Objective To investigate the mechanisms of metformin in inducing autophagy of gastric cancer MNK-45 cells.Methods Human gastric cancer MNK-45 cells in logarithmic growth phase were incubated in the culture plates,and were divided into the intervention group [gastric cancer MNK-45 cells were intervened by metformin at different concentrations (2,4,8,16,32,64 mmol/L) for 24,48,72 hours] and the control group (gastric cancer MNK-45 cells were cultured in the DMEM medium).The inhibition rate of gastric cancer MNK-45 cells was detected by MTT method.The IC50 value of metformin on gastric cancer MNK-45 cells was 17 mmol/L.Gastric cancer MNK-45 cells were intervened by metforrnin at 17 mmol/L for 48 hours in the experimental group.Gastric cancer MNK-45 cells in the control group were cultured in DMEM medium at 17 mmol/L for 48 hours.The apoptosis of the gastric cancer MNK-45 cells of the 2 groups were detected by flow cytometry.The mRNA expressions of Bax and Bcl-2 of the 2 groups were detected by RT-PCR.The protein expressions of type Ⅰ LC3b,type Ⅱ LC3b,beclinl,AKT,p-AKT,mTOR,p-mTOR,P70s6k,p-P70s6k of the 2 groups were detected by Western blot.The measurement data were presented as (x) s,and were analyzed using the one-way ANOVA or repeated measures ANOVA.Data of the 2 groups were compared using the t test.Results The inhibition rates of gastric cancer MNK-45 cells were 3.0％ ± 1.1％,8.6％ ± 1.7％,15.9％ ± 1.6％,26.1％ ± 3.4％,37.5％ ± 2.3％,49.7％± 3.6％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 24 hours,5.2％± 1.9％,10.4％±2.1％,26.9％± 1.6％,49.5％± 1.6％,59.1％±2.0％,82.1％±2.2％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 48 hours,and 9.5％ ± 2.2％,17.6％ ± 1.4％,30.6％ ± 2.6％,63.2％ ± 2.6％,78.9％ ± 1.4％,93.3％ ± 2.7％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 72 hours.There were significant differences in the inhibition rates among the 6 groups at the same time points (F =155.174,728.229,743.826,P ＜ 0.05),and significant differences were also observed within the same group at different time points (F =39.420,58.692,166.125,30.383,117.517,311.642,P ＜ 0.05).The apoptosis rates of gastric cancer MNK-45 cells in the experimental group and the control group were 25.4％ ± 1.7％ and 6.9％ ± 0.5％,with significant difference between the 2 groups (t =18.378,P ＜0.05).The relative mRNA expressions of Bax/Bcl-2 mRNA in the experimental group and the control group were 1.88 ± 0.16 and 1.00 ± 0.00,with significant difference between the 2 groups (t =9.743,P ＜ 0.05).The relative protein expressions of LC3 Ⅱ/LC3 Ⅰ and beclin 1 in the gastric cancer MNK-45 cells were 1.65 ± 0.08 and 1.47 ± 0.06 in the experimental group and 0.79 ± 0.03 and 0.56 ± 0.06 in the control group,with significant difference between the 2 groups (t =18.023,18.283,P ＜ 0.05).The relative protein expressions of AKT and P70s6k in the gastric cancer MNK-45 cells were 0.80 ±0.14 and 0.97 t0.21 in the experimental group and 0.96 ±0.17 and 1.37 ±0.23 in the control group,with no significant difference between the 2 groups (t =2.103,1.699,P ＞0.05).The relative protein expressions of mTOR and p-mTOR were 0.58 ± 0.l 1 and 0.57 ±0.15 in the experimental group and 1.88 ±0.23 and 2.36 ±0.25 in the control group,with significant difference between the 2 groups (t =11.293,10.979,P ＜ 0.05).No p-AKT and p-P70s6k expression was detected in the experimental group,and the expressions of p-AKT and p-P70s6k in the control group were 1.00 ± 0.00 and 1.00 ± 0.00,respectively.Conclusions Metformin could induce autophagy,inhibit proliferation and promote apoptosis of gastric cancer MNK-45 cells.The mechanism may be associated with the inhibition of mTOR expression and the expression of mTOR downstream proteins p-P70s6k by mefformin,and then the autophagy of gastric cancer MNK-45 cells happens.

Objective To explore the therapeutic efficacy of nanometer high-frequency square pulse light technology targeting kidney cancer.Methods Fifty BALB/c nude mice were vaccinated with human ACHN cell line and randomly divided into 1 control group and 4 therapeutic groups.The 4 therapeutic groups were cured with high-frequency square pulse light and nanometer high-frequency square pulse light.The treatment cycle was 4 weeks.The tumor growth condition and tumor-repres-sion change were observed and compared.Results The tumor volumes of the control group in-creased obviously,whereas the tumor volumes of the therapeutic groups decreased obviously or in-creased gently.The mean tumor volume and the tumor growth curve of the therapeutic groups were significantly lower than those of the control group(P＜0.05),but there was no significant difference between the therapeutic efficacy of the kidney cancer using high-frequency square pulse light and nanometer high-frequency square pulse light(P＞0.05).Synteresis of kidney carcinogenesis experiments results indicated that using high-frequency square pulse light and nanometer high-frequency square Dulse light could prevent the production and development of the kidney cancer(P＜0.05),but the svnteresis efficacy of the 2 methods had no obvious difference(P＞0.05).Conclusion Using highfrequency square pulse light and nanometer high-frequency square pulse light can cure the kidney cancer and,to some extent,prevent the production and development of kidney cancer.

Objective: To investigate the incidence of adverse events following immunization (AEFI) of human papillomavirus (HPV) vaccines in Hangzhou City from 2017 to 2021, so as to provide insights into safety monitoring and evaluation for HPV vaccines. Methods: The AEFI caused by immunization of bivalent (HPV2), quadrivalent (HPV4) and nonavalent HPV vaccines (HPV9) reported in Hangzhou City from 2017 to 2021 were captured from the AEFI Surveillance Module of Chinese Disease Control and Prevention Information System, and HPV vaccination data were captured from the Zhejiang Municipal Immunization Information Management System. The incidence, temporal distributions and clinical symptoms of AEFI were analyzed. Results: Totally 922 310 doses of HPV vaccines were immunized in Hangzhou City from 2017 to 2021, and 232 cases with AEFI were reported, with an overall incidence rate of 25.15/105 doses. The reported incidence rates of AEFI caused by HPV2, HPV4 and HPV9 vaccination were 31.13/105 doses, 25.93/105 doses and 22.01/105 doses, respectively. General reactions and abnormal reactions were predominant AEFI, and the reported incidence rates of general reactions and abnormal reactions were 21.58/105 doses and 2.60/105 dose, respectively. AEFI predominantly occurred 0 to 1 day post-immunization (165 cases, 71.12%), and the main clinical symptoms included local swelling of injection sites, hard tubercle and fever, with reported incidence rates of 10.30/105 doses, 5.96/105 doses and 6.18/105 doses, respectively. Conclusions Low incidence of AEFI was reported following HPV vaccination in Hangzhou City from 2017 to 2021, and all AEFI were mild. The safety of HPV2, HPV4 and HPV9 remains high.

Objective To systematically investigate the clinical efficiency and safety of ligation of inter sphincteric fistula tract versus incision-thread-drawing procedure for complicated anal fistula.Methods Searched PubMed,The Cochrane Library,CNKI,WanFang Data,and VIP from inception to May 2016,to collect randomized controlled trials of ligation of inter sphincteric fistula tract versus incision-thread-drawing procedure for complicated anal fistula.Search term included ligation of inter sphincteric fistula tract,fistula,incision-thread-drawing procedure,randomized controlled trial.The literatures were screened according to inclusive criteria,data were extracted and the quality of included studies was evaluated,and then meta-analysis was performed using RevMan 5.2 soft ware.A total of 5 randomized controlled trials including 305 patients were included.Results The results of meta-analysis showed that compared with incision-thread-drawing procedure,ligation of inter sphincteric fistula tract had a significant difference in amount of bleeding during surgery (MD =-18.30,95％ CI:-19.91 ～-16.69,P ＜ 0.000 01),the duration of pain (MD =-4.38,95％ CI:-4.69 ～-4.08,P ＜ 0.000 01),healing time (MD =-10.28,95％ CI:-15.71 ～-4.86,P =0.0002),hospital stay (MD =-7.44,95％CI:-10.87～-4.02,P＜0.000 1),recurrence rate (OR=0.31,95％CI:0.10～0.91,P=0.03).There was no significant difference in Operation time (MD =-5.83,95 ％ CI:-7.64 ～-4.02,P ＜ 0.000 01),effective percentage (OR =4.35,95％ CI:0.89 ～ 21.32,P =0.07) between both groups.Conclusion Compared with incision-thread-drawing procedure,ligation of inter sphincteric fistula tract shows significant advantage in cure rate,postoperative healing time,reducing post-operation pain,anal function protection and recurrence rate.

Objective:To compare the safety and immunogenicity of Sabin strain-based inactivated poliovirus vaccine (sIPV) and the liquid form of typeⅠ+ Ⅲ bivalent oral poliovirus vaccine (bOPV) administered to infants aged ≥2 months in different schedules.Methods:A randomized, blinded, single-center, parallel-group controlled trial was conducted in Hangzhou from 2017 to 2018. Healthy infants aged ≥2 months were enrolled and randomized to receive the vaccines on a schedule of 2, 3, 4 months. Group 1sIPV+ 2bOPV was given one dose of sIPV and two doses of bOPV; group 2sIPV+ 1bOPV was administrated two doses of sIPV and one dose of bOPV; group 3sIPV received three doses of sIPV. Adverse events (AEs) following vaccination were recorded. Blood samples were collected from the subjects (excluding the quitters or subjects against the trial plan) 28-35 d after the full-course immunization. A microneutralization assay was performed to detect the geometric mean titers (GMTs) of neutralizing antibodies against polio virus of Ⅰ, Ⅱ and Ⅲ types. The seroconversion rates of neutralizing antibodies were also calculated.Results:The overall incidence of AEs following vaccination was 3.57% in 1sIPV+ 2bOPV group, 3.61% in 2sIPV+ 1bOPV group and 1.19% in 3sIPV group (χ 2=1.190, P=0.552) and no severe AEs were reported. The antibody seroconversion rates in 1sIPV+ 2bOPV, 2sIPV+ 1bOPV and 3sIPV groups were respectively 100% (84/84), 100% (83/83) and 100% (84/84) against type Ⅰ poliovirus, 81% (68/84), 96% (80/83) and 99% (83/84) against type Ⅱ poliovirus(χ 2=21.469, P<0.001), and 100% (84/84), 100% (83/84) and 100% (84/84) against type Ⅲ poliovirus. In 1sIPV+ 2bOPV, 2sIPV+ 1bOPV and 3sIPV groups, the GMTs of antibody were 1 024.00, 1 015.48 and 982.61 against type Ⅰ poliovirus ( F=2.742, P=0.066), 16.81, 107.94 and 218.85 against type Ⅱ poliovirus ( F=33.570, P<0.001), and 990.75, 990.36 and 613.92 against type Ⅲ poliovirus ( F=37.886, P<0.001). Conclusions:sIPV and bOPV administered in different schedules showed good safety and immunogenicity in infants aged≥2 months. The GMT and the seroconversion rate of neutralizing antibody against type Ⅱ poliovirus after vaccination were higher in 2sIPV+ 1bOPV and 3sIPV group than in 1sIPV+ 2bOPV group. Higher GMT of neutralizing antibody against type Ⅲ poliovirus was induced in 1sIPV+ 2bOPV and 2sIPV+ 1bOPV groups than in 3sIPV group.

Objective:To investigate the effects of Qingfei Shenshi Decoction on the expressions of matrix metalloproteinase-2 (MMP-2) and MMP-9 and tissue inhibitor of metalloproteinase timps-1 (TIMP-1) in lung tissue of asthma mice.Methods:Totally 50 male BALB/C mice were divided into 5 groups: normal group, model group, dexamethasone group, Qingfei Shenshi Decoction low- and high-dosage groups (10 mice /group) according to random number table method. Asthma model mice were prepared by ovalbumin (OVA) challenge method. After successful modeling, the dexamethasone group was given dexamethasone for gavage at the rate of 1.56 mg/kg, while Qingfei Shenshi Decoction groups were given high and low doses of Qingfei Shenshi Decoction for gavage at the rate of 14.235 g/kg and 28.470 g/kg, respectively. Normal group and model group were given 0.9% sodium chloride solution by gavage. At the end of gavage administration for 4 weeks, the airway reactivity (Penh value) in each group was detected; HE staining was used to observe the pathological changes of lung tissue; the contents of interleukin-6 (IL-6), interleukin-1β (IL-1β) and tumor necrosis factor -α (TNF-α) in alveolar lavage fluid were determined by enzyme Linked immunosorbent assay (ELISA); the expressions of MMP-2, MMP-9 and TIMP-1 in lung tissue were detected by Western-blot.Results:Compared with model group, the damage of airway wall and alveolar wall of lung tissue in Qingfei Shenshi Decoction groups was significantly reduced. Compared with model group, the Penh value, IL-6, IL-1β and TNF-α levels in Qingfei Shenshi Decoction low- and high-dosage groups decreased ( P<0.05), and the expressions of MMP-9, MMP-2 and TIMP-1 in lung tissue decreased ( P<0.05), with a certain dose dependence. Conclusion:Qingfei Shenshi Decoction can effectively alleviate airway inflammation, reduce airway hyperresponsiveness, improve lung function and inhibit airway remodeling in asthmatic mice. Its mechanism may be related to down-regulating the expressions of MMP-2, MMP-9 and TIMP-1.

Objective:To investigate the correlation between serum complement C1q/tumor necrosis factor associated protein 3 (CTRP3) and carotid atherosclerosis in patients with type 2 diabetes mellitus (T2DM) and nonalcoholic fatty liver disease (NAFLD).Methods:From January 2018 to December 2019, 111 T2DM patients hospitalized in the Endocrinology Department of Nantong Third People ′s Hospital Affiliated to Nantong University, and 30 healthy physical examiners in the physical examination center of Nantong Third People 's Hospital Affiliated to Nantong University in the same period were selected. Thirty cases of healthy physical examination were the control group, 111 cases of T2DM were divided into 52 cases of T2DM group and 59 cases of T2DM+NAFLD group according to whether they were combined with NAFLD. The cross-sectional study method was used to collect the relevant clinical data of three groups. The comparison data between multiple groups conformed to the normal distribution and the variance was uniform. One way ANOVA was used. SNK- q test was used for pairwise comparison, χ2 test for qualitative data comparison. The correlation between carotid intima-media thickness (IMT) and influencing factors was analyzed by partial correlation analysis, and the influencing factors of carotid IMT were analyzed by multi factor linear regression. Results:In the control group, T2DM group and T2DM+NAFLD group, body mass index (BMI) (23.65±2.81), (25.52±3.12), (24.90±2.94) kg/m 2,systolic blood pressure (119.43±15.81), (130.63±10.20), (139.37±14.11) mmHg, diastolic blood pressure (72.93±9.74), (73.40±9.44), (77.97±10.00) mmHg, and fasting blood glucose (5.12±0.77), (9.78±1.37), (9.24±1.46) mmol/L,glycosylated hemoglobin (HbA1c) (4.87±1.43)%, (7.99±1.10)%, (8.56±1.29)%,homeostasis model assessment of insulin resistance (HOMA-IR)(1.56±0.37),(2.80±1.00), (3.47±0.94), high density lipoprotein cholesterol (HDL-C) (1.52±0.34),(1.23±0.31), (1.22±0.31) mmol/L,low density lipoprotein cholesterol (LDL-C) (2.41±0.53), (2.73±0.61), (2.93±0.59) mmol/L, CTRP3 (292.93±68.54), (241.69±61.01), (150.80±56.67) μg/L, the difference between groups were statistically significant ( F=3.712,23.023,4.074,134.285,90.818,47.105,10.139,7.941,60.035,all P<0.05). Pairwise comparison shows that the systolic blood pressure, diastolic blood pressure, HbA1c and HOMA-IR in T2DM+NAFLD group were higher than those in control group and T2DM group,and CTRP3 was lower than those in control group and T2DM group, the difference was statistically significant (all P<0.05). BMI, fasting blood glucose, HbA1c, HOMA-IR, HDL-C and LDL-C in T2DM group were higher than those in the control group, CTRP3 was lower than that in the control group (all P<0.05). In the control group, T2DM group and T2DM+NAFLD group, IMT were (0.75±0.13), (1.11±0.17) and (1.25±0.15) cm; Crouse scores were (1.28±0.97), (3.22±1.42) and (4.54±1.22); the plaque detection rates 16.7%(5/30), 65.4%(34/52) and 78.0%(46/59), and there were significant differences between the two groups ( F=105.941,67.063, χ2=32.108, all P<0.001). There were significant differences between the two groups (all P<0.05). T2DM+NAFLD group was the highest, followed by T2DM group, and the control group was the lowest. Partial correlation analysis showed that carotid IMT was positively correlated with systolic blood pressure, fasting blood glucose, HbA1c, HOMA-IR, triglyceride and LDL-C ( r=0.356, 0.572, 0.575, 0.620, 0.172, 0.291, all P<0.05), and negatively correlated with HDL-C and CTRP3 ( r=-0.335, -0.675, all P<0.001). Multivariate linear regression analysis showed that HbA1c, HDL-C and ctrp3 were the influencing factors of carotid atherosclerosis ( t=2.621, -3.764, -7.280, all P<0.05) Conclusion:Serum CTRP3 is associated with carotid atherosclerosis in T2DM patients with NAFLD,and may have a protective effect on vascular lesions in T2DM patients with NAFLD.

This study aims to propose a multifrequency time-difference algorithm using spectral constraints. Based on the knowledge of tissue spectrum in the imaging domain, the fraction model was used in conjunction with the finite element method (FEM) to approximate a conductivity distribution. Then a frequency independent parameter (volume or area fraction change) was reconstructed which made it possible to simultaneously employ multifrequency time-difference boundary voltage data and then reduce the degrees of freedom of the reconstruction problem. Furthermore, this will alleviate the illness of the EIT inverse problem and lead to a better reconstruction result. The numerical validation results suggested that the proposed time-difference fraction reconstruction algorithm behaved better than traditional damped least squares algorithm (DLS) especially in the noise suppression capability. Moreover, under the condition of low signal-to-noise ratio, the proposed algorithm had a more obvious advantage in reconstructions of targets shape and position. This algorithm provides an efficient way to simultaneously utilize multifrequency measurement data for time-difference EIT, and leads to a more accurate reconstruction result. It may show us a new direction for the development of time-difference EIT algorithms in the case that the tissue spectrums are known.

OBJECTIVE: To explore the role of mitochondrial permeability transition pore (mPTP) in mediating the protective effect of gastrodin against oxidative stress damage in H9c2 cardiac myocytes. METHODS: H9c2 cardiac myocytes were treated with HO, gastrodin, gastrodin+HO, cyclosporin A (CsA), or CsA+gas+HO group. MTT assay was used to detect the survival ratio of H9c2 cells, and flow cytometry with Annexin V-FITC/PI double staining was used to analyze the early apoptosis rate after the treatments. The concentration of ATP and level of reactive oxygen species (ROS) in the cells were detected using commercial kits. The mitochondrial membrane potential of the cells was detected with laser confocal microscopy. The expression of cytochrome C was detected with Western blotting, and the activity of caspase-3 was also assessed in the cells. RESULTS: Gastrodin pretreatment could prevent oxidative stress-induced reduction of mitochondrial membrane potential, and this effect was inhibited by the application of CsA. Gastrodin significantly lowered the levels of ROS and apoptosis-related factors in HO-exposed cells, and such effects were reversed by CsA. CsA significantly antagonized the protective effect of gastrodin against apoptosis in HO-exposed cells. CONCLUSIONS Gastrodin prevents oxidative stress-induced injury in H9c2 cells by inhibiting mPTP opening to reduce the cell apoptosis.
Adenosine Triphosphate ; analysis ; Apoptosis ; drug effects ; Benzyl Alcohols ; antagonists & inhibitors ; pharmacology ; Caspase 3 ; analysis ; Cell Line ; Cell Survival ; drug effects ; Cyclosporine ; pharmacology ; Cytochromes c ; analysis ; Glucosides ; antagonists & inhibitors ; pharmacology ; Humans ; Hydrogen Peroxide ; antagonists & inhibitors ; pharmacology ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondrial Membrane Transport Proteins ; physiology ; Myocytes, Cardiac ; drug effects ; metabolism ; Oxidative Stress ; Reactive Oxygen Species ; analysis

Deconvolution of potential drug targets of the central nervous system (CNS) is particularly challenging because of the complicated structure and function of the brain. Here, a spatiotemporally resolved metabolomics and isotope tracing strategy was proposed and demonstrated to be powerful for deconvoluting and localizing potential targets of CNS drugs by using ambient mass spectrometry imaging. This strategy can map various substances including exogenous drugs, isotopically labeled metabolites, and various types of endogenous metabolites in the brain tissue sections to illustrate their microregional distribution pattern in the brain and locate drug action-related metabolic nodes and pathways. The strategy revealed that the sedative-hypnotic drug candidate YZG-331 was prominently distributed in the pineal gland and entered the thalamus and hypothalamus in relatively small amounts, and can increase glutamate decarboxylase activity to elevate γ-aminobutyric acid (GABA) levels in the hypothalamus, agonize organic cation transporter 3 to release extracellular histamine into peripheral circulation. These findings emphasize the promising capability of spatiotemporally resolved metabolomics and isotope tracing to help elucidate the multiple targets and the mechanisms of action of CNS drugs.