Effects of non-muscle myosin Ⅱ silenced bone marrow-derived mesenchymal stem cells transplantation on lung extracellular matrix in rats after endotoxin/lipopolysaccharide-induced acute lung injury / 中华烧伤杂志

Objective: To investigate the effects of non-muscle myosin Ⅱ (NMⅡ) gene silenced bone marrow-derived mesenchymal stem cells (BMMSCs) on pulmonary extracellular matrix (ECM) and fibrosis in rats with acute lung injury (ALI) induced by endotoxin/lipopolysaccharide (LPS). Methods: The experimental research methods were adopted. Cells from femur and tibial bone marrow cavity of four one-week-old male Sprague-Dawley rats were identified as BMMSCs by flow cytometry, and the third passage of BMMSCs were used in the following experiments. The cells were divided into NMⅡ silenced group transfected with pHBLV-U6-ZsGreen-Puro plasmid containing small interference RNA sequence of NMⅡ gene, vector group transfected with empty plasmid, and blank control group without any treatment, and the protein expression of NMⅡ at 72 h after intervention was detected by Western blotting (n=3). The morphology of cells was observed by an inverted phase contrast microscope and cells labeled with chloromethylbenzoine (CM-DiⅠ) in vitro were observed by an inverted fluorescence microscope. Twenty 4-week-old male Sprague-Dawley rats were divided into blank control group, ALI alone group, ALI+BMMSC group, and ALI+NMⅡ silenced BMMSC group according to the random number table, with 5 rats in each group. Rats in blank control group were not treated, and rats in the other 3 groups were given LPS to induce ALI. Immediately after modeling, rats in ALI alone group were injected with 1 mL normal saline via tail vein, rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group were injected with 1×107/mL BMMSCs and NMⅡ gene silenced BMMSCs of 1 mL labelled with CM-DiⅠ via tail vein, and rats in blank control group were injected with 1 mL normal saline via tail vein at the same time point, respectively. At 24 h after intervention, the lung tissue was collected to observe intrapulmonary homing of the BMMSCs by an inverted fluorescence microscope. Lung tissue was collected at 24 h, in 1 week, and in 2 weeks after intervention to observe pulmonary inflammation by hematoxylin eosin staining and to observe pulmonary fibrosis by Masson staining, and the pulmonary fibrosis in 2 weeks after intervention was scored by modified Ashcroft score (n=5). The content of α-smooth muscle actin (α-SMA), matrix metalloproteinase 2 (MMP-2), and MMP-9 was detected by immunohistochemistry in 2 weeks after intervention (n=3), the activity of superoxide dismutase (SOD), malondialdehyde, myeloperoxidase (MPO) was detected by enzyme-linked immunosorbent assay at 24 h after intervention (n=3), and the protein expressions of CD11b and epidermal growth factor like module containing mucin like hormone receptor 1 (EMR1) in 1 week after intervention were detected by immunofluorescence staining (n=3). Data were statistically analyzed with one-way analysis of variance, Bonferroni method, and Kruskal-Wallis H test. Results: At 72 h after intervention, the NMⅡprotein expression of cells in NMⅡ silenced group was significantly lower than those in blank control group and vector group (with P values <0.01). BMMSCs were in long spindle shape and grew in cluster shaped like vortexes, which were labelled with CM-DiⅠ successfully in vitro. At 24 h after intervention, cell homing in lung of rats in ALI+NMⅡ silenced BMMSC group was more pronounced than that in ALI+BMMSC group, while no CM-DiⅠ-labelled BMMSCs were observed in lung of rats in blank control group and ALI alone group. There was no obvious inflammatory cell infiltration in lung tissue of rats in blank control group at all time points, while inflammatory cell infiltration in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group was significantly less than that in ALI alone group at 24 h after intervention, and alveolar wall turned to be thinner and a small amount of congestion in local lung tissue appeared in rats of the two groups in 1 week and 2 weeks after intervention. In 1 week and 2 weeks after intervention, collagen fiber deposition in lung tissue of rats in ALI alone group, ALI+BMMSC group, and ALI+NMⅡ silenced BMMSC group was significantly aggravated compared with that in blank control group, while collagen fiber deposition in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group was significantly improved compared with that in ALI alone group. In 2 weeks after intervention, modified Ashcroft scores for pulmonary fibrosis of rats in ALI alone group, ALI+BMMSC group, and ALI+NMⅡ silenced BMMSC group were 2.36±0.22, 1.62±0.16, 1.06±0.26, respectively, significantly higher than 0.30±0.21 in blank control group (P<0.01). Modified Ashcroft scores for pulmonary fibrosis of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group were significantly lower than that in ALI alone group (P<0.01), and modified Ashcroft score for pulmonary fibrosis of rats in ALI+NMⅡ silenced BMMSC group was significantly lower than that in ALI+BMMSC group (P<0.01). In 2 weeks after intervention, the content of α-SMA in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group were significantly decreased compared with that in ALI alone group (P<0.05 or P<0.01). The content of MMP-2 in lung tissue of rats in the 4 groups was similar (P>0.05). The content of MMP-9 in lung tissue of rats in ALI alone group was significantly increased compared with that in blank control group (P<0.01), and the content of MMP-9 in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group was significantly decreased compared with that in ALI alone group (P<0.01). At 24 h after intervention, the activity of malondialdehyde, SOD, and MPO in lung tissue of rats in ALI alone group, ALI+BMMSC group, and ALI+NMⅡ silenced BMMSC group were significantly increased compared with that in blank control group (P<0.01), the activity of malondialdehyde in lung tissue of rats in ALI+NMⅡ silenced BMMSC group and the activity of SOD in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group were significantly increased compared with that in ALI alone group (P<0.05 or P<0.01), and the activity of SOD in lung tissue of rats in ALI+NMⅡ silenced BMMSC group was significantly decreased compared with that in ALI+BMMSC group (P<0.01). The activity of MPO in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group was significantly decreased compared with that in ALI alone group (P<0.01), and the activity of MPO in lung tissue of rats in ALI+NMⅡ silenced BMMSC group was significantly decreased compared with that in ALI+BMMSC group (P<0.01). In 1 week after intervention, the protein expression of CD11b in lung tissue of rats in ALI+NMⅡ silenced BMMSC group was significantly increased compared with those in the other three groups (P<0.05 or P<0.01), while the protein expressions of EMR1 in lung tissue of rats in the four groups were similar (P>0.05). Conclusions: Transplantation of NMⅡ gene silenced BMMSCs can significantly improve the activity of ECM components in the lung tissue in LPS-induced ALI rats, remodel its integrity, and enhance its antioxidant capacity, and alleviate lung injury and pulmonary fibrosis.

The present situation and progress of treatment of acute type A aortic dissection with organ malperfusion syndrome / 中华胸心血管外科杂志

With the development of brain protection, management of aortic root and arch lesions, the surgical mortality of acute type A aortic dissection is gradually decreasing, and malperfusion syndrome(MPS) is becoming increasingly prominent as A major problem affecting the short-term efficacy of acute type A aortic dissection. Aortic dissection may lead to poor perfusion of various organs, among which brain, heart, abdominal organs, kidney, lower limbs and spinal cord are the most prominent, with a total incidence of about 30%. There was a significant difference in surgical mortality between aortic dissection with or without organ MPS. As the number of organs involved increased, the mortality rate increased significantly. Abdominal viscera, especially mesenteric malperfusion is the key point of the surgical attention. At present, the contents worth discussing are as follows : (1) the definition of organ malperfusion syndrome of aortic dissection; (2) accurate classification or classification of poor perfusion of each system; (3)establish a more accurate scoring system for preoperative risk assessment of aortic dissection; (4)Thoracic Endovascular Aortic Repair priority or time priority or individualization should be adopted for the management of organ perfusion syndrome.

IL-7 regulates in vitro activity of CD8 + T cells in patients with sepsis through modulation of CD127 expression / 中华微生物学和免疫学杂志

Objective:To investigate the expression of membrane-bound CD127 (mCD127) and soluble CD127 (sCD127) in patients with sepsis, and to assess the mechanism of IL-7 in regulating CD8 + T cell activity in these patients. Methods:A prospective cohort study was conducted on 47 patients with sepsis (sepsis group) and 18 healthy controls (control group). Serum samples and peripheral blood mononuclear cells (PBMCs) were isolated. CD8 + T cells were purified. IL-7 and sCD127 levels in serum were measured by enzyme-linked immunosorbent assay. Expression of mCD127 on CD8 + T cells was measured by flow cytometry. Total CD127 and sCD127 expression at mRNA level in CD8 + T cells was semi-quantified by real-time PCR. CD8 + T cells were stimulated with recombinant human IL-7, along with signal transducer and activator of transcription 5 (STAT5) inhibitor or phosphatidylinositol 3-kinase (PI3K) inhibitor. Changes in mCD127 expression on CD8 + T cells and the expression of total CD127 and sCD127 at mRNA level were then measured. The cytotoxicity of CD8 + T cells in response to IL-7 stimulation was assessed using a co-culture system with CD8 + T cells and MCF-7 cells. Student′s t test and LSD- t test were used for statistical analysis. Results:Serum IL-7 and sCD127 were lower in sepsis group than in control group [(101.82±12.58) pg/ml vs (111.07±11.10) pg/ml, P<0.01; (278.58±62.31) pg/ml vs (334.62±70.55) pg/ml, P<0.01]. Serum IL-7 was positively correlated with serum sCD127 in patients with sepsis ( r=0.46, P<0.01). The percentage of mCD127 + CD8 + T cells in CD8 + T cells and the mean fluorescence intensity of mCD127 in sepsis group were higher than those in control group ( P<0.05). The expression of sCD127 at mRNA level in CD8 + T cells was lower in sepsis group than in control group (1.34±0.33 vs 1.80±0.60, P<0.001). Stimulation with recombinant human IL-7 promoted sCD127 secretion and total CD127 and sCD127 expression at mRNA level in CD8 + T cells ( P<0.05). Inhibition of STAT5 suppressed the IL-7-induced sCD127 secretion and total CD127 and sCD127 expression at mRNA level ( P<0.05). However, inhibition of PI3K could not achieve those effects ( P>0.05). CD8 + T cells-induced target cell death was inhibited in sepsis group as compared with that in control group [(12.49±2.12)% vs (23.83±3.76)%, P<0.001]. Recombinant human IL-7 promoted the CD8 + T cell-induced target cell death ( P<0.05) and increased the secretion of cytokines and cytotoxic granule proteins ( P<0.05). Inhibition of STAT5 suppressed IL-7-mediated CD8 + T cell cytotoxicity ( P<0.05). However, inhibition of PI3K did not affect IL-7-mediated CD8 + T cell cytotoxicity ( P>0.05). Conclusions:IL-7 promoted sCD127 secretion and enhanced the in vitro cytotoxicity of CD8 + T cells in patients with sepsis through STAT5 signal pathway.

Research progress of Nrf2 inhibitor in tumor therapy / 药学学报

Nrf2 is a multi-effect transcription factor, which plays a crucial role in cytoprotective system. With the deepening of research on new regulatory modes and biologic functions of Nrf2, the oncogenic role of Nrf2 in malignant transformed tumors is increasingly obvious. More and more evidences show that Nrf2 is involved in the whole process of tumor occurrence, development, metastasis and prognosis, and inhibiting Nrf2 may be a promising strategy in tumor therapy. However, the development of Nrf2 inhibitors is still in early stage. In this paper, the biological function of Nrf2 and its dual role in tumor are briefly introduced, and representative Nrf2 inhibitors are reviewed according to their structure types, so as to provide reference and ideas for the development of anti-tumor drugs centering on the regulation of Nrf2.

Overexpression of NAT10 induced platinum drugs resistance in breast cancer cell / 中华肿瘤杂志

Objective: To observe the platinum drugs resistance effect of N-acetyltransferase 10 (NAT10) overexpression in breast cancer cell line and elucidate the underlining mechanisms. Methods: The experiment was divided into wild-type (MCF-7 wild-type cells without any treatment) group, NAT10 overexpression group (H-NAT10 plasmid transfected into MCF-7 cells) and NAT10 knockdown group (SH-NAT10 plasmid transfected into MCF-7 cells). The invasion was detected by Transwell array, the interaction between NAT10 and PARP1 was detected by co-immunoprecipitation. The impact of NAT10 overexpression or knockdown on the acetylation level of PARP1 and its half-life was also determined. Immunostaining and IP array were used to detect the recruitment of DNA damage repair protein by acetylated PARP1. Flow cytometry was used to detect the cell apoptosis. Results: Transwell invasion assay showed that the number of cell invasion was 483.00±46.90 in the NAT10 overexpression group, 469.00±40.50 in the NAT10 knockdown group, and 445.00±35.50 in the MCF-7 wild-type cells, and the differences were not statistically significant (P>0.05). In the presence of 10 μmol/L oxaliplatin, the number of cell invasion was 502.00±45.60 in the NAT10 overexpression group and 105.00±20.50 in the NAT10 knockdown group, both statistically significant (P<0.05) compared with 219.00±31.50 in wild-type cells. In the presence of 10 μmol/L oxaliplatin, NAT10 overexpression enhanced the binding of PARP1 to NAT10 compared with wild-type cells, whereas the use of the NAT10 inhibitor Remodelin inhibited the mutual binding of the two. Overexpression of NAT10 induced PARP1 acetylation followed by increased PARP1 binding to XRCC1, and knockdown of NAT10 expression reduced PARP1 binding to XRCC1. Overexpression of NAT10 enhanced PARP1 binding to LIG3, while knockdown of NAT10 expression decreased PARP1 binding to LIG3. In 10 μmol/L oxaliplatin-treated cells, the γH2AX expression level was 0.38±0.02 in NAT10 overexpressing cells and 1.36±0.15 in NAT10 knockdown cells, both statistically significant (P<0.05) compared with 1.00±0.00 in wild-type cells. In 10 μmol/L oxaliplatin treated cells, the apoptosis rate was (6.54±0.68)% in the NAT10 overexpression group and (12.98±2.54)% in the NAT10 knockdown group, both of which were statistically significant (P<0.05) compared with (9.67±0.37)% in wild-type cells. Conclusion: NAT10 overexpression enhances the binding of NAT10 to PARP1 and promotes the acetylation of PARP1, which in turn prolongs the half-life of PARP1, thus enhancing PARP1 recruitment of DNA damage repair related proteins to the damage sites, promoting DNA damage repair and ultimately the survival of breast cancer cells.

Dynamic control of ERG20 expression to improve production of monoterpenes by engineering Saccharomyces cerevisiae / 中国中药杂志

Monoterpenes are widely used in cosmetics, food, medicine, agriculture and other fields. With the development of synthetic biology, it is considered as a potential way to create microbial cell factories to produce monoterpenes. Engineering Saccharomyces cerevisiae to produce monoterpenes has been a research hotspot in synthetic biology. In S. cerevisiae, the production of geranyl pyrophosphate(GPP) and farnesyl pyrophosphate(FPP) is catalyzed by a bifunctional enzyme farnesyl pyrophosphate synthetase(encoded by ERG20 gene) which is inclined to synthesize FPP essential for yeast growth. Therefore, reasonable control of FPP synthesis is the basis for efficient monoterpene synthesis in yeast cell factories. In order to achieve dynamic control from GPP to FPP biosynthesis in S. cerevisiae, we obtained a novel chassis strain HP001-pERG1-ERG20 by replacing the ERG20 promoter of the chassis strain HP001 with the promoter of cyclosqualene cyclase(ERG1) gene. Further, we reconstructed the metabolic pathway by using GPP and neryl diphosphate(NPP), cis-GPP as substrates in HP001-pERG1-ERG20. The yield of GPP-derived linalool increased by 42.5% to 7.6 mg·L~(-1), and that of NPP-derived nerol increased by 1 436.4% to 8.3 mg·L~(-1). This study provides a basis for the production of monoterpenes by microbial fermentation.

Construction of cell factories for high production of ginsenoside Rh_2 in Saccharomyces cerevisiae / 中国中药杂志

Ginsenoside Rh_2 is a rare active ingredient in precious Chinese medicinal materials such as Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Panacis Quinquefolii Radix. It has important pharmacological activities such as anti-cancer and improving human immunity. However, due to the extremely low content of ginsenoside Rh_2 in the source plants, the traditional way of obtaining it has limitations. This study intended to apply synthetic biological technology to develop a cell factory of Saccharomyces cerevisiae to produce Rh_2 by low-cost fermentation. First, we used the high protopanaxadiol(PPD)-yielding strain LPTA as the chassis strain, and inserted the Panax notoginseng enzyme gene Pn1-31, together with yeast UDP-glucose supply module genes[phosphoglucose mutase 1(PGM1), α-phosphoglucose mutase(PGM2), and uridine diphosphate glucose pyrophosphorylase(UGP1)], into the EGH1 locus of yeast chromosome. The engineered strain LPTA-RH2 produced 17.10 mg·g~(-1) ginsenoside Rh_2. This strain had low yield of Rh_2 while accumulated much precursor PPD, which severely restricted the application of this strain. In order to further improve the production of ginsenoside Rh_2, we strengthened the UDP glucose supply module and ginsenoside Rh_2 synthesis module by engineered strain LPTA-RH2-T. The shaking flask yield of ginsenoside Rh_2 was increased to 36.26 mg·g~(-1), which accounted for 3.63% of the dry weight of yeast cells. Compared with those of the original strain LPTA-RH2, the final production and the conversion efficiency of Rh_2 increased by 112.11% and 65.14%, respectively. This study provides an important basis for further obtaining the industrial-grade cell factory for the production of ginsenoside Rh_2.

Short needling for knee osteoarthritis with blood stasis obstruction and its effect on serum inflammatory factors / 中国针灸

OBJECTIVE@#To compare the clinical efficacy and its effect on serum levels of tumor necrosis factor α(TNF-α), interleukin 1β(IL-1β), interleukin 6 (IL-6) and prostaglandin E2 (PGE2) between short needling (close-to-bone needling) and conventional acupuncture for knee osteoarthritis (KOA) with blood stasis obstruction.@*METHODS@#A total of 68 KOA patients with blood stasis obstruction were randomized into a short needling group (34 cases, 3 cases dropped off) and a conventional acupuncture group (34 cases, 3 cases dropped off). The same acupoints (Dubi [ST 35], Neixiyan [EX-LE 4], Binzhong [Extra], Liangqiu [ST 34], etc. on the affected side) were selected in the two groups. In the short needling group, short needling technique was adopted, the needles were slowly inserted and the needle bodies were shaken, thus gradually penetrated to the bone. In the conventional acupuncture group, conventional acupuncture was adopted, the needles were penetrated to the muscle. After qi-arrival, Dubi (ST 35) and Neixiyan (EX-LE 4), Zusanli (ST 36) and Liangqiu (ST 34) were connected with CMNS6-1 electronic acupuncture instrument, with disperse-dense wave, 2 Hz/10 Hz in frequency, the current intensity was based on patients' feeling, the needles were retained for 30 min, at the same time, the knee joint was irradiated for 30 min with a special electromagnetic wave apparatus in the two groups. Once every other day, 3 times a week for 4 weeks. Before and after treatment, the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) score, knee joint pain visual analogue scale (VAS) score, inflammatory response related indexes (serum TNF-a, IL-1β, IL-6 and PGE2) and knee joint ultrasound were observed，and the clinical effect was evaluated in the two groups.@*RESULTS@#After treatment，the pain, stiffness, function scores and total scores of WOMAC were decreased as compared with those before treatment in the two groups (P<0.05), except for the pain score, the changes of above scores in the short needling group were greater than the conventional acupuncture group (P<0.05). After treatment, the VAS scores, serum levels of TNF-a, IL-1β, IL-6, PGE2 and knee joint synovium thickness, intra-articular effusion were decreased as compared with those before treatment in the two groups (P<0.05), the levels of TNF-a, IL-1β, IL-6 in the short needling group were lower than the conventional acupuncture group (P<0.05). The total effective rate in the short needling group was 87.1% (27/31), which was superior to 83.9% (26/31) in the conventional acupuncture group (P<0.05).@*CONCLUSION@#Short needling could improve the knee joint function, relieve the pain and inflammatory response, improve the knee joint synovium inflammatory response, reduce the knee joint intra-articular effusion for KOA patients, its effect is better than conventional acupuncture.

Whole-genome sequencing of SARS-CoV-2 Gamma variant first discovered in Chinese mainland / 中华微生物学和免疫学杂志

Objective:To investigate the whole genome of SARS-CoV-2 causing COVID-19 in Rongcheng city of Shandong Province in May 2022 and to further analyze the nucleotide and amino acid variations for source tracing.Methods:High-throughput sequencing was used to sequence the SARS-CoV-2 genome in 15 nasopharyngeal swab samples from COVID-19 cluster infections and three environmental samples related to an aquatic product import company. Whole-genome sequence splicing, variant site analysis and sequence typing were performed on the raw sequencing data using virus sequence and variant analysis software. A phylogenetic tree was constructed by evolutionary analysis software. Epidemiological investigation was used to trace the potential source of infection.Results:Thirteen whole genome sequences of SARS-CoV-2 with the length ranging from 29 653 bp to 29 780 bp were successfully obtained from the nasopharyngeal swab samples. The average sequencing depth was 1 756-6 565 X and the genome coverage was 99.20%-99.63%. The results of Pangolin typing showed that the 13 genomes belonged to the VOC/Gamma (P.1.15) evolutionary branch. Compared with the Wuhan reference strain (NC_045512.2), the 13 genome sequences had 40-41 nucleotide mutation sites. There were 23-24 amino acid variation sites in seven protein domains (ORF1a, ORF1b, S, ORF3a, ORF8, ORF9b and N proteins). Evolutionary analysis showed that the viral sequence was grouped to the same subclade as the reference strain from Argentina (EPI_ISL_4082233).Conclusions:In this study, the whole genome sequences of 13 Gamma variant strains were obtained from COVID-19 cluster infections associated with imported cold-chain aquatic products in Rongcheng city, and the imported seafood from South America in 2021 was found to be the source of the virus in a timely manner. This study provided reference for the SARS-CoV-2 variant analysis and case tracing and also suggested that the survival and transmission ability of SARS-CoV-2 on the surface of cold-chain products should not be underestimated and needed further investigation.

Exploratory research of the value of abbreviated comprehensive geriatric assessment in elderly breast cancer patients / 中华老年医学杂志

Objective:To explore the value and feasibility of abbreviated comprehensive geriatric assessment(aCGA)grading in elderly patients with breast cancer.Methods:From June 2019 to January 2020, elderly patients with breast cancer aged 65 years and above were enrolled.Eastern Cooperative Oncology Group(ECOG)score and aCGA classification were performed respectively.Clinical characteristics, score distribution and differences between the two assessment methods were compared and analyzed.Results:A total of 61 cases of breast cancer patients aged 65 years and above were included in our study.According to the assessment of aCGA, grade A accounted for 65.5%(40/61), grade B accounted for 27.9%(17/61), and grade C accounted for 6.6%(4/61), among which 82.0%(50/61 cases)of the patients had complications.And the most common complications were hypertension, cardiovascular disease and diabetes.Among the 50 patients with ECOG score 0-1, 74.0%(37/50)were aCGA grade A, and 26.0%(13/50)were aCGA grade B.Conclusions:According to the aCGA grading, about two thirds of breast cancer patients over 65 years old are assessed as grade A, which indicates that they might have better tolerance during the treatment.However, among the patients with 0-1 score according to the ECOG score, some patients still have a slightly worse grade(aCGA grade B, which shows slightly worse health condition), suggesting that the refinement degree of ECOG score may be insufficient, and the health damage of some patients may be underestimated.

Phenylpropanoids from Zanthoxylum species and their pharmacological activities: a review / 中国中药杂志

Phenylpropanoids are one of the major chemical constituents in Zanthoxylum species. They include simple phenylpropanoids, coumarins, and lignans and possess anti-tumor, anti-inflammatory, anti-platelet aggregation, anti-bacterial, anti-viral, insecticidal, and antifeedant activities. This review summarizes the chemical constituents and pharmacological activities from the Zanthoxylum plants in hopes of providing reference for the research and application of phenylpropanoids from this genus.

Clinical genetic analysis and diagnosis of a family with hereditary hemorrhagic telangiectasia / 中华耳鼻咽喉头颈外科杂志

Objective: To explore the diagnostic significance of the combination of clinical and genetic detection of hereditary hemorrhagic telangiectasia (HHT) by analyzing the clinical and genetic diagnosis of a family with HHT. Methods: Medical history data of the probands and their family members were collected, and the sequence analyses of coding regions of ENG, ACVRL1, SMAD4 and GDF2 genes were performed by PCR-sequencing method, and a comprehensive diagnosis was made based on the clinical features and gene detection results. After the pathogenic gene variation was identified, 11 members of 3 generations of the family were tested for pathogenic gene mutation. Results: There was an ACVRL1 c.715_716delAG mutation in the proband and 9 other family members, which caused p.S239C. Based on the clinical and genetic findings, the 7 suspected were diagnosed and 2 asymptomatic patients were found to carry the mutation site. Conclusion: The combination of clinical features and gene detection can determine the etiology and classification of HHT, which is convenient for the early diagnosis and prevention of the disease.

Effect of neuregulin-1 on cardiac glucose metabolism in rats with experimental myocardial infarction / 中华心血管病杂志

Objective: To investigate the effect of neuregulin-1(NRG-1) on cardiac glucose metabolism in Sprague Dawley (SD) rats with experimental myocardial infarction (MI). Methods: Adult male SD rats were randomly divided into three groups: the sham-operated group, MI group, and MI+NRG1 group. The rat MI model was established via ligation of the left anterior descending coronary artery. Two weeks after operation, echocardiography was performed, MI rats with left ventricular ejection fraction (LVEF) between 0.3-0.5 were selected and randomly assigned to MI group and MI+NRG-1 group. Rats in MI+NRG-1 group were treated with recombinant human NRG-1β (100 μg/kg) via tail vein at 2 weeks after operation (twice per week for 6 weeks); while rats in sham-operated group and MI group received equal volume of physiological saline. By the end of administration, echocardiography and small animal positron emission tomography (PET) were performed to detect cardiac function and myocardial glucose uptake. Myocardial morphology and collagen volume fraction, cardiomyocyte apoptosis and reactive oxygen species (ROS) production were evaluated by histopathologic analysis. Myocardial pyruvate dehydrogenase (PDH) and citrate synthase (CS) activity, as well as ATP production were detected by commercial kits. The mRNA and protein expression levels of NRG-1, p-ErbB4, and key factors involved in glucose metabolism (including Glut-4, HK2, PDK4, PDH, CS) were detected by quantitative real-time PCR (qRT-PCR) and Western blot assay, respectively. Results: With the MI model successfully established, the left ventricular ejection fraction(LVEF) and left ventricular shortening fraction(LVFS) were significantly lower in MI group and MI+NRG-1 group than that in sham group (both P<0.01), while there was no significant difference between MI group and MI+NRG-1 group(all P>0.05). After 6 weeks of NRG-1β intervention, the LVEF and LVFS were significantly higher in MI+NRG-1 group than in MI group (both P<0.01). By the end of experiment, PET imaging showed that the mean standardized uptake value (SUVmean) were lower in MI+NRG-1 group than in the sham group (4.06±0.28 vs. 5.18±0.37, P<0.01), while significantly higher than that in MI group (4.06±0.28 vs.2.86±0.49, P<0.01). Histopathological analysis showed that compared with MI group, rats in MI+NRG-1 group exhibited significantly decreased left ventricle collagen volume fraction ((7.83±1.24) % vs. (18.31±3.58) %, P<0.01), cardiomyocyte apoptosis((37.98±4.26)% vs. (67.04±5.38)%, P<0.01), and DHE fluorescence intensity(0.057 28±0.007 06 vs. 0.076 94±0.008 46, P<0.01), indicating that NRG-1β could reduce ROS production. PDH activity, CS activity, and ATP production were significantly higher in MI+NRG-1 group than in MI group (all P<0.05). qRT-PCR demonstrated an upregulated Glut-4, HK2 and CS, but downregulated PDK4 mRNA expression in MI+NRG-1 group compared with MI group (all P<0.01). Western blot assay showed significantly higher protein expression of NRG-1, p-ErbB4, Glut-4, HK2, PDH, CS in MI+NRG-1 group than in MI group (all P<0.01). Conclusion: NRG-1 could improve glucose uptake and utilization in myocardium by activating phosphorylation of myocardial ErbB4 receptor in MI rats, thus providing a therapeutic option for improving energy metabolism after MI.

Research Progress of Gypsum Ustum and Application of Calcium-containing Hemostatic Materials / 中国实验方剂学杂志

Mineral medicine is an indispensable part of traditional Chinese medicine and has a long history of application. Among them， mineral-based hemostatics have been widely applied for the treatment of various hemorrhagic diseases with extensive clinical experience and established efficacy. Gypsum Fibrosum （GF）， a commonly used mineral medicine in clinical， can clear away heat， and relieve anxiety and thirst. Gypsum Ustum （GU） is the processed product of GF after calcining at high temperature. It is mainly composed of anhydrous calcium sulfate （CaSO4） with the functions of moisturizing， promoting muscle growth， astringent sores and hemostasis. GU is often used externally to treat ulcer， itching， eczema， water and fire scalds， trauma bleeding， etc. Studies on the mechanism of hemostasis have shown that Ca2+ （coagulation factor Ⅳ） is involved in many key processes of the internal and external coagulation cascades and can prevent bleeding by regulating platelet activation and aggregation， and promoting the production of insoluble fibrin and the ultimate formation of a blood clot. GF and GU both contain Ca2+ which provide an important material basis of hemostatic effect for both compounds， but GU has a significant hemostatic effect， while GF has no hemostatic effect. After processing， the taste and efficacy of the GF have been obviously changed which reflects the characteristics of processing， but the processing mechanism of GU has not been fully clarified. Therefore， based on studies of GF before and after calcining， this paper focused on these aspects including calcining process， crystal form comparison， element content， efficacy comparison， and summarized various aspects of Ca2+ involved in hemostasis. In addition， the hemostatic properties of other calcium-containing mineral medicines and new calcium-containing hemostatic materials such as calcium alginate， mesoporous calcium silicate and nanogel hemostatic materials were also discussed. The paper aimed to provide a reference for elucidating processing mechanism and clinical dialectical use of GU， also to promote development of new calcium-containing hemostatic materials.

Repeatability of OPD-Scan Ⅲ and its consistency with Pentacam in measuring keratometry and astigmatism / 中华实验眼科杂志

Objective:To analyze the repeatability of keratometry and astigmatism values measured by the OPD-Scan Ⅲ and the agreement of the parameters measured by OPD-Scan Ⅲ and Pentacam.Methods:A diagnostic test study design was adopted.Fifty patients (100 eyes) with refractive errors, aged from 21 to 35 years old, were selected from Second Affiliated Hospital of Nantong University and Lixiang Eye Hospital of Soochow University during August 2018.Spherical equivalent, astigmatim degree and axis were measured by Autorefraction.Corneal biometric measurements were measured three times continuously with the above two instruments.Keratometry values at the flat axis (K1), keratometry values at the steep axis (K2), astigmatim degree, axis, vector parameters J0 (Jackson cross cylinder at 0°or 180°) and J45 (Jackson cross cylinder at 45°) were recorded.Intraclass correlation coefficient (ICC) was used for repeatability analysis.Wilcoxon signed rank test, Spearman correlation analysis and Bland-Altman graphs were employed to analyze the comparability.This study adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Lixiang Eye Hospital of Soochow University (No.SLER2018112). Written informed consent was obtained from each patient prior to any examination.Results:The ICC of K1, K2, astigmatism, astigmatic axis, J0 and J45 measured by OPD-Scan Ⅲ were all greater than 0.900; the ICC of the astigmatism measured by Pentacam was 0.896, and the ICC of the other parameters measured by Pentacam were greater than 0.900; The values of K2, astigmatism, J0 and J45 measured by OPD-Scan Ⅲ were greater than those measured by Pentacam, and the differences were statistically significant (all at P<0.05). The values of K1, K2, astigmatism degree, axis, J0 and J45 measured by OPD-Scan Ⅲ were positively correlated with those measured by Pentacam (r s=0.981, 0.982, 0.900, 0.737, 0.921, 0.703, all at P<0.01). The 95% agreement of limits (LOA) of K1, K2, astigmatism, axis, J0 and J45 measurement difference between OPD-Scan Ⅲ and Pentacam were -0.52-0.50 D, -0.39-0.59 D, -0.37-0.48 D, -17.29°-20.38°, -0.12-0.24 D and -0.22-0.28 D, respectively. Conclusions:OPD-Scan Ⅲ has high credibility in measuring corneal refractive power and astigmatism degree, but its 95% LOA of astigmatism axis is too large to be accepted clinically.

Advances in diagnosis and surgical treatment of infective endocarditis / 中国胸心血管外科临床杂志

@#Infective endocarditis (IE) is a disease with severe complications and high mortality. It is heterogeneous in etiology, clinical manifestations, and course. At the same time, there are many disputes on the clinical practice of antibiotic treatment, surgical indications and timing. In this review, we discuss the epidemiology, diagnosis, treatment, and prevention of IE, especially the latest advances in surgical treatment after the release of European Society of Cardiology and American Heart Association guidelines in 2015.

Effect of different orthodontic forces on the expression of T helper cell 17 cell-related cytokines in the pressure side of periodontal tissue in rats / 华西口腔医学杂志

OBJECTIVES@#This study aimed to explore the changes in the expression of the characteristic transcription factor retinoid related orphan receptor γt (RORγt) and the cytokine interleukin-17 (IL-17) of T helper cell 17 (Th17) in the pressure side of the periodontal tissue of rats under different orthodontic forces. Their effects on the expression of osteoprotegerin (OPG) and the quantity of osteoclast (OC) were also explored. The role of Th17 cell in alveolar bone remodeling under different forces was preliminarily investigated.@*METHODS@#A total of 108 rats were chosen and randomly divided into three groups. Mesial forces of 0, 50, and 100 g were loaded on the maxillary first molar in the three groups. The rats were executed at 0, 1, 3, 5, 7, and 14 days. The expression of RORγt mRNA was quantified by real-time quantitative polymerase chain reaction. The expression of IL-17 protein was quantified by enzyme linked immunosorbent assay. The expression levels of RORγt and OPG proteins were quantified, and the quantity of OC was counted via immunohistochemistry.@*RESULTS@#The expression levels of RORγt and IL-17 and the quantity of OC increased first and then decreased in the 50 and 100 g groups, and the peak values of the two groups were on days 5 and 7, respectively. The expression levels in the 50 g group basically recovered to normal level on day 14, while that in the 100 g group remained at a high level. The expression levels in the 50 g group were higher than those in the 0 g group and lower than those in the 100 g group. The expression of OPG in the 50 g group decreased first, then increased, and finally decreased. It basically recovered to normal level on day 14. The expression of OPG in the 100 g group decreased first and then increased. It remained at a high level on day 14. The expression in the 50 g group was significantly higher than that in the 0 g group on day 7, while the expression in the 100 g group was significantly higher than that in the 0 g group on day 14.@*CONCLUSIONS@#RORγt, IL-17, and OPG were expressed regularly over time under different orthodontic forces, indicating that Th17 participated in the process of bone resorption on the pressure side of periodontal tissue by secreting IL-17.

Optogenetic study of glutamatergic neuron in parasubiculum in visuospatial memory / 中华行为医学与脑科学杂志

Objective:To investigate the function of glutamatergic neuron of the parasubiculum in spatial memory.Methods:Sixteen adult male C57BL/6 mice aged 6-8 weeks were divided into two groups randomly, 0.4 μl AAV5-CaMKⅡα-eNpHR3.0-eYFP was injected into the bilateral parasubiculum respectively in experimental group, equal dose AAV5-CaMKⅡα-eYFP for control group.The optic fiber was implanted 6 weeks after virus injection.The novel object place recognition test was performed one week after optic fiber implantation, continuous yellow light was delivered during the behavioral test to inhibit the function of glutamatergic neuron in the parasubiculum.The standard memory index (D2) was used to evaluate the spatial memory function.SPSS 20.0 software was used to process the data, and the independent-samples t-test and paired-samples t-test were used for data analysis. Results:In the novel object place recognition experiment, the mice showed no preference for either object in both control group(new object: 0.51±0.06, familiar object: 0.49±0.04, t=1.21, P>0.05) and experimental group(new object: 0.49±0.05, familiar object: 0.50±0.04, t=-0.78, P>0.05). Compared with the control group (0.55±0.06), the D2 score of the experimental group (0.26±0.07) was significantly lower ( t=-2.96, P<0.05), and the number of c-fos positive neuron in experimental group (96.33±7.13) was also significantly less than that in control group (127.67±5.24, t=-3.54, P<0.05). Conclusion:Inhibiting glutamatergic neuronal activity in the parasubiculum impairs spatial memory in mice, suggesting that glutamatergic neurons of the parasubiculum play an important role in spatial memory.

Preparation and in vivo and in vitro Characteristic Evaluation of Ligustrazine Ophthalmic Liposome Thermosen- sitive Gel / 中国药房

OBJECTIVE：To prepare Liguatrazine opthalmic liposome therm osensitive gel ，and to investigate its in vivo and in vitro characteristics. METHODS ：The ammonium sulfate gradient method was used to prepare Liguatrazine liposomes. The preparation technology was optimized by using orthogonal test. Using poloxamer P 407 as gel matrix ，Liguatrazine liposomes were prepared into thermosensitive gel. A membraneless model was used to study the dissolution and in vitro drug release of the gel. The modified Franz diffusion cell was used to investigate corneal permeability and further determine corneal hydration value. The effects of the gel on the proliferation of human corneal epithelial cell HCE-T. HE staining and Draize test were used to investigate the stimulatory effects of the gel on corneal cells of the rabbit ，and the histological changes of the eyes were observed. RESULTS ：The optimal preparation technology of Liguatrazine liposome was drug-lipid ratio of 1 ∶ 10（m/m），the ammonium sulfate concentration of 0.2 mol/L，phospholipid-cholesterol ratio of 4∶1（m/m），incubation temperature of 45 ℃. Then ligustrazine opthalmic liposome thermosensitive gel was prepared with 23％ poloxamer P 407 as gel matrix. The gel had good gelatinization temperature. The in vitro drug release and dissolution showed zero-order kinetic characteristics ，and in vitro drug release of the gel was mainly related to dissolution （R2＝0.993 4）. The cumulative transcorneal permeability of the gel was 43.3％ within 6 hours and corneal hydration value was 72.98％. Low and medium concentrations （1，5 mg/L）of Ligustrazine opthalmic liposome thermosensitive gel had no obvious proliferation toxicity to HCE-T cells ，but it showed cytotoxicity at high concentration （10 mg/L）. The mean Draize eyeirritation score of the gel on rabbit cornea was within non-stimulation，and there was no abnormal change in rabbit （No.2018001） corneal histology. CONCLUSIONS ： Prepared Ligustrazine opthalmic liposome thermosensitive gel has a suitable phase transition temperature ，good corneal permeability ，and low corneal irrit ation.

Application Value of Abbreviated Comprehensive Geriatric Assessment in Elderly Female Breast Cancer Patients / 中国医学科学院学报

Objective To evaluate the application value of abbreviated comprehensive geriatric assessment(aCGA)in elderly female breast cancer patients. Methods Eight aspects of the traditional CGA were simplified to form the aCGA assessment table,based on which the patients were classified into three grades of A,B and C according to the total scores.This study enrolled the elderly female patients with breast cancer aged 70 years and above who were treated in PUMC Hospital from June 2018 to January 2020.Eastern Cooperative Oncology Group(ECOG)scoring and aCGA grading were performed respectively,and the results of the two methods were compared. Results Of the 162 patients,111(68.5%)were classified by the aGGA method as grade A,43(26.5%)as grade B,and 8(5.0%)as grade C;131(80.9%)cases have concurrent diseases,and the most common complications were hypertension(