Objectives:To study the methionine dependence(Met dependence) of human primary gastric cancer cells in vitro when Met in the culture medium was replaced by its precursor homocysteine(Hcy),and the effect of methionine starvation in combination with chemotherapeutical agents on gastric cancer cells. Methods:Fresh and sterile gastric cancer samples were managed to single cell suspensions and then were cultured in Met -Hcy + and Met +Hcy - medium separately,the proliferation of tumor cells in different culture media was examined by microcytotoxicity(MTT) assay.Meanwhile,the inhibition rate of tumor cells by ADM、DDP、5 FU、MMC and MTX in Met -Hcy +medium was separately tested. Results:①In Met -Hcy + medium,the human primary gastric cancer cell decreased;②Methionine deprivation in combination with chemotherapy enhanced obviously the killing capacity of each chemotherapeutical agent. Conclusions:①Human primary gastric cancer cells in vitro appears Met dependent.②The combined application of Met -Hcy + medium and different chemotherapeutical agents could enhance the antitumor effect of chemotherapy on primary gastric cancer cells.③MTT assay was an efficient way to examine the sensititivity of methionine starvation therapy combined with individualized chemotherapeutical agents.

Objectives:To study the methionine dependence (Met-dependence) of human primary gastric cancer cells in vitro when Met in the culture medicine was replaced by its precussor homocysteine(Hcy). Methods:Fresh and sterile gaotric cancer samples and normal gastric mucosa were managed into single-cell suspension and were cultured in the Met -Hcy + and Met\++ Hcy\+- medium separately.proliferation of these cells in the two different culture media was examined by cell counter and flow cytometry (FCM). Results:In the Met\++Hcy\+- medium,the growth of primary gastric cancer cells was inhibited,manifesting that the percentage of G 0G 1 tumor cells decreased and that of the S phase increased signiflcantly.However,normal gastric mucosal cells grew well and their cell cycles had no change. Conclusions:Human primary gastric cancer cells are Met dependent in vitro whereas normal gastric mucosal cells have no similar results.

Objective To investigate the effects of therapeutic exercise(cage running)on the expression of Bel-2 and Bax in neuronal cells after cerebral hemorrhage(ICH)in rats. Methods A total of 120 male Sprague-Dawley rats weighing 270 to 300 g were divided into a trial group(ICH and exercise group,n=40),a control group (ICH only,n=40)and a sham-operated group(sham ICH operation and no exercise,n=40).The brains were removed at 7,14,21 and 28 days after ICH.The activation of Bcl-2 and Bax WaS measured by immunohistochemical techniques,Western blotting and reverse transcriptase polymerase chain reaction(RT-PCR).Results (1)Bel-2-positive and Bax-positive cells appeared in tissues surrounding the hematoma in the cortex.The number of Bcl-2-posotive and Bax-positive cells was nearly zero in the sham-operated group.In the trim group.the number of Bcl-2-positive cells increased significantly from the 21 st to the 28th day.Bax.positive cells decreased from the 7th to the 28th day after ICH when compared with the control group and the sham-operated group.(2)The expression of Bcl-2mRNA and Bax-mRNA as shown by Western blotting and RT-PCR methods was synchronous with the immunohistochemistry results.The peaks of Bcl-2mRNA and BaxmRNA expression were a little earlier than those of Bcl-2 and Bax proteins. Conclusion Exercise can mitigate neuronal cell apoptosis by upregulating the expression of Bcl-2 and Bcl-2 mRNA,and downregulating the expression of Bax and Bax mRNA.

This study was aimed to compare the efficacy of gastric banding (GB), Roux en-Y gastric bypass (RYGBP) and biliopancreatic diversion (BPD) in the treatment of rats with type 2 diabetes mellitus (T2DM). Ani-mal models of T2DM were induced by streptozotocin (STZ) injection and high-sugar-fat diets. A total of 70 T2DM rats were randomly allocated into the GB group (G group, n = 20), RYGBP group (R group, n = 20), BPD group ( B group , n = 20 ) , and the sham operation group ( S group , n = 10 ) . The fasting blood glucose ( BG ) , triglyceride ( TG ) , total cholesterol ( TC ) and insulin ( INS ) content were determined before and 1 , 2 , 3 , 4 , 8 , 16 weeks after operation. The insulin sensitivity index (ISI) was calculated. The mortality and complications were ob-served in each group. The results showed that the fasting weight of the GB group, RYGBP group and BPD group were (324.4 ± 22.5) g, (338.9 ± 17.5) g, (333.3 ± 28.4) g, respectively. The BG content was (12.44 ± 1.29) mmol/L, (9.70 ± 0.81) mmol/L, (11.93 ± 2.39) mmol/L, respectively. The TC content was (2.32 ± 0.45) mmol/L, (2.22 ± 0.79) mmol/L, (2.13 ± 0.31) mmol/L, respectively. The TG content was (1.38 ± 0.32) mmol/L, (1.16± 0.41) mmol/L, (1.23 ± 0.35) mmol/L, respectively. The ISI were (-6.38 ± 0.29), (-6.67 ± 0.24), (-6.65 ±0.23), respectively. And the INS content of the RYGBP group were (69.43 ± 18.73) mU/L. There were signifi-cant differences between before and after operation on the 16th week ( P < 0 . 05 , P < 0 . 01 ) . The mortality rate was 5% in the GB group, 20% in the RYGBP group, and 35% in the BPD group. It was concluded that the GB, RYGBP and BPD are effective in reducing blood glucose and blood lipids in the treatment of rat with T2DM. The treatment effect is obvious in the improvement of insulin resistance ( IR ) .

Objective To establish the background information of physiological parameters for germ?free ( GF ) Taihu piglets. Methods In this study we selected 25 days old GF Taihu piglets and 4 conventional ( CV) littermates, the male and female ratio was 1∶3, to measure the normal clinical values of hematology and serum biochemistry, immunoglobu?lin concentration and main organ coefficients. The analysis of relative growths of main organ weight to body weight was con?ducted in the Taihu GF and CV pigs by allometric scaling model. Results (1) Twelve hematological parameters and 8 blood biochemical parameters in the GF piglets were significantly lower than those in CV pigs (P<0?05). (2) The aver?age body weight, IgM concentration of GF pigs and CV pigs had significant difference ( P <0?05 ) , and no mesenteric lymph nodes were found in the GF pigs. (3) The gut weight had the largest allometric association with body weight in the GF pigs, while spleen weight has the largest allometric association with body weight in the CV pigs. Both the weight of heart and stomach in CV and GF pigs had a negative allometric association with body weight (allometric coefficient b<1), respectively. Conclusions Different microbe control grades affect the body weight, hematology and serum biochemistry, expression of immunoglobulin and development of main organs in laboratory pigs.

Objective: To explore the feasibility of remotely obtaining complex information on traditional Chinese medicine (TCM) pulse conditions through voice signals. Methods: We used multi-label pulse conditions as the entry point and modeled and analyzed TCM pulse diagnosis by combining voice analysis and machine learning. Audio features were extracted from voice recordings in the TCM pulse condition dataset. The obtained features were combined with information from tongue and facial diagnoses. A multi-label pulse condition voice classification DNN model was built using 10-fold cross-validation, and the modeling methods were validated using publicly available datasets. Results: The analysis showed that the proposed method achieved an accuracy of 92.59% on the public dataset. The accuracies of the three single-label pulse manifestation models in the test set were 94.27%, 96.35%, and 95.39%. The absolute accuracy of the multi-label model was 92.74%. Conclusion Voice data analysis may serve as a remote adjunct to the TCM diagnostic method for pulse condition assessment.

Objective A lentivirus-mediated colon cancer cell line stably overexpressing human SAMHD1 gene was constructed to observe the effect of this gene on the ability of the cells to resist herpes simplex virus type 1 (HSV-1) infection. Methods p CDH-EF1-MCS-SAMHD1-T2 A-GFP recombinant plasmid was constructed using pCDH-EF1-MCS-T2 A-copGFP lentiviral vector. After confirming successful synthesis with sequencing, the recombinant plasmid and lentivirus packaging plasmids were co-transfected into 293 T cells. The pseudovirus solution was collected and concentrated, then human colon cancer cells were infected with the concentrated solution. The cell line overexpressing SAMHD1 gene was infected with HSV-1 in contrast to the control group, and the virus infection efficiency was assessed by Western blotting and immunofluorescence analyses. Results The pCDH-EF1-MCS-SAMHD1-T2 A-GFP recombinant plasmid was successfully constructed and verified by gene sequencing. Western blotting confirmed the overexpression of the SAMHD1 gene with higher levels of the gene in the transfected cells in contrast to control human colon cancer cell line. Western blotting and immunofluorescence showed that the cell line overexpressing human SAMHD1 gene could effectively inhibit the infection of HSV-1. Conclusion Lentivirus-mediated stable cell line overexpressing human SAMHD1 gene was effective in inhibiting HSV-1 replication and could help us to further investigate the function of SAMHD1.

Objective:To study the effects of Zuogui Pills on the expressions of miR-133b-3p and RhoA in osteoclasts of postmenopausal osteoporosis rats; To discuss its potential mechanism.Methods:SD female rats were randomly divided into normal group, model group, sham-operation group, and Zuogui Pills group using a random number table method, with 6 rats in each group. The model group and Zuogui Pills group were treated with oophorectomy to construct a rat model of osteoporosis. Zuogui Pills group was orally administered with Zuogui Pills decoction at a concentration of 10 g/kg for 12 consecutive weeks. Colorimetric method was used to measure the serum calcium and phosphorus levels of rats, and ELISA method was used to detect ALP levels. Bone density meter was used to measure the bone density of the femurs of rats in each group. The osteoclast of each group were cultured, and the expressions of RANKL and RUNX2 protein were detected by Western blot. MiRNA sequencing and differential expression analysis were performed on bone tissues of rats. Osteoclasts were treated with miR-133b-3p mimic and its negative control. The cell proliferation activity of osteoclasts was detected by cell counting kit-8 (CCK-8). The osteoclast differentiation activity was detected by the tartrate-resistant acid phosphatase staining. The dual-luciferase reporter assay was used to detect the relationship between miR-133b-3p and RhoA. The "rescue" experiment of miR-133b-3p mimic and RhoA co-expression were used to study the molecular regulatory mechanism of Zuogui Pills on osteoclast activity.Results:Compared with the model group, the bone mineral density of Zuogui Pills group significantly increased ( P<0.05, P<0.01), the levels of calcium and phosphorus in serum increased, the level of alkaline phosphatase ALP decreased ( P<0.05), the expression of RANKL protein decreased, and the expression of RUNX2 protein increased. Sequencing results showed that rno-miR-133b-3p was down-regulated in osteoclasts of postmenopausa osteoporosis rats treated with Zuogui Pills with the maximum difference ( P<0.01). Q-PCR results showed that the expression of miR-133b-3p in osteoclasts of Zuogui Pills group was significantly lower than that of the model group. The upregulation of miR-133b-3p could significantly promote the cell proliferation and differentiation of osteoclasts. RhoA overexpression could reverse the excessive proliferation and differentiation of osteoclasts caused by miR-133b-3p overexpression. Conclusions:RhoA is the target gene regulated by miR-133b-3p. Zuogui Pills can inhibit the activity of osteoclasts by regulating miR-133b-3p/RhoA axis, relieving the symptoms of osteoporosis.

Objective:To explore the relationship between perioperative heparin dose and postoperative bleeding.Methods:From January 2000 to October 2022, we searched PubMed, EMbase, Cochrane Library, CNKI, CBM, WanFang Data and other databases by computer. This study explored the literature of controlled clinical trials on the correlation between heparin dosage and postoperative bleeding, and used Review Manager 5.4 software for statistical analysis.Results:A total of 10 studies with a total of 829 patients were included in this study. The results of meta-analysis showed that postoperative blood loss in high-dose heparin patients was significant, and the difference was statistically significant between high-dose and low-dose groups( MD=108.4, 95% CI: 85.55-131.33, I2=0, P<0.0001). The proportion of blood transfusion was also significantly increased in the high-dose group( OR=2.32, 95% CI: 1.10-4.89, I2=62%, P=0.03). However, there was no effect on postoperative death at 30 days and ICU stay time. Conclusion:High dose heparin significantly increases postoperative blood loss and the proportion of postoperative transfusions required, but has no significant effect on postoperative death at 30 days and ICU stay time.

Objective To evaluate the acute exacerbation of chronic obstructive pulmonary disease(COPD)(AECOPD)status via combining clinical data,lung function parameters with CT radiomic features based on machine learning method.Methods A total of 343 COPD patients,including 158 AECOPD patients and 185 non-AECOPD patients were retrospectively selected and randomly divided into training and testing sets at a ratio of 7∶3.The radiomics features were calculated after automatically delineating the whole lung volume of interest(VOI).Five machine learning methods were used to construct the AECOPD diagnostic model,then the corresponding Radiomics score(Rad-score)was calculated in the training set and was validated in the testing set.The logistic-combined model was established after integrating age,Global Initiative for Chronic Obstructive Lung Disease(GOLD)classification,vital capacity(VC),forced vital capacity(FVC),forced expiratory volume in one second(FEV1),FEV1％pred,FEV1/FVC％,peak expiratory flow(PEF),maximum ventilatory volume(MVV),and Rad-score value.The area under the curve(AUC)of receiver operating characteristic(ROC)curve was calculated to evaluate the evaluated performance of all models.Results The logistic regression model had the best diagnostic performance,with AUC of 0.724 and 0.758 in the training and testing sets,respectively.The performance of the logistic-combined model to diagnose AECOPD was superior to that of the single logistic regression model,with the AUC of 0.777 and 0.760 in the training and testing sets,respectively.Conclusion A combination strategy including clinical data,lung function parameters,and CT radiomics may be helpful to diagnose AECOPD status,with moderate diagnostic performance.