OBJECTIVE To investigate the inhibitory effect and the possible mechanism of hydrogen sulfide （H2S） on the proliferation of cardiac fibroblasts. METHODS The heart of neonatal SD rats was collected， and cardiac fibroblasts were separated with differential centrifugation. Using sodium hydrosulfide as the donor of H2S， the effects of H2S on the proliferation of cardiac fibroblasts induced by angiotensin Ⅱ （Ang Ⅱ）， hydroxyproline content and the expression of sirtuin 3 （SIRT3） protein were detected. After SIRT3 knockdown with siRNA technology， the effects of H2S on the proliferation of cardiac fibroblasts induced by Ang Ⅱ， hydroxyproline content， the expressions of collagen Ⅰ （Col Ⅰ）， collagen Ⅲ （Col Ⅲ ） and optic atrophy protein 1 （OPA1） were detected. RESULTS H2S could inhibit the proliferation of Ang Ⅱ-induced cardiac fibroblasts， reduce the content of hydroxyproline and increase the expression of SIRT3 （P＜0.05）. After down-regulating the expression of SIRT3 with siRNA technology， the inhibition of H2S on the proliferation of Ang Ⅱ-induced cardiac fibroblasts and the reduction of hydroxyproline content were both inhibited， and the effect of H2S on reducing the expression of Col Ⅰ and Col Ⅲ and enhancing the expression of OPA1 was also significantly weakened. CONCLUSIONS H2S inhibits the proliferation of Ang Ⅱ -induced cardiac fibroblasts through increasing the expression of SIRT3.

Background::The complete mesogastrium excision (CME) based on D2 radical gastrectomy is believed to significantly reduce the local-regional recurrence compared with D2 radical gastrectomy in advanced gastric cancer, and it is widely used in China. This study aimed to explore whether D2 + CME is superior to D2 on surgical outcomes during gastrectomy from Chinese data.Methods::Feasible studies comparing the D2 + CME (D2 + CME group) and D2 (D2 group) published up to March 2020 are searched from electronic databases. The data showing surgical and complication outcomes are extracted to be pooled and analyzed.Results::Fourteen records including 1352 patients were included. The D2 + CME group had a shorter mean operative time (weighted mean difference [WMD] = -16.72 min, 95% confidence interval [CI]: -26.56 to - 6.87 min, P < 0.001), lower mean blood loss (WMD = -39.08 mL, 95% CI: -49.94 to -28.21 mL, P < 0.001), higher mean number of retrieved lymph nodes (WMD = 2.13, 95% CI: 0.58-3.67, P = 0.007), shorter time to first flatus (WMD =-0.31 d, 95% CI: -0.53 to - 0.10 d, P = 0.005), and postoperative hospital days (WMD =-1.09, 95% CI: -1.92 to -0.25, P = 0.010) than the D2 group. Subgroup analysis suggested that the advantages from the D2 + CME group were obvious in traditional open radical gastrectomy, proximal gastrectomy, and distal gastrectomy compared with D2 group. The evaluations of post-operative complications showed that the patients who underwent D2 + CME had a lower incidence of post-operative complications than the patients who underwent D2 surgery alone (relative risk [RR] = 0.65, 95% CI: 0.45-0.87, P = 0.003). The D2 radical gastrectomy plus CME improved 3-year overall survival (OS) (RR = 1.16, 95% CI: 1.02-1.32, P = 0.020) and lowered the local recurrence rate (RR = 0.51, 95% CI: 0.28-0.94, P = 0.030). The patients undergoing laparoscopic surgery or total gastrectomy had more significant advantages compared between D2 + CME and D2 groups in 3-year OS. Conclusion::The data from China show that D2 radical gastrectomy plus CME are reliable procedures and safety compared to D2 radical gastrectomy with faster recovery, lower risk, and better prognosis.

Objective:To investigate the application value of esophagogastric anastomosis with double muscle flap plasty in total laparoscopic radical resection of proximal gastric cancer.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 5 patients undergoing radical resection of proximal gastric cancer in Cancer Hospital of China Medical University from January to December 2020 were collected. All 5 cases were male, aged from 57 to 72 years, with a median age of 65 years. All 5 patients underwent total laparoscopic radical resection of proximal gastric cancer combined with esophagogastric anastomosis with double muscle flap plasty. Observation indicators: (1) operative situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted using outpatient examination and telephone interview to detect postoperative anastomosis, esophageal reflux, nutritional status, quality of life, tumor recurrence and metastasis of patients up to February 2021. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Operative situations: all 5 patients underwent surgery successfully. The operation time, time of esophagogastric anastomosis with double muscle flap plasty, number of lymph node dissected, volume of intraoperative blood loss, length of surgical incision of 5 patients were (316±41)minutes,(109±11)minutes, 26±4, (48±12)mL, (3.4±0.4)cm, respectively. Results of intraoperative rapid frozen section pathological examination showed negative of esophageal margin. (2) Postoperative situations: the time to postoperative initial flatus, time to postoperative initial food intake, during of postoperative hospital stay, cost of treat-ment of 5 patients were (4.8±1.5)days, (5.8±1.5)days, (11.6±2.1)days and (5.5±0.4)×10 4 yuan, respectively. Results of postoperative pathological examination of 5 patients showed gastric adeno-carcinoma in all 5 patients including 4 cases with moderately and poorly differentiated adeno-carcinoma and 1 case with highly differentiated adenocarcinoma, with the TNM staging of pT1a-3N0-1 M0 stage. Of the 5 patients, 1 case underwent postoperative mild pneumonia and was cured by conservative treatment such as anti-infection and promotion of sputum evacuation. (3) Follow-up: all 5 patients were followed up for 2 to 12 months, with a median follow-up time of 6 months. Of the 5 patients, 4 cases underwent anastomotic patency and 1 case underwent mild anastomotic stenosis who was improved after endoscopic treatment. None of the 5 patients underwent reflux esophagitis. The body mass index, the score of nutritional risk screening 2002, the score of patient-generated subjective global assessment and the score of tumor patient quality of life of 5 patients were 21 kg/m 2(range, 19-27 kg/m 2), 2(range, 1-2), 2(range, 1-3) and 47(range, 42-52), respectively. None of the 5 patients underwent tumor recurrence or metastasis. Conclusion:Esophagogastric anas-tomosis with double muscle flap plasty can be used in total laparoscopic radical resection of proximal gastric cancer which will lead to satisfactory short-term efficacy.

Tyrosine phosphatase SHP2 is a promising drug target in cancer immunotherapy due to its bidirectional role in both tumor growth promotion and T-cell inactivation. Its allosteric inhibitor SHP099 is known to inhibit cancer cell growth both and . However, whether SHP099-mediated SHP2 inhibition retards tumor growth anti-tumor immunity remains elusive. To address this, a CT-26 colon cancer xenograft model was established in mice since this cell line is insensitive to SHP099. Consequently, SHP099 minimally affected CT-26 tumor growth in immuno-deficient nude mice, but significantly decreased the tumor burden in CT-26 tumor-bearing mice with intact immune system. SHP099 augmented anti-tumor immunity, as shown by the elevated proportion of CD8IFN- T cells and the upregulation of cytotoxic T-cell related genes including , which decreased the tumor load. In addition, tumor growth in mice with SHP2-deficient T-cells was markedly slowed down because of enhanced anti-tumor responses. Finally, the combination of SHP099 and anti-PD-1 antibody showed a higher therapeutic efficacy than either monotherapy in controlling tumor growth in two colon cancer xenograft models, indicating that these agents complement each other. Our study suggests that SHP2 inhibitor SHP099 is a promising candidate drug for cancer immunotherapy.

Objective To investigate the regulation effects of Ngn2 gene transfection on retinal neuron differnetion in three-dimentional optic vesicle (OV) of mice.Methods OV was cultured in vitro using mouse induced pluripotent stem cells (iPSC) under specific conditions.During OV culture,it was transfected multiple times by lentivirus-mediated Ngn2 gene and then it was induced after maturation.The cells were specificly differentiated toward retinal nerve cells in OV.Using the green fluorescent protein (EGFP) gene as control,the differentiation of retinal nerve cells in OV was detected by immunohistochemistry.Reverse transcription PCR and Western blot were used to quantitatively detect the expressions of retinal neuron-specific proteins Pax6,Islet1 and Brn3b.Results The mouse iPS-derived OV was successfully cultured.The number of neural cells in the OV transfected with the Ngn2 gene was increased by the lentiviral-mediated lentivirus.The expressions of PAX6,Islet1 and Brn3b in the Ngn2 transfection group were significantly higher at the gene and protein levels than those in the control group,with significant differences between the two groups (P＜0.05).Conclusions The Ngn2 gene can effectively increase the number of retinal neuron differentiation in OV and make in vitro cultured OV more mature and form a more perfect retinal cell neural circuit.

Objective: To investigate the clinical value of endoscopic ultrasonography (EUS) and Multi-slice Spiral CT (MSCT) in the preoperativestaging of tumor(T) and lymph node (N) metastasis in patients with SiewertⅡand Ⅲ typeadenocarcinoma of esophagogastric junction(AEG). Methods: Clinical data of 145 Siewert Ⅱ and Ⅲ type AEG patientswithout preoperative chemoradiotherapy were retrospectively reviewed. Theyall received preoperative EUS and MSCT examination and underwent surgical resection, and the results of EUS and MSCT were compared with their postoperative pathologic staging. Results: The sensitivity, specificity, and accuracy of EUS for T stage in Siewert Ⅱ and Ⅲ type AEG were higher than those of MSCT. The total accuracy of EUS and MSCT were 90.3% and 63.5%, respectively, and the difference was statistically significant (χ²=29.52, P<0.01). The sensitivity of EUS for T1, T2 and T3 were 89.5%, 91.1% and 85.2%, respectively, which were significantly higher than 42.1%, 66.7% and 29.6% of MSCT (χ²=9.47, P<0.01 for T1; χ²=8.07, P<0.01 for T2; χ²=17.40, P<0.01 for T3). In addition, the total accuracy of EUS and MSCT for lymph node metastasis status of Siewert Ⅱ and Ⅲ type AEG were 75.9% and 64.8%, respectively, showing a statistically significant difference(χ²=4.23, P=0.04). The sensitivity of EUS for N1 and N2 were 82.1% and 79.2%, respectively, which were significantly higher than 53.6% and 60.4% of MSCT (χ²=5.24, P=0.02; χ²=4.48, P=0.03). There was no statistical significance for sensitivity of EUS and MSCT in N0 and N3 (P>0.05). Conclusion EUS diagnosis of T and N staging in Siewert Ⅱ/Ⅲ type AEG showed significantly greater performance than MSCT.

Objective To observe the clinical effects of arthroscopic reconstruction for anterior cruciate ligament ( ACL ) tear using tension-relieving technique and enhanced recovery after surgery ( ERAS ) . Methods Between May 2014 and June 2016, 80 patients with ACL tear were randomly di-vided into 2 equal groups. The experimental group was treated with arthroscopic reconstruction using hamstring tendon autograft and tension-relieving technique followed by ERAS; the control group was treated with arthroscopic reconstruction using hamstring tendon autograft only followed by conventional postoperative re-covery. The 2 groups were compared in terms of knee flexion angles at postoperative 2nd, 4th and 8th weeks, and the IKDC ( International Knee Documentation Committee ) , HSS ( Hospital for Special Surgery ) , and Lysholm scores at postoperative 3rd, 6th and 12th months. Results The 80 patients obtained an average follow-up of 12. 3 ± 1. 7 months. Arthroscopy one year postoperation revealed fine ACL growth without laxity or other complications in the experimental group. Joint stiffness appeared in one case at 2 months postoperation and ACL laxity was observed in 3 cases by arthroscopy at 12 months postoperation in the control group. The knee flexion angles at postoperative 2nd, 4th and 8th weeks, and the IKDC, HSS and Lysholm scores at postoperative 3rd, 6th and 12th months in the experimental group were significantly better than those in the control group ( P <0. 05 ) . In both groups, the IKDC, HSS and Lysholm scores at postoperative 3rd, 6th and 12th months and the knee flexion angles at postoperative 2nd, 4th and 8th weeks were all significantly im-proved than the preoperative values ( P <0. 05 ) . Conclusion In arthroscopic reconstruction for ACL tear, tension-relieving technique combined with ERAS can promote functional recovery of the knee and reduce postoperative complications, facilitating early recovery of sports function.

Objective To study the protective effect of Rhein on the kidney of type 2 diabetic rats induced by high fat diet.Methods A rat model of type 2 diabetes was induced by high fat diet combined with low dose streptozotocin 35 mg/kg.The diabetic rats were randomly divided into diabetes group, Low, middle and high rhein dose groups (50,100,150 mg/kg), metformin group (300 mg/kg) and normal control group.Blood glucose and urine micro albumin were measured at 0, 2, 4 and 8 weeks respectively.Serum creatinine, urea nitrogen, total cholesterol and triglyceride were measured at 8 weeks.HE staining was used to observe the pathological changes of renal tissue.Effects of rhein on the expression of transforming growth factor-β1 and Smad3 protein in renal tissue of diabetic rats were detected with Western Blot.Results The blood glucose and urine micro albumin in model group were significantly higher than those in normal control group.Each rhein dose group exhibited reduced blood glucose and urinary micro albumin in diabetic rats.The high rhein dose group showed significant reduction of blood glucose and urine micro albumin in diabetic rats (P<0.05).Compared with model group, rhein reduced the serum Cr, BUN, TC and TG values in each dose group, most significantly in the high rhein dose group (P<0.05).The obvious pathological changes of renal tissue in model group were observed with most improved changes in the high rhein dose group.The expression of TGF-β1 and Smad3 protein in renal tissue of diabetic rats decreased significantly (P<0.05).Conclusion Rhein has preventive effect on diabetic nephropathy.The mechanism may relate to the improvement of renal function, regulation of blood lipid and down regulation of TGF-β1 and Smad3 protein expression in renal tissue.

BACKGROUND: Beta-tricalcium phosphate (β-TCP)/alpha-calcium sulfate hemihydrate (α-CSH) artificial composite bone has a porous morphology and good biocompatibility, and it is helpful to improve the fusion rate in a spinal fusion model, which however has not yet been confirmed.OBJECTIVE: To investigate the preparation methods, biocompatibility and application effect of β-TCP/α-CSH composite bone in the spinal fusion model.METHODS: (1) Calcium sulfate dihydrate under certain conditions and at a proper temperature can be dehydrated to prepare α-CSH. Healthy bovine cancellous bone was decellularized, degreased and sintered under the certain condition and at the certain temperature to prepare β-TCP particles. Then, the β-TCP particles were dissolved in anhydrous ethanol, suspended, dried, and then used to prepare the β-TCP/α-CSH composite bone. Osteoblasts from the rabbit periosteum were co-cultured with the composite bone, and then cell morphology, adhesion and proliferation were observed. (2) Twenty New Zealand white rabbits were selected to make bilaterally posterolateral spinal fusion models of the multiple thoracic vertebrae, in which β-TCP/α-CSH composite bone was implanted into the left side (experimental group) and autogenous bone implanted into the right side (control group). The spinal fusion rate was compared between the two groups.RESULTS AND CONCLUSION: (1) Under the phase contrast microscope, a relatively small amount of L929 cells adhered to the composite bone after 3 days of co-culture, while the number of adherent cells became relatively dense. Under the scanning electron microscope, there were many crystalline particles on the surface of the composite bone,indicating a higher number of adherent cells on the composite bone surface. (2) The spinal fusion rate was increased at 4 weeks after implantation of β-TCP/α-CSH composite bone, which was significantly higher than that after implantation of autogenous bone (P < 0.05). (3) At 4 weeks after autogenous bone implantation, the bone trabecular bone was scarce and clumped, and newborn bone tissues and the boneless autograft bone were dominant. After 8 weeks after spinal fusion, the newborn bone tissues around the autograft were further increased. At 4 weeks after β-TCP/α-CSH composite bone implantation, there was no degradation of debris, but existed a few new bone tissues; at 8 weeks after implantation,the composite bone was surrounded by newborn bone tissues, and thickened trabecular bone and degradation of the composite bone were found. To conclude, the prepared β-TCP/α-CSH composite bone can achieve a higher fusion rate in the spinal fusion model.

Aim To discuss the effects and mechanism of Ganoderma lucidum polysaccharides and metformin on myocardial structure and hemodynamics in type 2 diabetic rats.Methods High fat diet combined with intraperitoneal injection of low dose streptozotocin 30 mg· kg -1 was applied to establish rat model of type 2 diabetes mellitus .The diabetic rats were randomly into normal control group ,diabetes group , ganoderma lucid-um polysaccharides group (600 mg· kg -1 ) , metformin group ( 600 mg · kg -1 ) , combination group ( ganoder-ma lucidum polysaccharides 300 mg · kg -1 +metform-in 300 mg· kg -1 ) .After 12 weeks′treatment,the lev-els of fasting serum glucose were determined and the hemodynamic parameters (LVSP,LVEDP,dp/dtmax,-dp/dtmax ) were determined.Collagen volume fraction ( CVF ) was detected by Van Gieson . Immunohisto-chemical method and Western blot were used to detect myocardial tissue MMP-2 protein expression .Results The fasting blood glucose was significantly decreased in the combined treatment group .Combined medication could significantly improve hemodynamic parameters in diabetic rats: reduced LVEP and raised LVEDP , dp/dtmax and -dp/dtmax .CVF was significantly decreased in combination group .The expression of MMP-2 in my-ocardial tissue was significantly inhibited .Conclusions The combination of Ganoderma lucidum polysaccha-ride and metformin can significantly improve the hemo-dynamic parameters in type 2 diabetic rats, and have a preventive effect on diabetic cardiomyopathy . The mechanism may be related to the down regulation of the expression of MMP-2.