-β1 in L-BMSC-exo group were lower than those in L-BMSC group on the 28th day (all P<0.05). (3) The results of western blot analysis of VEGFA and CD31 showed that, the expressions of VEGFA and CD31 in BMSC group, L-BMSC group, BMSC-exo group and L-BMSC-exo group increased on the 14th and 28th day of treatment compared with NC group (all P<0.05). Treatment for 28 days, the expressions of VEGFA and CD31 in BMSC-exo group and CD31 in L-BMSC group were higher than those in BMSC group (all P<0.05). Moreover, the expressions of VEGFA and CD31 in L-BMSC-exo group were higher than those in BMSC-exo group and L-BMSC group on the 28th day (all P<0.05). Conclusions: Treatment of BMSC and their exosomes BMSC-exo with hypoxia could promote endometrial gland hyperplasia, inhibit tissue fibrosis, and further repair the damaged endometrium in rats with intrauterine adhesion. Importantly, hypoxic treatment of BMSC-exo is the most effective in intrauterine adhesion rats.

Read-through circular RNA rt-circ-HS promotes hypoxia inducible factor 1α expression and renal carcinoma cell proliferation, migration and invasiveness / 北京大学学报(医学版)

OBJECTIVE@#To identify and characterize read-through RNAs and read-through circular RNAs (rt-circ-HS) derived from transcriptional read-through hypoxia inducible factor 1α (HIF1α) and small nuclear RNA activating complex polypeptide 1 (SNAPC1) the two adjacent genes located on chromosome 14q23, in renal carcinoma cells and renal carcinoma tissues, and to study the effects of rt-circ-HS on biological behavior of renal carcinoma cells and on regulation of HIF1α.@*METHODS@#Reverse transcription-polymerase chain reaction (RT-PCR) and Sanger sequencing were used to examine expression of read-through RNAs HIF1α-SNAPC1 and rt-circ-HS in different tumor cells. Tissue microarrays of 437 different types of renal cell carcinoma (RCC) were constructed, and chromogenic in situ hybridization (ISH) was used to investigate expression of rt-circ-HS in different RCC types. Small interference RNA (siRNA) and artificial overexpression plasmids were designed to examine the effects of rt-circ-HS on 786-O and A498 renal carcinoma cell proliferation, migration and invasiveness by cell counting kit 8 (CCK8), EdU incorporation and Transwell cell migration and invasion assays. RT-PCR and Western blot were used to exa-mine expression of HIF1α and SNAPC1 RNA and proteins after interference of rt-circ-HS with siRNA, respectively. The binding of rt-circ-HS with microRNA 539 (miR-539), and miR-539 with HIF1α 3' untranslated region (3' UTR), and the effects of these interactions were investigated by dual luciferase reporter gene assays.@*RESULTS@#We discovered a novel 1 144 nt rt-circ-HS, which was derived from read-through RNA HIF1α-SNAPC1 and consisted of HIF1α exon 2-6 and SNAPC1 exon 2-4. Expression of rt-circ-HS was significantly upregulated in 786-O renal carcinoma cells. ISH showed that the overall positive expression rate of rt-circ-HS in RCC tissue samples was 67.5% (295/437), and the expression was different in different types of RCCs. Mechanistically, rt-circ-HS promoted renal carcinoma cell proliferation, migration and invasiveness by functioning as a competitive endogenous inhibitor of miR-539, which we found to be a potent post-transcriptional suppressor of HIF1α, thus promoting expression of HIF1α.@*CONCLUSION@#The novel rt-circ-HS is highly expressed in different types of RCCs and acts as a competitive endogenous inhibitor of miR-539 to promote expression of its parental gene HIF1α and thus the proliferation, migration and invasion of renal cancer cells.

Clinical characteristics of 111 cases with mucopolysaccharidosis ⅣA / 中华儿科杂志

Objective: To analyze the clinical characteristics of patients with Mucopolysaccharidosis ⅣA (MPS ⅣA). Methods: A retrospective study was conducted on 111 patients with MPS ⅣA in Xinhua Hospital of Shanghai Jiao Tong University School of Medcine from December 2008 to August 2020, confirmed by enzyme activity and genetic testing. General situation, clinical manifestations and enzyme activity test results were analyzed. According to the clinical manifestations, it can be divided into severe, intermediate and mild group. The independent sample t test was used to compare the birth body length and weight of children with that of normal boys and girls, and group comparisons of enzyme activities were evaluated by median test. Results: One hundred and eleven unrelated patients, 69 males and 42 females, were classified into 3 subtypes: severe (n=85), intermediate (n=14), and mild (n=12). The age at symptom onset were 1.6 (1.0, 3.0) years, and at diagnosis were 4.3 (2.8, 7.8) years. Skeletal manifestations were observed in all patients and consisted mainly of pectus carinatum (96/111, 86.5%), motor dysfunction (78/111, 70.3%), spinal deformity (71/111, 64.0%), growth retardation (64/111, 57.7%), joint laxity (63/111, 56.8%) and genu valgum (62/111, 55.9%). Eighty-eight patients (88/111, 79.3%) with MPS ⅣA were also along with non-skeletal manifestations, mainly including snoring (38/111, 34.2%), coarse faces (34/111, 30.6%), and visual impairment (26/111, 23.4%). The most common skeletal manifestation was pectus carinatum (79 cases), and non-skeletal manifestation was snoring (30 cases) and coarse faces (30 cases) in severe patients, pectus carinatum (13 cases) and snoring (5 cases) in intermediate type, motor dysfunction (11 cases) and snoring (3 cases) and visual impairment (3 cases) in mild patients. The height and weight of severe patients began to fall below -2 s at 2-<5 years and 5-<7 years, respectively. At the age of 10-<15 years, the standard deviation score of the height of severe patients reached (-6.2±1.6) s in males and (-6.4±1.2) s in females, and the score of weight got (-3.0±1.1) s in males and (-3.5±0.5) s in females. The height of intermediate patients began to fall below -2 s at the age of 7-<10 years, and the standard deviation score of height were -4.6 s and -3.6 s in 2 males, and -4.6 s and -3.8 s in 2 females at the age of 10-<15 years. The weight remained within -2 s in 72.0% (18/25) of intermediate patients compared to age-matched healthy children. In the mild patients with MPS ⅣA, the mean standard deviation score of height and weight was within -2 s. The enzyme activities of mild patients (2.02 (1.05, 8.20) nmol/(17 h·mg)) were both significantly higher than that of intermediate (0.57 (0.47, 0.94) nmol/(17 h·mg)) and severe (0.22 (0, 0.59) nmol/(17 h·mg)) patients (Z=9.91, 13.98, P=0.005, 0.001), and the enzyme activity of intermediate patients was significantly higher than that of severe patients (Z=8.56, P=0.010). Conclusions: The clinical manifestations of MPS ⅣA are charactered by pectus carinatum, motor function impairment, spinal deformity and growth retardation. The clinical characteristics, growth rate and enzyme activity differ among the 3 subtypes of MPS ⅣA.

Predictors of efficacy of corticosteroid switching from abiraterone plus prednisone to dexamethasone in patients with metastatic castration-resistant prostate cancer / 亚洲男科学杂志(英文版)

Corticosteroid switching can reverse abiraterone resistance in some patients with metastatic castration-resistant prostate cancer (mCRPC). Here, we investigated the potential biomarkers for predicting the efficacy of corticosteroid switching during treatment with abiraterone acetate (AA). We retrospectively analyzed 101 mCRPC patients receiving corticosteroid switching from West China Hospital and Sun Yat-Sen University Cancer Center between January 2016 and December 2018. All cases received AA plus prednisone as first-line therapy during mCRPC. Primary end points were biochemical progression-free survival (bPFS) and overall survival (OS). The risk groups were defined based on multivariate analysis. A total of 42 (41.6%) and 25 (24.8%) patients achieved 30% and 50% decline in prostate-specific antigen (PSA), respectively, after corticosteroid switching. The median bPFS and median OS on AA plus dexamethasone were 4.9 (95% confidence interval [CI]: 3.7-6.0) months and 18.8 (95% CI: 16.2-30.2) months, respectively. Aldo-keto reductase family 1 member C3 (AKR1C3) expression (hazard ratio [HR]: 2.15, 95% Cl: 1.22-3.80, P = 0.008) and baseline serum alkaline phosphatase (ALP; HR: 4.95, 95% Cl: 2.40-10.19, P < 0.001) were independent predictors of efficacy before corticosteroid switching in the multivariate analysis of bPFS. Only baseline serum ALP >160 IU l-1 (HR: 3.41, 95% Cl: 1.57-7.38, P = 0.002) together with PSA level at switch ≥50 ng ml-1 (HR: 2.59, 95% Cl: 1.22-5.47, P = 0.013) independently predicted poorer OS. Based on the predictive factors in multivariate analysis, we developed two risk stratification tools to select candidates for corticosteroid switching. Detection of serum ALP level, PSA level, and tissue AKR1C3 expression in mCRPC patients could help make clinical decisions for corticosteroid switching.

Analysis of lactate dehydrogenase gene polymorphisms and prediction of B cell epitopes in four human Plasmodium species / 中国血吸虫病防治杂志

Objective To analyze the polymorphism of Plasmodium lactate dehydrogenase (pLDH) gene and predict B-cell epitopes in pLDH peptides in four species of human malaria parasites. Methods The blood samples and epidemiological characteristics were collected from malaria cases in Yunnan Province registered in the National Notifiable Disease Report System. The pLDH genes of four human Plasmodium species were amplified using nested PCR assay and sequenced. The polymorphisms of pLDH genes was analyzed using the software MEGA version 7.0.26 and DnaSP version 5.10, and the B-cell epitopes were predicted in pLDH peptides using the Immune Epitope Database (IEDB). Results The sequences of P. vivax LDH (PvLDH), P. falciparum LDH (PfLDH), P. ovale LDH (PoLDH) and P. malariae LDH (PmLDH) genes were obtained from 153, 29, 17 and 11 blood samples from patients with P. vivax, P. falciparum, P. ovale and P. malariae malaria, respectively, which included 15, 2, 4 and 2 haplotypes and had a nucleotide diversity (π) of 0.104. A high level of intra-species differentiation was seen in the PoLDH gene (π = 0.012), and the π values were all < 0.001 for PvLDH, PfLDH and PmLDH genes. Active regions of B-cell antigen were predicted in the pLDH peptide chain of four human malaria parasites, of 4 to 5 in each chain, and the activity score was approximately 0.430. Among these peptide chains, the “86-PGKSDKEWNRD-96” short-peptide was a B-cell epitope shared by all four species of human malaria parasites, and the “266-GQYGHS (T)-271” short-peptide was present in PvLDH and PoLDH peptide chains, while “212-EEVEGIFDR-220” was only found in the PvLDH peptide chain, and “208-LISDAE-213” was only seen in the PfLDH peptide chain. Conclusions The PoLDH gene polymorphism may be derived from the weak negative purification selection, while PvLDH, PfLDH and PmLDH genes may maintain a relatively conservative state. There may be two B-cell epitopes “212-EEVEGIFDR-220” and “208-LISDAE-213” in the proximal region of the C terminal in the pLDH peptide chain, which is feasible to differentiate between P. vivax and P. falciparum infections.

Multidetector CT Characteristics of Fumarate HydrataseDeficient Renal Cell Carcinoma and Papillary Type II Renal Cell Carcinoma

Objective@#To investigate the multidetector computed tomography (MDCT) features of fumarate hydratase-deficient renal cell carcinoma (FH-deficient RCC) with germline or somatic mutations, and compare them with those of papillary type II RCC (pRCC type II). @*Materials and Methods@#A total of 24 patients (mean ± standard deviation, 40.4 ± 14.7 years) with pathologically confirmed FH-deficient RCC (15 with germline and 9 with somatic mutations) and 54 patients (58.6 ± 12.6 years) with pRCC type II were enrolled. The MDCT features were retrospectively reviewed and compared between the two entities and mutation subgroups, and were correlated with the clinicopathological findings. @*Results@#All the lesions were unilateral and single. Compared with pRCC type II, FH-deficient RCC was more prevalent among younger patients (40.4 ± 14.7 vs. 58.6 ± 12.6, p < 0.001) and tended to be larger (8.1 ± 4.1 vs. 5.4 ± 3.2, p = 0.002). Cystic solid patterns were more common in FH-deficient RCC (20/24 vs. 16/54, p < 0.001), with 16 of the 20 (80.0%) cystic solid tumors having showed typical polycystic and thin smooth walls and/or septa, with an eccentric solid component. Lymph node (16/24 vs. 16/54, p = 0.003) and distant (11/24 vs. 3/54, p < 0.001) metastases were more frequent in FH-deficient RCC. FHdeficient RCC and pRCC type II showed similar attenuation in the unenhanced phase. The attenuation in the corticomedullary phase (CMP) (76.3% ± 25.0% vs. 60.2 ± 23.6, p = 0.008) and nephrographic phase (NP) (87.7 ± 20.5, vs. 71.2 ± 23.9, p = 0.004), absolute enhancement in CMP (39.0 ± 24.8 vs. 27.1 ± 22.7, p = 0.001) and NP (50.5 ± 20.5 vs. 38.2 ± 21.9, p = 0.001), and relative enhancement ratio to the renal cortex in CMP (0.35 ± 0.26 vs. 0.24 ± 0.19, p = 0.001) and NP (0.43 ± 0.24 vs. 0.29 ± 0.19, p < 0.001) were significantly higher in FH-deficient RCC. No significant difference was found between the FH germline and somatic mutation subgroups in any of the parameters. @*Conclusion@#The MDCT features of FH-deficient RCC were different from those of pRCC type II, whereas there was no statistical difference between the germline and somatic mutation subgroups. A kidney mass with a cystic solid pattern and metastatic tendency, especially in young patients, should be considered for FH-deficient RCC.

Mechanism of Potentilla discolor in treating UC by regulating mitochondrial autophagy / 中国中药杂志

To evaluate the therapeutic effect of Potentilla discolor on 2,4,6-trinitrobenzensulfonic acid(TNBS)-induced experimental ulcerative colitis(UC) in rats and to determine its therapeutic mechanism through mitochondrial autophagy, immune cells, and cytokines. A rat model of UC was established by TNBS-ethanol enema. Rats were divided into six groups: control, UC model, sulfasalazine(positive drug), and high-dose, moderate-dose, and low-dose ethanol extract groups. After 14-day continuous administration of the corresponding drugs, the disease activity index(DAI) and hematoxylin and eosin(HE) were evaluated. The morphological structure of mitochondria was observed by using transmission electron microscope(TEM), mitophagy-related mRNA expression was detected by using Real-time quantitative polymerase chain reaction(qRT-PCR), immune cell differentiation in rat serum was detected by using flow cytometry(FCM), and cytokine expression in colon tissues of rats was detected by protein microarray. The results showed that compared with the model group, each dose group of P. discolor could significantly reduce the DAI of UC model rats, and decrease the degree of inflammatory cells infiltration in the colon tissue of UC model rats. Meanwhile the expressions of T cells and Th cells in the serum increased significantly, the expression of Tc cells in the serum decreased significantly. Transmission electron microscope found that there was fusion of mitochondria and lysosomes in the colon tissue of the administration group. The expressions of mitochondrial autophagy related genes NF-κB, p62 and parkin were significantly increased in colon tissues. The results of protein chip showed that compared with the model group, the high dose group of P. discolor could significantly regulate the expression of cytokines. In conclusion, these results suggested that P. discolor improved TNBS-induced acute ulcerative colitis in rats by regulating the mitochondrial autophagy and the inflammatory factor expression.

Analysis of Risk Factors for a Poor Prognosis in Patients with Anti-N-Methyl-D-Aspartate Receptor Encephalitis and Construction of a Prognostic Composite Score

Background@#and Purpose: Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is the most-common form of autoimmune encephalitis, but its early diagnosis is challenging.This study aimed to identify the risk factors for a poor prognosis in anti-NMDAR encephalitis and construct a prognostic composite score for obtaining earlier predictions of a poor prognosis. @*Methods@#We retrospectively analyzed the clinical data, laboratory indexes, imaging findings, and electroencephalogram (EEG) data of 60 patients with anti-NMDAR encephalitis. The modified Rankin Scale (mRS) scores of patients were collected when they were discharged from the hospital. The mRS scores were used to divide the patients into two groups, with mRS scores of 3–6 defined as a poor prognosis. Logistic regression analysis was used to analyze independent risk factors related to a poor prognosis. @*Results@#This study found that 23 (38.3%) and 37 (61.7%) patients had good and poor prognoses, respectively. Logistic regression analysis showed that age, disturbance of consciousness at admission, and ≥50% slow waves on the EEG were significantly associated with patient outcomes. An age, consciousness, and slow waves (ACS) composite score was constructed to predict the prognosis of patients with anti-NMDAR encephalitis at an early stage based on regression coefficients. @*Conclusions@#Age, disturbance of consciousness at admission, and ≥50% slow waves on the EEG were independent risk factors for a poor prognosis. The ACS prognostic composite score could play a role in facilitating early predictions of the prognosis of anti-NMDAR encephalitis.

Effect of Xiao Jianzhongtang on AMPK/PGC1-α Signal Pathway in Skeletal Muscle of Exercise Fatigue Mice / 中国实验方剂学杂志

Objective:To observe the effect of Xiao Jianzhongtang on Adenylate-activated protein kinase/peroxidase proliferation-activated receptor coactivator 1-α （AMPK/PGC1-α） signaling pathway in skeletal muscle of exercise fatigue mice.Method:Forty Kunming mice were randomly divided into normal group， model group， Buzhong Yiqitang group and Xiao Jianzhongtang group， with 10 mice in each group. The model group， Buzhong Yiqitang group and Xiao Jianzhongtang group were trained on the treadmill to establish a fatigue model， and the normal group did not apply any intervention. At the same time as the treadmill training， the model group was given the same amount of normal saline. Xiao Jianzhongtang was administered with 5 g·kg-1 of medicine， and Buzhong Yiqitang was administered with 2.8 g·kg-1 of medicine for 6 days. After the experiment， the weight of each group of mice and the time of running out of exhaustion were measured，the colorimetric method was used to detect the serum urea （UREA）， lactate dehydrogenase （LDH）， muscle glycogen （MG）， and skeletal muscle of each group of mice Na+-K+-ATPase， Ca2+-Mg2+-ATPase content， pathological changes of skeletal muscle of each group were observed by hematoxylin-eosin （HE） staining， Western blot was used to detect the protein expression of AMPK and PGC1-α in skeletal muscle of each group .Result:Compared with normal group， the body weight of model group significantly decreased （P<0.01）， and the contents of Na+-K+-ATPase， Ca2+-Mg2+-ATPase， LDH， and MG significantly decreased （P<0.05，P<0.01）. The content of UREA increased significantly （P<0.01）， and the expression of AMPK and PGC1-α protein increased significantly （P<0.01）. Compared with model group， the mice in the Xiao Jianzhongtang group had significantly increased body weight （P<0.05）， significantly increased the time spent on treadmill exhaustion（P<0.01）， Na+-K+-ATPase， Ca2+-Mg2+-ATPase， LDH， and MG. The content increased significantly（P<0.05， P<0.01）， the content of UREA decreased significantly （P<0.01）， and the expression of AMPK and PGC1-α protein increased significantly （P<0.01）.Conclusion:Xiao Jianzhongtang has an anti-exercise fatigue effect， which may be related to enhancing skeletal muscle AMPK/PGC1-α pathway，enhancing mitochondrial oxidative phosphorylation，reducing accumulation of metabolites，slowing down glycogen consumption and decomposition，and enhancing skeletal muscle energy synthesis.

Protective Effect of Wutou Chishizhi Wan on Vascular Endothelial Cells and Oxidative Stress in Myocardial Ischemia-reperfusion Rats / 中国实验方剂学杂志

Objective:To investigate the protective effect of Wutou Chishizhi Wan on myocardial ischemia reperfusion injury （MIRI） in rats， and observe its effect on such mechanisms as coagulation function， vascular endothelial cells and oxidative stress in rats. Method:A total of 40 SD rats were randomly divided into normal group， model group， positive drug group （Urokinase group） and Wutou Chishizhi Wan group， with 10 rats in each group. Except for the normal group， rat myocardial ischemia-reperfusion injury models were established. The changes of heart rate （HR） at 10 min before ischemia， 30 min after ischemia and 30， 60， 120 min （T0，T1，T2，T3，T4）， and the change of electrocardiogram （ECG） J point after modeling in rats were observed. The pathological changes of rat myocardial tissue were observed by hematoxylin-eosin （HE） staining. The changes of four indexes of coagulation ［prothrombin time （PT）， activated partial thromboplastin time （APTT）， thrombin time （TT）， fibrinogen content decreased significantly （FIB）］ in rats were observed. The contents of endothelin-1 （ET-1）， thromboxane A2 （TXA2） and prostacyclin （PGI2） in serum and myocardium levels of superoxide dismutase （SOD）， malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） of MIRI rats were observed. Western blot assay was used for the detection of oxidative stress protein Keap1 and transcription factor-E2-related factor （Nrf2） expression levels in rat myocardial tissue. Result:Compared with the normal group， the ECG of MIRI rats showed significant myocardial ischemic injury-like changes， ST segment was significantly elevated， J point was significantly increased， and the incidences of HR in T1， T2， T3 and T4 were significantly reduced （P<0.05， P<0.01）. Compared with the model group， Wutou Chishizhi Wan significantly reduced ECG J-point changes in MIRI rats， while increased the incidence of HR in T1， T2， T3 and T4 （P<0.05， P<0.01）. Compared with the normal group， PT， APTT and TT in the model group were significantly shortened （P<0.01）， FIB content was significantly increased （P<0.01）， and the serum PGI2 level decreased and TXA2 and ET-1 levels increased significantly in the model group （P<0.01）. SOD content and GSH-Px activities of myocardial tissue in the model group were significantly reduced （P<0.01）， whereas the MDA content was increased （P<0.01）. Compared with the model group， PT of the Wutou Chishizhi Wan group was prolonged （P<0.05） and APTT slightly prolonged， TT significantly prolonged （P<0.01）， FIB content decreased （P<0.05）， serum PGI2 increased （P<0.05）， TXA2 and ET-1 decreased significantly in the Wutou Chishizhi Wan group （P<0.01）， myocardial MDA content decreased， and SOD content and GSP-Px activity increased significantly （P<0.01）. Meanwhile， the Wutou Chishizhi Wan group was able to activate the Keap1/Nrf2 signaling pathway， which significantly increased Nrf2 expression and significantly decreased Keap1 expression （P<0.01）. Conclusion:Wutou Chishizhi Wan group can protect myocardial injury in MIRI rats. The specific mechanism is to protect MIRI by regulating vascular endothelial cell homeostasis and oxidative stress levels and activating Keap1/Nrf2 signaling pathway.

Expression of Krupple-like factor 6 and inducible nitric oxide synthase in apoptosis of RAW264.7 infected by pseudomonas aeruginosa supernatant / 中国药理学通报

Aim To detect the expression of Krupple-like factor 6 ( KLF6) and inducible nitric oxide syn-thase (iNOS) in mouse mononuclear macrophage leukemia cells infected with pseudomonas aeruginosa (PA) supernatant, and to investigate the role of KLF6 protein and iNOS protein in the apoptosis of RAW264. 7 infected with PA supernatant. Methods The effect of PA supernatant on the proliferation of RAW264. 7 was detected by MTT assay. The apoptotic rate was detected by flow cytometry. The morphological changes of RAW264.7 were detected by Hoechst 33342 staining. The effect of PA supernatant on KLF6 and iNOS protein expression was detected by Western blot. Results PA supernatant inhibited the proliferation of RAW264.7 in a concentration- A nd time-depend-ent manner. PA supernatant significantly increased the percentage of RAW264.7 apoptosis and promoted the expression of KLF6 and iNOS. Conclusions PA can inhibit the proliferation of RAW264. 7, which may induce apoptosis through KLF6 and iNOS protein expres-sion.

Stem cell repair of intrauterine adhesions: preliminary achievements and clinical translation / 中国组织工程研究

BACKGROUND: Stem cells exhibit immeasurable application potentials in tissue and organ repair, but stem cell transplantation for the treatment of intrauterine adhesions is still in the initial stage. OBJECTIVE: To review the research progress in the stem cell repair of intrauterine adhesions. METHODS: We retrieved ISI Web of Science database, PubMed database and CNKI database for representative clinical research, basic research and reviews concerning stem cell therapy for intrauterine adhesions. The keywords were "intrauterine adhesion, metrosynizesis, Asherman's syndrome, stem cell, endometrial stem cell, stem cell transplantation" in English and Chinese, respectively. After repetitive studies were excluded, 43 articles were reviewed in the result analysis. RESULTS AND CONCLUSION: Hysteroscopic transcervical resection of adhesions is an ideal treatment for intrauterine adhesions, but the postoperative recurrence rate of intrauterine adhesions is still high. The human endometrium has high proliferative activity, and the endometrium of a woman of childbearing age may experience growth, differentiation and exfoliation for over 400 times, indicating the existence of endometrial stem cells.Endometrial stem cells have been isolated from the endometrium,successfully cultured in vitro and induced for directional differentiation. However, studies on endometrial stem cell transplantation for intrauterine adhesions are still in its infancy. Basic research on stem cells will facilitate its application to clinical practice.

Effect of FAM83A on stem cell-like traits, chemosensitivity and radiosensitivity of pancreatic cancer cells PANC-1 / 中华放射医学与防护杂志

Objective To investigate the effect of FAM83A on the stem cell-like phenotype, chemosensitivity and radiosensitivity of PANC-1 cells, aiming to provide new ideas for clinical combination therapy of pancreatic cancer. Methods The PANC-1 cells with stable silencing FAM83A were constructed by using lentivirus and validated by qPCR and Western blot. Flow cytometry was used to detect the number of CD133 positive cells and cellular apoptosis; the sphere formation assay was used to test the ability of sphere formation of PANC-1 cells;the effect of gecitabine on the cell viability was detected by MTT assay;the effect of radiation on the proliferation of PANC-1 cells was detected by colony formation assay; the effect of FAM83A on Wnt/β-catenin pathway was examined by Western blot. Results The expressions of FAM83A protein ( 0.83 ± 0.08 ) and mRNA ( 0.29 ± 0.03 ) in PANC-1 cells with stable silencing FAM83A were significantly lower than those in the scrambled control group, respectively (1.95 ± 0.19, 0.98 ± 0.09;t=9.410, 12.600, P<0.05). After silencing FAM83A, the expression of stem cell marker CD133 (8.97 ± 0.62) and the sphere formation ability (8 ± 1) also decreased significantly compared with the scrambled group, respectively (21.60 ± 2.60, 25 ± 3; t=8.184, 9.311, P<0.05), and the stem cell-like phenotype of PANC-1 cells was also significantly inhibited. When PANC-1 cells were silenced by FAM83A and further treated with 50 μmol/L gecitabine at 72 h, the activity of FAM83A-silenced PANC-1 cells (32.33 ± 3.05)％ was significantly lower than that of the gecitabine alone treated group (63.06 ± 5.98)％ (t=6.378, P<0.05), and the apoptosis rate of FAM83A-silenced PANC-1 cells (76.52 ± 8.34) ％ was significantly higher than that of gemcitabine alone group (40.88 ± 4.91)％(t=7.929, P<0.05). After silencing FAM83A combined with IR irradiation, the activity of PANC-1 cells (43.25 ±4.21)％ was significantly lower than that of IR alone (78.13 ± 7.98)％ (t=6.694, P<0.05), and the apoptosis rate (44.56 ± 5.32)％ was significantly increased compared with IR alone (15.15 ±1.95)％ (t = 8.990, P < 0.05). After silencing FAM83A, the expression of Active-β-catenin was significantly decreased while the expression of p-β-catenin was significantly increased, the expression of β-catenin in the nucleus was significantly reduced, although total β-catenin had no significant change, and the activity of Wnt/β-catenin signaling pathway was significantly inhibited. Conclusions Silencing FAM83A could significantly reduce the stem cell-like traits and enhance the chemosensitivity and radiosensitivity of pancreatic cancer cells to gemcitabine and radiation via Wnt/β-catenin signaling pathway, which may provide a new target for targeted and combination therapy of pancreatic cancer.

Evaluation of the Cephalosporin Antimicrobial Drug Administration Program Using Monte Carlo Simulation / 中国药学杂志

OBJECTIVE: To evaluate and optimize the treatment regimen of cephalosporins based on Monte Carlo simulation and the pharmacokinetics /pharmacodynamics (PK/PD) model. METHODS: The treatment regimens of cefradine, cefuroxime, ceftriaxone and cefepime are 0.75 g qid, 1 g qid, 2 g bid and 2 g tid. And a total of 903 strains from eight species were collected. The minimum inhibitory concentration (MIC) was measured by trace broth dilution method. Monte Carlo simulation was used to simulate the regimens against Escherichia coli, Bacillus, Klebsiella pneumoniae, Acinetobacter baumannii, Citrobacter, Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa. Then, the cumulative fraction of response (CFR) was calculated. RESULTS: Cefhradine and cefuroximes' rates of resistance were higher (47.31% - 100%), the cumulative reaction scores(CFR) were all less than 90%. Cefuroxime has desirable regimen for Escherichia coli, Klebsiella pneumoniae, Citrobacter and Streptococcus pneumoniae. Cefepime has at least one dose regimen's CFR for more than 90%, and cephalosporins is consistent with time-dependent drug characteristics. CONCLUSION: For the treatment of infectious diseases, the choice of cephalosporins should be used to follow the principles of antibiotics. Target therapy should be used to test the pathogen susceptibility test, according to the value of MIC to simulate the corresponding dosing regimen, to achieve individualized administration.

Inhibition of FOXD1 gene expression increases radiosensitivity of colorectal cancer HCT116 cells / 中华放射医学与防护杂志

Objective To study the effect of inhibiting the expression of FOXD1 gene on the radiosensitivity of colorectal cancer cells. Methods The expressions of FOXD1 mRNA and protein in human colorectal cancer tissues and cells were detected by Real-time PCR ( qRT-PCR) and Western blot. The colorectal cancer cell line HCT116 was irradiated with 0, 2, 4 and 6 Gy of X-rays. The expression of FOXD1 in each groups were detected by qRT-PCR and Western blot. HCT116 cells were transfected with FOXD1 siRNA and its negative control and termed as si-FOXD1 group and si-NC group. When these cells were irradiated with 4 Gy X-rays, they were termed as si-FOXD1+4 Gy group and si-NC+4 Gy group. Cell proliferation was detected with MTT method, cell survival fraction was measured with colony formation assay, and DNA-PK activity was detected by TECT DNA-PK kit. The siRNA-transfected colorectal cancer cells were inoculated into BALB/c nude mice to establish the xenograft model. After irradiation, the volume and quality of the subcutaneous transplanted tumors were measured every 5 days. Results The expression of FOXD1 mRNA and protein in colorectal cancer tissues was higher than that in adjacent normal tissues (t=5. 579, 4. 816, P<0. 05). The mRNA(t=5. 85-17. 62, P<0. 05)and protein(t=9. 04-11. 42, P<0. 05) expression of FOXD1 in different colorectal cancer cell lines was higher than that in colonic mucosa epithelial cell line NCM460. The expression of FOXD1 in colorectal cancer cells HCT116 was increased after radiation in a dose dependent manner(t=9. 13-44. 15, P<0. 05). Transfection of si-FOXD1 effectively inhibited the expression of FOXD1 (t=10. 51, P<0. 05), decreased proliferation (t=10. 41, P <0. 05), increased radiosensitivity with a radiosensitization ratio of 1. 797, and reduced the radiation-induced DNA-PK activity ( t = 6. 20, P < 0. 05 ) in colorectal cancer cells. After localized irradiation, the tumor volume and weight in nude mice transplanted with si-FOXD1 HCT116 cells were significantly smaller than those in HCT116 (t=11. 29, 3. 69, P<0. 05). Conclusions Knock-down of FOXD1 gene increases the radiosensitivity of colorectal cancer cells and inhibits the growth of colorectal cancer xenograft in nude mice, which provides a potential target gene in improving the effect of radiotherapy on colorectal cancer.

Liver Tissue-related Metabolic Mechanism of Different Infusion Volumes for Hemorrhagic Shock / 法医学杂志

Objective To investigate the curative effects of various infusion volumes on liver-related metabolic mechanism in the treatment of hemorrhagic shock.Methods A severe hemorrhagic shock rabbit model was established in 30 rabbits.The rabbits were randomly divided into three groups:non-infusion group (A), conventional infusion group (B), and excessive infusion group (C) (n=10 in each group).Taking group B as the control, groups A and C were observed for the damage of non-infusion and excessive infusion, respectively.The outcomes in the three groups and their relations with liver tissue metabolism changes were analyzed with gas chromatograph-mass spectrometer (GC-MS).Results The mortality in groups A, B, and C group were 80％, 0％, and 70％, respectively.The liver tissue metabolic profile in group B showed statistically significant difference compared with that in groups A and B.In group C, the levels of 21 metabolites were lower than those in group B, and the levels of8 metabolites were lower than those in group A.The relative contents of various metabolites were correlated with infusion volumes, and the succinic acid content was associated with death events (P＜0.05).Conclusion The conventional infusion has significant curative effect on hemorrhagic shock.The metabolites of liver tissues with excessive infusion are generally decompensated and have longer survival time than those in non-infusion group, which may caused by the excessive infusion-induced blood volume increase after hemorrhagic shock.Tissue fluid dilution is an important cause of death.

Progress in research on the novel cancer therapeutic target Cdc20 / 药学学报

The anaphase promoting complex (APC) regulates cell cycle progression by forming two functionally distinct E3 ubiquitin ligase complexes, APCCdc20 activated by cell division cycle protein 20 (Cdc20) and APCCdh1 activated by Cdc20 homologue 1 (Cdh1), respectively. Cdc20 and Cdh1 have different functions in the occurrence and development of the tumor. Cdc20 is a cancer promoter while Cdh1 suppresses tumorigenesis. Emerging evidence has begun to reveal that Cdc20 has positive functions in tumorigenesis, the overexpression of Cdc20 has been observed in many cancers. Currently, Cdc20 inhibitors, mostly non-specific inhibitors except apcin, not only block the combination between Cdc20 and APC, also block the combination between Cdh1 and APC, which leads to a poor selectivity. In this paper, the Cdc20 role in the development and process of cancers and its inhibitors are reviewed.

Programmed cell death 4 enhances radiosensitivity of pancreatic cancer cells / 中华放射医学与防护杂志

Objective To investigate the effect and mechanism of programmed cell death 4 ( PDCD4 ) on radiosensitivity of pancreatic cancer cells. Methods Pancreatic cancer tissues and corresponding adjacent tissues were collected, the expression level of PDCD4 was detected by RT-PCR and Western blot. Human pancreatic cancer cells Sw1990 were transfected with PDCD4 overexpression vector ( group pIRES2-PDCD4 ) , empty vector ( pIRES2 group ) , and treated with transfection reagent, respectively. The expression level of PDCD4 was detected by RT-PCR and Western blot. After radiation treatment, cell apoptosis was detected by flow cytometry, cell survival was detected by clone assay, and the expression levels ofβ-catenin, c-myc and Cleaved Caspase-3 were detected by Western blot. Results The expression of PDCD4 mRNA and protein in pancreatic cancer tissues was significantly lower than that in adjacent tissues (t=4. 869, 9. 208, P<0. 05). The expression of PDCD4 mRNA and protein in pIRES2-PDCD4 group was significantly lower than that in the non-transfection group ( t =9. 074, 18. 927, P <0. 05). After radiation, the apoptosis rate and Cleaved Caspase-3 level in the pIRES2-PDCD4 group were significantly higher than those in the non-transfection group (t =3. 670, 4. 086, P <0. 05), while the expression levels of β-catenin and c-myc in the cells were significantly lower than those in the non-transfection group (t =9. 242, 17. 644, P <0. 05). The radiosensitivity of pIRES2-PDCD4 group was higher than that of non-transfection group, and the sensitization ratio was 1. 843. Conclusions PDCD4 can increase radiosensitivity and promote apoptosis of pancreatic cancer cells, to which the Wnt signaling pathway may be related.

Clinical Observation of Small Dose of Octreotide for Preventing Hyperamylasemia and Acute Pancreatitis after Endoscopic Retrograde Cholangiopancreatography / 中国药房

OBJECTIVE:To observe clinical effects and safety of small dose of octreotide for preventing hyperamylasemia and acute pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP).METHODS:One hundred and twenty ERCP pa tients were selected from our hospital during Oct.2014-Jan.2015 and then divided in to observation group and control group in accordance with random number table,with 60 cases in each group.Both groups were given Diazepam tablet 10 mg+Meperidine hydrochloride tablet 100 mg+Phenobarbital scopolamine tablet 2 tablets 0.5 h before surgery for sedation and analgesia,and routine acid suppression and anti-infective therapy.Observation group was additionally given Octreotide acetate injection 0.1 mg hypodermically and then given Octreotide acetate injection 0.1 mg immediately after surgery,8 h after surgery.The levels of serum amylase and blood glucose were observed in 2 groups,and the occurrence of postoperative complication and ADR were recorded.RESULTS:Before operation,there was no statistical significance in the levels of serum amylase and blood glucose between 2 groups (P＞ 0.05).After operation,the level of serum amylase in control group was significantly higher than in observation group,with statistical significance (P＜0.05).There was no statistical significance in blood glucose level between 2 groups after operation (P＞0.05).The incidence of hyperamylasemia and ADR in observation group was significantly lower than in control group,with statistical significance (P＜0.05),and there was no statistical significance in the incidence of acute pancreatitis between 2 groups after operation (P＞0.05).CONCLUSIONS:Small dose of octreotide can effectively reduce the level of serum amylase and the incidence of hyperamylasemia after ERCP with good safety.