Objective To study the chemical constituents of Zanthoxylum dimorphophyllum and identify the chemical structures.Methods The compounds were isolated by silica gel,Flash column chromatography and purified by crystallization.Their structures were elucidated by spectral methods.ResultsSeven compounds were isolated and identified.They are 6-(3′,methyl-2′,3′-dihydroxy) butyl-7-methoxyl-8-(3″-methyl-2″-butenyl)-coumarin(Ⅰ),8-(3′-methyl)-2′,3′-butenyl-2′-(1″-hydroxy-1″-methyl)-ethyl-6,7-dihydrofurancoumarin(Ⅱ),6,7,8-trimethoxycoumarin(Ⅲ),scoparone(Ⅳ),umbelliferone(Ⅴ),canthin-6-one(Ⅵ),and syringaresinol(Ⅶ).Conclusion Compound I is a novel compound and all compounds are obtained from Z.dimorphophyllum for the first time.

To study the chemical constituents of the fruits of Melia toosendan, three limonoids were isolated and purified by repeated silica gel column chromatography and preparative HPLC from the EtOAc extract of M. toosendan. Their structures were determined by their physico-chemical properties and spectroscopic data (1D-NMR, 2D-NMR) as: 24, 25, 26, 27-tetranorapotirucalla-(apoeupha)-1alpha-tigloyloxy-3alpha, 7alpha-dihydroxyl-12alpha-acetoxyl-14, 20, 22-trien-21, 23-epoxy-6, 28-epoxy (1), nimbolinin B (2), and trichilinin D (3), separately. Compound 1 is a new compound, and compound 2 is obtained from this plant for the first time.

Objective To investigate the effect and mechanisms of factor fibroblast growth factor inducible 14(Fn14)in the high glucose induced-cardiomyocyte hypertrophy. Method To observe the expression of collagenⅠ, connective tissue growth factor ( CTGF ) , transforming growth factor-β1 ( TGF-β1 ) , and Fn14 in high glucose induced-cardiomyocyte hypertrophy. Fn14 expressions was down-regulated by siRNA interference technique, and then the expressions of collagen Ⅰ, CTGF, and TGF-β1 were observed, and the mechanism was also explored. Results The expression of collagen I, CTGF and TGF-β1 was significantly up-regulated after high glucose induced-cardiomyocyte hypertrophy for 72 h. At the same time, the expression of Fn14 was increased after 72 h-treatment, and reached the peak at concentration of 30 mmol/L high glucose. High glucose could not up-regulated the expression of collagenⅠ, CTGF, and TGF-β1 after siFn14 interference, while the same result was observed in the expression of p-JNK. Conclusion The expressions of collagenⅠ, CTGF, TGF-β1, and Fn14 in cardiomyocyte of neonatal rats were induced by high glucose. While Fn14 expression was inhibited, the expressions of collagenⅠ, CTGF, and TGF-β1 were down-regulated, which seems to be involved with p-JNK signaling pathway.

Objective To compare the results of the anterior approach (AA) with the conventional approach in the treatment of large hepatocellular carcinoma (HCC).Methods We searched the Medline,PubMed,Cochrane Library,Wanfang database on randomized clinical controlled trials and non-randomized clinical controlled trials comparing AA with the CA in right hepatic resection for large hepatocellular carcinoma.The data were analyzed with the RevMan5 software.Results Five non-randomized clinical controlled trials (NRCTs) and three randomized clinical controlled trials involving 615 patients (304 in the AA group,311 in the CA group) were enrolled into the analysis.There was no significant difference in the operation time between the two groups.Compared with the CA,the AA had lower intraoperative blood loss (WMD=-680.2 ml; 95％CI,-1023.97～-336.43;P=0.0001),blood transfusion rate (OR=0.38;95％ CI,0.25～0.59;P＜0.0001),intraoperative tumor rupture (OR=0.33;95％CI,0.11～0.97;P=0.04),surgical complication (OR=0.59;95％CI,0.38 ～ 0.93 ; P =0.02),hospital mortality (OR =0.37 ; 95 ％ CI,0.21 ～ 0.67 ; P =0.0009),and hospital stay (WMD=-4.75 d;95％CI,-7.82～-1.67;P=0.002).Conclusion AA is superior to CA in the treatment of larger.The operation time is the same for the 2 approaches.

Objective To explore the effects of nerve growth factor (NGF) and low frequency electricity stimulus on the configurations of skeletal muscle cells and the change of muscle fibre types in the denervated skeletal muscle separately.Methods The denervated rat model was established and model animals were injected with the NGF and given the stimulus (frequency=2 Hz) about 30 days. The configurations and the change of muscle fibre types were observed by immunohistochemistry and image analysis.Results The muscle fibre was in chaos and the boundary was not obvious among cells in the denervated rats; the muscle fibre of the denervated rats with NGF injection and low frequency electricity stimulus was more regular and the boundary of cells was clearer, the cells number was more than those of the denervated rats. Compared to normal rats, the proportion of Ⅰ muscle fibre in the denervated rats increased ( P<0.05), whereas the proportion of Ⅱ muscle fibre decreased ( P<0.05); it had no significant differences of the two types of muscle fibre between the denervated rats with NGF injection, low frequency electricity stimulus and the denervated rats ( P<0.05).Conclusion NGF injection and low frequency electricity stimulus can make the configurations of denervated muscle to better.

Objective To investigate the efficacy and safety of indocyanine green fluorescence imaging in the lymph node dissection of radical thyroidectomy.Methods Radical thyroidectomy was performed using indocyanine green fluorescence imaging technology for two patients at the Department of General Surgery of Chinese People's Liberation Army (PLA) General Hospital in July 2018.Indocyanine green was injected into the thyroid glands after bilateral thyroid glands were exposed during operation.Bilateral total thyroidectomy plus central lymph node dissection was performed in case 1,and bilateral total thyroidectomy plus central area and left lateral area(area Ⅱ a,Ⅲ,Ⅳ) lymph node dissection was performed in case 2.Both operations were performed under the guidance of real-time fluorescence imaging system.The total number of lymph nodes detected,the number of small lymph nodes (diameter less than 3 mm),the level of parathyroid hormone(PTH),the incidence of complications such as hypocalcemia,hoarseness and short-term recurrence were observed.Results After excitation by the near-infrared light of the fluorescence detector probe,the display showed that the parathyroid gland and surrounding tissues were not visualized,and the thyroid glands and lymph nodes were brightly illuminated.The number of lymph nodes dissected in the central region of the two patients was 20 (13 with diameter less than 3 mm) and 10(6 with diameter less than 3 mm),respectively.For case 2,13 lymph nodes were dissected in the left lateral area (area Ⅱ a,Ⅲ,Ⅳ),and 8 lymph nodes with diameter less than 3 mm were dissected.There were no complications such as hypocalcemia and hoarseness after operation.The levels of parathyroid hormone and serum calcium were normal on the first day and 3 months after operation.There was no recurrence or metastasis of the tumors by ultrasonography 3 months after operation.Conclusion Indocyanine green fluorescence real-time imaging technology can help to identify lymph nodes specifically during radical thyroidectomy,and can achieve real-time dynamic imaging,which can make lymph node dissection more thorough and can be used as a new method for lymph node tracing in thyroid cancer surgery.

Objective:To study the expression of miR-496 in gastric cancer cells, and explore its role and mechanism in the proliferation and apoptosis of gastric cancer cells. Methods:Real-time fluorescence quantitative PCR (qPCR) was used to detect the expression of miR-496 in normal gastric epithelial cell lines and gastric cancer cell lines AGS and MKN45. miR-496 was knocked down in AGS cells with the lowest expression level, and a negative control group and a blank control group were set up. Cell proliferation and apoptosis were detected by CCK8 assay and flow cytometry. LYN, the target gene of miR-496, was screened using bioinformatics software, and the effect of transfection of miR-496 on LYN expression was detected by qPCR. Subsequently, rescure experiment was conducted to further study the mechanism of miR-496 on gastric cancer cells through regulation of LYN. Data were analyzed by GraphPad Prism 9 software. Measurement data were presented as mean ± standard deviation, and the comparison between the two groups was performed by t test. Results:The expression of miR-496 in AGS and MKN45 was significantly lower than that in normal gastric epithelial cells ( P<0.05). After overexpression of miR-496, the proliferation of AGS cells could be inhibited and the apoptosis ratio of AGS cells could be significantly increased ( P<0.05). QPCR results showed that miR-496 overexpression group could inhibit the expression of LYN ( P<0.05). Bioinformatics analysis showed that miR-496 binds to LYN kinase ( LYN) 3 ′UTR region, and overexpression of miR-496 can inhibit the expression of LYN in AGS cells, while CCK8 rescue experiment showed that overexpression of LYN could remove the inhibitory effect of miR-496 on cell proliferation. Flow cytometry showed that LYN expression could cancel the promoting effect of miR-496 on apoptosis ( P<0.05). Conclusion:miR-496 is low expressed in gastric cancer cells, and it inhibits the proliferation and promotes apoptosis of gastric cancer cells by targeting the expression of LYN in gastric cancer cells.

Objective:To study the risk factors of complications after bowel resection for acute mesenteric ischemic disease.Methods:Retrospective case-control study was used to analyze the case data of 68 patients diagnosed with acute mesenteric ischemic disease (AMI) with bowel resection at the First Medical Center of the PLA General Hospital from January 2010 to January 2020, including 43 males and 25 females. The patients were divided into complication group ( n=21) and the non-complication group ( n=47) according to whether they had complications after surgery. The risk factors associated with the development of postoperative complications were analyzed by multivariate Logistic stepwise regression method to determine the risk factors with clinical significance. Measurement data with normal distribution were expressed as mean ± standard deviation ( ± s), and t-test was used for comparison between groups. Chi-square test was used for comparison between count data groups. Results:Univariate analysis showed that age >60 years, Marshall score≥2, type of resected bowel, pathology suggestive of irreversible transmural necrosis, length of ICU stay >6 d, length of mechanical ventilation >2 d, American Society of Anesthesiologists (ASA) classification, and preoperative procalcitonin≥2 ng/mL were the risk factors affecting the development of complications after bowel resection for acute mesenteric ischemic disease risk factors ( P<0.05). Multivariate Logistic regression analysis showed that age>60 years ( HR=12.364, 95% CI: 1.135-134.662, P=0.039) and preoperative procalcitonin ≥2 ng/mL ( HR=14.144, 95% CI: 1.280-156.303, P=0.031) were independent risk factors for the development of postoperative complications after AMI parallel bowel resection. Conclusion:The rate of complications after combined bowel resection for AMI is high. When patients are combined with age>60 years and preoperative procalcitonin≥2 ng/mL, preoperative prevention of postoperative complications should be emphasized to improve the prognosis of patients.

In order to identify rat ovarian germ cells, we expressed and purified rat RVLG protein in Escherichia coli cells and prepared a mouse anti-rat RVLG polyclonal antibody. The rat RVLG cDNA was obtained from rat testicle tissue by RT-PCR and was cloned into the vector pMD19-T. Sequence analysis proves that the cloned RVLG cDNA fragment was 60 bp longer than that released in the GenBank (NM_001077647), resulting from an alternative splicing of the RVLG pre-mRNA. The RVLG cDNA was double digested with the restriction endonucleases BamH I and EcoR I, and then was extracted from gel and inserted into the prokaryotic expression vector pGEX-4T-1. The recombinant expression plasmid pGEX-RVLG was verified for successful construction and then was transformed into Escherichia coli BL21(DE3) for induction to express the GST-RVLG fusion protein by IPTG. The GST-RVLG fusion protein was expressed in Escherichia coli BL21 (DE3) at a high level which accounts for more than 10% of the total bacterial cellular protein. The purified RVLG protein was used as an antigen to immunize KM mouse for the production of polyclonal antibody in ascetic fluid followed by celiacly injecting the mouse with S180 cells. The mouse anti-rat RVLG antibody was analyzed by ELISA, Western blotting and immunohistochemistry for its specificity and titer. The antibody could recognize RVLG protein specifically and its titer was about 1:20 000. These results confirm that the mouse anti-rat RVLG polyclonal antibody with high affinity and specificity has been prepared successfully, and lay a foundation for our ongoing research on the specific expression of RVLG in rat ovary.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Base Sequence ; Cloning, Molecular ; DEAD-box RNA Helicases ; biosynthesis ; genetics ; immunology ; DNA, Complementary ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Female ; Mice ; Molecular Sequence Data ; Ovary ; cytology ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

Aim To explore the protective effects of LPPC ( procyanidins extracted from the litchi pericarp) on cardiomyocyte apoptosis in septic rats and its mech-anisms. Methods The rats were randomly divided in-to 5 groups, and were given orally the drug for two weeks continuously. The control group ( control) and sepsis model group ( LPS ) were given distilled water once a day. LPPC low, medium and high dose groups were given LPPC 50 , 100 , 200 mg · kg-1 · d-1 re-spectively which were prepared freshly every day. After the treatment, sepsis animal models were established. Except for the control group, other groups were injec-ted LPS (lipopolysacchride, 10mg·kg-1) intraperito-neally to induce acute sepsis model. 4hrs later, rat se-rum was collected, isoenzyme ( CK-MB ) , lactate de-hydrogenase ( LDH ) and activity of aspertate amin-otransferase ( AST/GOT) were detected. Then rat car-diac tissue was obtained and cardiac tissue malondial-dehyde ( MDA ) , total antioxidant capacity ( T-AOC ) and reduced glutathione ( GSH ) content were deter-mined. TUNEL staining was performed to analyze the apoptosis of myocardial cells. Cleaved caspase-3 and TNF alpha protein expressions were analyzed by West-ern blot. Results Compared with the control group ( control) , serum of sepsis model group rats CK-MB, LDH, AST/GOT and cardiac tissue MDA content were significantly increased (P<0. 01). At the same time, the activity of cardiac tissue T-AOC and GSH de-creased obviously ( P<0. 01 ) . The apoptotic myocar-dial cells increased significantly ( P<0. 01 ) , and the expression level of cleaved caspase-3 and TNF alpha decreased obviously ( P <0. 01 ) . LPPC pretreatment significantly decreased the serum CK-MB, LDH, AST/GOT and tissue MDA content, increased tissue T AOC and GSH activity, attenuated apoptosis of rat myocardi-al cells significantly, and decreased expression level of cleaved caspase-3 and TNF alpha. Conclusion LPPC pretreatment can significantly attenuate rat myocardial cell apoptosis induced by sepsis, and the underlying mechanisms may be related to its anti-oxidative effects.