Objective:To investigate the clinical outcomes of patients with multiple morphological abnormalities of the flagella (MMAF) caused by CFAP43 or CFAP44 gene mutations following intracytoplasmic sperm injection (ICSI). Methods:Clinical data and genetic information were retrospectively analyzed for 121 MMAF patients who attended Reproductive Medicine Center, Department of Obstetrics and Gynecology, the First Affiliated Hospital of Anhui Medical University during September 2014 to July 2020. Totally 9 MMAF patients were identified to carry CFAP43 or CFAP44 mutations, 5 of them (P3, P5, P7, P8, and P9) received ICSI treatments, the ICSI outcomes were further analyzed. Results:Sanger sequencing validated 9 MMAF patients harboring CFAP43 or CFAP44 biallelic mutations, our study firstly identified a novel homozygous mutation of CFAP43(c.4132delC: p.Arg1378Glufs*10), novel compound heterozygous mutations of CFAP43 (c.3938G>A: p.Arg1313Gln;c.4342G>A:p.Glu1448Lys) and novel compound heterozygous mutations of CFAP44 (c.1718C>A:p.Pro573His; c.4075G>A: p.Glu1359Lys). The 5 MMAF patients underwent 5 ICSI cycles, 4 healthy offspring were obtained. The rate of fertilization of CFAP43- or CFAP44-mutated MMAF patients following ICSI was 76.47% (39/51), 3 patients' wife got clinical pregnancy, 3 patients got live birth delivery, respectively. No significant differences were found in ICSI outcomes among CFAP43-mutated or CFAP44-mutated MMAF patients, DNAH1-mutated MMAF patients, and severe oligoasthenozoospermia group (all P>0.05) .Conclusion:CFAP43 or CFAP44 mutations are responsible for the malformation of sperm flagella and decrease of sperm motility, and validated as the important genetic causes of MMAF. CFAP43- or CFAP44-mutated MMAF patients could have a favorable treatment outcome following ICSI.

Objective:To investigate the clinical outcomes of patients with multiple morphological abnormalities of the flagella (MMAF) caused by CFAP43 or CFAP44 gene mutations following intracytoplasmic sperm injection (ICSI). Methods:Clinical data and genetic information were retrospectively analyzed for 121 MMAF patients who attended Reproductive Medicine Center, Department of Obstetrics and Gynecology, the First Affiliated Hospital of Anhui Medical University during September 2014 to July 2020. Totally 9 MMAF patients were identified to carry CFAP43 or CFAP44 mutations, 5 of them (P3, P5, P7, P8, and P9) received ICSI treatments, the ICSI outcomes were further analyzed. Results:Sanger sequencing validated 9 MMAF patients harboring CFAP43 or CFAP44 biallelic mutations, our study firstly identified a novel homozygous mutation of CFAP43(c.4132delC: p.Arg1378Glufs*10), novel compound heterozygous mutations of CFAP43 (c.3938G>A: p.Arg1313Gln;c.4342G>A:p.Glu1448Lys) and novel compound heterozygous mutations of CFAP44 (c.1718C>A:p.Pro573His; c.4075G>A: p.Glu1359Lys). The 5 MMAF patients underwent 5 ICSI cycles, 4 healthy offspring were obtained. The rate of fertilization of CFAP43- or CFAP44-mutated MMAF patients following ICSI was 76.47% (39/51), 3 patients' wife got clinical pregnancy, 3 patients got live birth delivery, respectively. No significant differences were found in ICSI outcomes among CFAP43-mutated or CFAP44-mutated MMAF patients, DNAH1-mutated MMAF patients, and severe oligoasthenozoospermia group (all P>0.05) .Conclusion:CFAP43 or CFAP44 mutations are responsible for the malformation of sperm flagella and decrease of sperm motility, and validated as the important genetic causes of MMAF. CFAP43- or CFAP44-mutated MMAF patients could have a favorable treatment outcome following ICSI.

Objective To estimate the dose-response relationship between sedentary behavior with mortality in patients with type 2 diabetes. Methods A total of 17786 type 2 diabetic patients were recruited as participants, who were included in National Basic Public Health Service in Changshu County of Suzhou City, Qinghe District and Huai'an District in Huai'an City of Jiangsu Province. Cox proportional hazards regression model and restricted cubic spline model were employed to estimate the dose-response relationship between sedentary behavior with all-cause and cause specific mortality in patients with type 2 diabetes. Results Among 78114.34 person-years of the fo1low-up, the median of follow-up time was 4 years, and 1285 deaths occurred during that period. Compared to patients with sedentary behavior≤2 h/d, the multivariate adjusted hazard ratios of all-cause death associated with sedentary behavior levels of 3-4 h/d, 5-6 h/d, and≥7 h/d were 1.05(95％CI 0.92-1.20), 1.20(95％CI 1.03-1.42), and 1.39 (95％CI 1.16-1.65), respectively. Eevry increase of 1 h/d in sedentary behavior was associated with an increased hazard of death from cardiovascular disease(CVD) of 4％(HR=1.04, 95％CI 1.01-1.07) and from other causes of 6％( HR=1.06, 95％CI 1.03-1.09) . However, no significant association between sedentary behavior and malignant tumor death was found. The multivariable restrictive cubic spline regression indicated that the linear dose-response relationships were found between sedentary time with the all-cause, CVD cause, and other cause of mortality ( Non-linear test, P>0.05) . Conclusion Longer sedentary behavior could increase the risk of mortality in patients with type 2 diabetes.

OBJECTIVETo observe the effects of 2-deoxyglucose inhibiting synovial pannus of adjuvant arthritis rats and to explore its potential mechanism of inhibiting angiogenesis by investigating proliferation, migration and matrigel tube formation assay .

METHODSThe effect of 2-DG on synovial pannus was evaluated by histopathology of HE staining; HUVEC proliferation was determined by CCK-8 method; migration of FLS were determined by transwell; matrigel tube formation assay was made for assessing tube number of HUVEC; p-AMPK and Bcl-2 were detected by Western blot assay; AMPK signaling pathway in HUVEC was inhibited by compound C, which is an inhibitor of AMPK activation.

RESULTS2-DG (200 mg/kg) obviously decreased appearance of synovial pannus ( < 0.01); , 2-DG (0.5 mmol/L and/or 5 mmol/L) obviously inhibited proliferation, migration and tube number of HUVEC ( < 0.01 or < 0.001), and its effects on HUVEC were reversed by using AMPK antagonist (Compound C); Western blot showed that 2-DG (5 mmol/L) increased expression of p-AMPK and decreased expression of Bcl-2 ( < 0.05).

CONCLUSIONSActivating AMPK pathway and decreasing expression of Bcl-2 may the potential mechanism by which 2-DG contributes to anti-angiogenesis and effects of inhibiting proliferation, migration and tube number of HUVEC.

Objective To study the protective effect of chlorogenic acid(CGA)on the apoptosis of PC12 cells induced byβ-amyloid protein23-35(Aβ25-35)and its mechanism. Methods The cells model of death was estab-lished by Aβ25-35 (20 μmol/L)-induced PC12 cells. The cells were interfered with 5 different concentrations of CGA. CCK-8 assay was used to detect cells viability to determine the 3 concentrations of CGA in future experi-ments. The cells were divided randomly into control group ,model group and interference groups with 3 different concentrations of CGA. Cells apoptosis rates were detected by flow cytometry;colorimetry method was used to detect MDA,SOD and GSH-Px. The mitochondrial membrane potential(MMP)was detected by fluorescent staining and the expression of caspase-3 by western blot. Results Compared with model group,the cells viability of CGA groups were increased but the apoptosis rates were reduced;the activity of SOD and GSH-Px were increased but the level of MDA,MMP and caspase-3 were decreased(P<0.05). Conclusions CGA has a protective effect on Aβ25-35-induced PC12 cells apoptosis and it may be related to the improvement of cellular antioxidation capacity and mitochondrial damage.

OBJECTIVE: To explore the effects of 3-methyladenine (3-MA, an autophagy inhibitor) on sensitivities of nasopharyngeal carcinoma cells to radiotherapy and chemotherapy and the underlying mechanisms.
 METHODS: Cell proliferation was examined by MTT and colony formation assay, while cell apoptosis was evaluated by annexin V/PI double staining and 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) staining. Mitochondrial membrane potential was measured by commercial kit (JC-1). The expression of endoplasmic reticulum stress (ERS)-related protein, glucose-regulated protein 78 (GRP78) and autophagy-related protein beclin1, microtubule-associated protein 1 light chain 3 (LC3) were examined by Western blot.
 RESULTS: Cisplatin (DDP), ionizing radiation (IR) or tunicamycin (TM) treatment obviously inhibited the proliferation of HONE-1 cells in a concentration-dependent and time-dependent manner. Compared with control group, pretreatment with 1 mmol/L of 3-MA significantly 
reduced cell viability and enhanced the apoptosis in the DDP (6.00 μmol/L), 4.00 Gy IR or TM (1.00 μmol/L) groups. There was no significant difference in the apoptosis between the DDP (5.8%) and 4Gy IR (6.7%) groups. Compared with the control group, protein levels of GRP78, beclin1 and lipid-conjugated membrane-bound form (LC3-II) were significantly increased after the treatment of DDP, 4.00 Gy IR or TM, which were inhibited by pretreatment of 3-MA.
 CONCLUSION 3-MA can sensitize HONE-1 cells to chemotherapy and radiotherapy, which is related to prevention of endoplasmic reticulum stress-induced autophagy in nasopharyngeal carcinoma cells.
Adenine ; analogs & derivatives ; pharmacology ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; Beclin-1 ; Carcinoma ; Cell Line, Tumor ; drug effects ; radiation effects ; Cell Proliferation ; Cell Survival ; Cisplatin ; pharmacology ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; metabolism ; Humans ; Membrane Potential, Mitochondrial ; Membrane Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; pathology ; Radiation, Ionizing ; Radiation-Sensitizing Agents ; pharmacology ; Tunicamycin ; pharmacology

OBJECTIVETo investigate the effect of chloroquine in reversing multidrug resistance (MDR) of HNE1/DDP cell line and explore the mechanism.

METHODSMTT assay was used to detect the cell viability of HNE1 and HNE1/DDP after exposure to different concentrations of DDP (2, 4, 8, 16 and 32 µmol/L) and different concentrations of chloroquine (5, 10, 20, 40, and 80 µmol/L). q-PCR was used to assess the expression of MDR1 mRNA and Western blotting was employed to detect P-glycoprotein (P-gp) expression in HNE1 and HNE1/DDP cells exposed to 5 and 10 µmol/L chloroquine. The cell apoptosis rate of HNE1 and HNE1/DDP cells exposed to 10 and 20 µmol

RESULTSChloroquine exposure caused dose-dependent suppression of the proliferation in both HNE1 and HNE1

CONCLUSIONChloroquine can reverse multidrug resistance in HNE1
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; Cell Line, Tumor ; Chloroquine ; pharmacology ; Down-Regulation ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans

Objective To investigate the effect of chloroquine in reversing multidrug resistance (MDR) of HNE1/DDP cell line and explore the mechanism. Methods MTT assay was used to detect the cell viability of HNE1 and HNE1/DDP after exposure to different concentrations of DDP (2, 4, 8, 16 and 32μmol/L) and different concentrations of chloroquine (5, 10, 20, 40, and 80μmol/L). q-PCR was used to assess the expression of MDR1 mRNA and Western blotting was employed to detect P-glycoprotein (P-gp) expression in HNE1 and HNE1/DDP cells exposed to 5 and 10μmol/L chloroquine. The cell apoptosis rate of HNE1 and HNE1/DDP cells exposed to 10 and 20μmol/L chloroquine was determined by PI assay. Results Chloroquine exposure caused dose-dependent suppression of the proliferation in both HNE1 and HNE1/DDP cells, and significantly reversed multidrug resistance in HNE1/DDP cells. The expressions of MDR1 mRNA and P-gp protein were significantly lowered in the cells treated with chloroquine. Conclusion Chloroquine can reverse multidrug resistance in HNE1/DDP cells possibly through down-regulation of MDR1 and inhibition of P-gp protein.

Objective To investigate the effect of chloroquine in reversing multidrug resistance (MDR) of HNE1/DDP cell line and explore the mechanism. Methods MTT assay was used to detect the cell viability of HNE1 and HNE1/DDP after exposure to different concentrations of DDP (2, 4, 8, 16 and 32μmol/L) and different concentrations of chloroquine (5, 10, 20, 40, and 80μmol/L). q-PCR was used to assess the expression of MDR1 mRNA and Western blotting was employed to detect P-glycoprotein (P-gp) expression in HNE1 and HNE1/DDP cells exposed to 5 and 10μmol/L chloroquine. The cell apoptosis rate of HNE1 and HNE1/DDP cells exposed to 10 and 20μmol/L chloroquine was determined by PI assay. Results Chloroquine exposure caused dose-dependent suppression of the proliferation in both HNE1 and HNE1/DDP cells, and significantly reversed multidrug resistance in HNE1/DDP cells. The expressions of MDR1 mRNA and P-gp protein were significantly lowered in the cells treated with chloroquine. Conclusion Chloroquine can reverse multidrug resistance in HNE1/DDP cells possibly through down-regulation of MDR1 and inhibition of P-gp protein.

Aim Toexploretheinhibitioneffectof triptolide on nasopharynx cancer, and the mechanism. Methods Theinhibitionofcellproliferationwasde-tected by MTT assay;the cell apoptosis was analyzed by flow cytometry with propidium iodide staining. The ex-pressions of glucose regulated protein 78 ( GRP-78 ) , Akt and pAkt in cells were examined by Western blot;the effect of triptolide on reactive oxygen species ( ROS) accumulation was detected by ROS Fluorescent Probe-DHE.Results MTTassayshowedthatthe growth of nasopharynx cancer was inhibited by triptol-ide , and the inhibition occurred in a dose and time-de-pendent manner following triptolide treatment in CNE-2Z nasopharynx cancer cells. Propidium iodide staining revealed that the apoptosis of CNE-2 Z cells was in-duced remarkably by triptolide. After CNE-2Z cells treated with 25, 50,100 nmol·L-1 of triptolide for 24 h, the apoptosis rate was 14%,26. 9% and 34. 4% re-spectively. Western blot experiment showed that the expression of GRP-78 had no significant change follow-ing triptolide treatment in CNE-2 Z nasopharynx cancer cells for 24 h, but the expression and the phosphoryla-tion level of Akt were strikingly decreased. The experi-ment of ROS uncovered that CNE-2 Z nasopharynx cancer cells increased generation of ROS after treat-ment with triptolide for 4 hours, and acted cells in a dosedependentmanner.Conclusions Triptolidecan inhibit the growth of CNE-2 Z nasopharynx cancer cells in a dose and time-dependent maner. The mechanism may be related with the point that triptolide can induce oxidative stress, incease ROS, inhibit the expression and the phosphorylation level of Akt,then promote the apoptosis of CNE-2Z cells.