Objective To evaluate effects of simultaneous inhibition of mammalian target of rapamycin complex 1(mTORC1)kinase and glycogen synthase kinase-3β(GSK-3β)on phosphorylation of 4E-binding protein-1(4EBP1), cap-dependent translation, as well as survival and apoptosis of melanoma cells. Methods Cultured A375 cells were classified into several groups to be treated with dimethyl sulfoxide (DMSO group), the mTORC1 kinase inhibitor everolimus at a concentration of 5 nmol/L (everolimus group), the GSK-3β kinase inhibitor AR-A014418 at a concentration of 10 μmol/L (AR-A014418 group), or 5 nmol/L everolimus and 10 μmol/L AR-A014418(combined treatment group). After additional culture, Western-blot analysis was performed to measure protein expressions of phosphorylated 4EBP1 (p4EBP1)and survivin in A375 cells, m7GTP pull down assay to estimate interaction between eukaryotic initiation factor-4E (eIF4E)and eIF4G, cell counting kit 8 (CCK8)assay to evaluate cell proliferation, and flow cytometry to detect cell apoptosis. Results Both everolimus and AR-A014418 had inhibitory effects on 4EBP1 phosphorylation and survivin expression. The expressions of p4EBP1-65 and survivin were both significantly decreased in the everolimus group (0.74 ± 0.05 and 0.71 ± 0.06 respectively), AR-A014418 group (0.62 ± 0.06 and 0.58 ± 0.07 respectively)and combined treatment group (0.14 ± 0.04 and 0.09 ± 0.05 respectively)compared with the DMSO group (1.00 ± 0.07 and 1.00 ± 0.06, respectively, all P < 0.001), with the most significant decrease observed in the combined treatment group. As m7GTP pull-down assay showed, the everolimus group, AR-A014418 group and combined treatment group all showed significantly lower relative expression levels of eIF4G(0.72 ± 0.04, 0.67 ± 0.05 and 0.12 ± 0.05 vs. 1.00 ± 0.06, all P < 0.001), but significantly higher relative expression levels of 4EBP1 (1.98 ± 0.16, 2.32 ± 0.17 and 7.58 ± 0.25 vs. 1.00 ± 0.08, all P < 0.001)than the DMSO group, and the combined treatment group showed the lowest eIF4G expression but highest 4EBP1 expression. After 24-hour culture, the proliferation of A375 cells was inhibited by 18.5% ± 1.3% in the everolimus group, 19.8% ± 1.8% in the AR-A014418 group, and 61.2% ± 2.1% in the combined treatment group compared with the DMSO group, with the strongest inhibition noted in the combined treatment group. The inhibitory effects of everolimus and AR-A014418 on cell proliferation increased over time, and showed the same trend at 48 hours. Flow cytometry showed that the apoptosis of A375 cells was accelerated by the 24-hour treatments with everolimus and AR-A014418 alone or in combination, with the apoptosis rate being 14.28% ± 2.18%, 14.57% ± 2.35% and 55.18% ± 6.27% in the everolimus group, AR-A014418 group and combined treatment group respectively, and the combined treatment showed the strongest accelerating effect. Conclusion The combined treatment with everolimus and AR-A014418 can evidently inhibit 4EBP1 phosphorylation and eIF4F complex formation in A375 cells, which then suppress cap-dependent translation and promote apoptosis of melanoma cells.

Body Mass Index ; Humans ; Male ; Middle Aged ; Obesity, Abdominal

Objective To evaluate the risk factors of septic shock after mini-percutaneous nephrolithotripsy (mPCNL). Methods Clinical data of 1 590 cases who underwent mPCNL from January 2013 to December 2014 were retrospectively analyzed. The x2 test and logistic regression were used to identify the key risk factors for septic shock after mPCNL. Results Of the 1 590 patients, 18 patients suffered septic shock, including 6 male patients and 12 fe﹣male patients. Their mean age was (45.6 ± 13.5) years (28 ~ 69 years). White cell in urine was 100 percent, the stone diameter ranged from 1.5 to 5.0 cm, unichannel for 15 cases while multichannel for 3 cases, the operation du﹣ration ranged from 45 to 200 min, mean (87.0 ± 56.0) min. 2 in 18 cases died in multiple organ failure, the others recovered till discharged. In x2 test, female gender (P = 0.001), (+++ ~ ++++) white cells in urine (P= 0.042), un-preoperative nephrostomy drainage (P=0.041) had significant association with septic shock after mPCNL. While in multivariate analysis, female gender ( O? = 5.471, 95 % CI: 0.756~21.452, P< 0.05) and un-preoperative nephrostomy drainage (O? =3.106, 95%CI:1.283~7.907, P<0.05) were identified as independent risk factors for septic shock after mPCNL. Conclusions Female gender and un-preoperative nephrostomy drainage are the key risk factors for septic shock after mPCNL.

ObjectiveTo evaluate the methods in treatment of upper urinary calculi in a low-head lithotomy with right or left lateral tilt by an ureteroscopy approach. Methods From September 2009 to May 2015, 110 patients with upper ureteral calculi (after failed ESWL) were underwent holmium laser lithotripsy by a ureteroscopy approach in a low-head lithotomy with right or left lateral tilt. Their clinical data and complications were analyzed retrospectively. Results Surgical effect of patients was satisfied with the success of gravel 91 patients, with a total rate of 82.7 %gravel. Conclusion Low-head lithotomy with right or left lateral tilt is a good body position to perform ureteroscopic lithotripsy for upper ureteral calculi. It is safe and effective.

Objective To investigate the effect of Zingiber corallinum oil ( ZCO ) on apoptosis and proliferation of cervical carcinoma cell line HeLa. Methods HeLa cells were treated with different concentrations of ZCO(5-80 mg·L-1)in vitro. Cytotoxicity rate was determined by CCK-8 assay. The morphological changes was observed using inverted microscope after AO/EB staining. Caspase-3 activities were measured with a colorimetric method. Protein level of hsp-70 were detected by Western blotting. Cell cycle and apoptosis were analyzed by flow cytometer ( FCM ) . Results ZCO exhibited effect of proliferation inhibition and apoptosis-inducing on the growth of HeLa cells in a dose-dependent manner. Caspase-3 activities increased in a dose-dependent manner while the expression of hsp-70 decreased. Cell cycle was arrested in G2/M phase. Conclusion ZCO exhibites a marked effect of proliferation inhibition and apoptosis-inducing on HeLa cells. The mechanism of ZCO might be activating the key enzyme in apoptotic pathway, so that the expression of hsp-70 is down-regulated, and cell cycle is arrested in G2/M phase.

Objective To evaluate the effect of neoadjuvant chemotherapy (NAC) by investigating the expression of cyclin D1 in human breast cancer before and after NAC.Methods Eighty-four cases of breast cancer were diagnosed by core biopsies.The expression level of cyclin D1 in cancer tissues was measured by immunohistochemical envision two-step method before and after NAC (pirarubicin and docetaxel regimen for 3-4 cycles).Results Complete remission (CR) occurred in 4 cases of 84 patients (4.76 ％) with 2 pathological complete response cases,partial response (PR) in 54 cases (64.29 ％),stability (SD) in 26 cases (30.95 ％) and no disease progression (PD) patients.The positive rate of cyclin D1 in cancer tissues [65.48 ％ (55/84)] was significantly decreased after NAC [39.29 ％ (33/84)] (x2 =11.55,P =0.001).In clinical level,the ease rate was significantly improved in patients whose cyclin D1 expression switched from positive [86.36 ％ (19/22)] to negative [45.45 ％ (15/33)] after NAC treatment (x2 =9.359,P =0.002).Conclusion NAC significantly decreases the expression of cyclin D1 in breast cancer tissues.Meanwhile,the ease rate is improved when cyclin D1 expression switched from positive to negative after NAC.Therefore,cyclin D1 expression can be used as an evaluation index for the efficiency of NAC.

Objective To investigate the relationship between waist circumference and new-onset non-alcoholic fatty liver disease in non-obese patients with diabetes mellitus. Methods A total of 1 950 patients with diabetes mellitus, who determined fasting plasma glucose(FPG)≥7.0 mmol/L or who were using hypoglycemic drugs and FPG<7.0 mmol/L,and body mass index (BMI)< 25 kg/m2, was selected in this study using prospective cohort method. Patients were divided into five groups according to the baseline data of waist circumference, including waist circumference<78 cm (A group, n=387), 78 cm90 cm (E group, n=421). Multiple Logistic regression model was used to analyze influential factors of new-onset non-alcoholic fatty liver disease in non-obese patients with diabetes mellitus. Re?sults The average duration of follow-up was(47.24±5.13) months. The incidence rate was 11.85%(231/1 950) in patients with non-alcoholic fatty liver disease. The incidence rates were 6.98%, 9.28%, 12.38%, 14.19%and 15.68%in A, B, C, D and E groups, and which were increased with the increased waist circumference (P<0.05). Results of multiple Logistic re?gression model analysis showed that compared with A group,OR values were 1.97 and 2.19 in D and E groups respectively (P<0.05). Conclusion Waist circumference≥85 cm was the risk factors of new-onset non-alcoholic fatty liver disease in non-obese patients with diabetes mellitus.

Objective To study the awareness of perimenopausal hormone replacement therapy (HRT) among a part of the medical care personnel in Guiyang . Methods A survey was conducted among 500 medical staff members in 4 hospitals of Guiyang by cluster random sampling using questionnaire about HRT. Results The survey showed that 74.6% (373/500) medical staff thought that the hormone replacement therapy was necessary to perimenopausal women; 96.7% (87/90) of obstetrics and gynecology doctors believed that it was necessary for perimenopausal women to use HRT,which was significantly higher than the doctors of other specialties 68.6% (166/242) and the nurses group 71.4% (120/168) (χ2=28.509, 23.537, P<0.01). Only 5.8%(29/500) of the medical personnel were willing to recommend HRT. In light of the attitude for recommending HRT, the obstetricians and gynecologists group was more significantly higher than the other specialties doctors group (χ2=86.781, P<0.01). Conclusion The knowledge of hormone replacement therapy in part of Guiyang medical personnel is not sufficient;the recommending rate of HRT was low;the side effects of HRT was still a concern. There are differences between obstetrics and gynecology doctors and doctors other specialties and nurses in HRT knowledge.

With a global rise in morbidity rates， obesity has become a pressing public health issue. With increased adipocyte number and volume as the main characteristics， obesity is also manifested by metabolic disorders to varying degrees. At the same time， obesity is a risk factor for diabetes， hypertension， stroke， cancer， and cardiovascular diseases， imposing burdens on society and families. Influenced by lifestyle， environment， behavior， and genetics， obesity is caused by the interaction of many factors， and its pathological process is complex， involving inflammation， autophagy， and intestinal dysbiosis. The mitogen-activated protein kinase （MAPK） cascade reaction， a pivotal signaling pathway， plays a crucial role in cellular processes such as proliferation， differentiation， apoptosis， and stress responses. Both Chinese and international studies indicate that the MAPK signaling pathway can effectively regulate obesity through various pathways， including the modulation of adipocyte differentiation and apoptosis， appetite control， and inflammation improvement. Moreover， traditional Chinese medicine （TCM） has demonstrated significant efficacy in preventing and treating obesity， leveraging advantages such as multiple targets， diverse components， and minimal adverse effects. Research indicates that the MAPK signaling pathway is a primary focus of TCM regulation in this context， although a systematic review in this field is currently lacking. Therefore， this paper， by reviewing the latest Chinese and international research， provided a concise overview of the basic structure of the MAPK pathway， with a specific emphasis on recent progress in TCM interventions targeting the MAPK pathway for obesity treatment. The results indicate that regulating adipose tissue formation， differentiation， and thermogenesis， reducing inflammation and oxidative stress levels， and improving insulin sensitivity and metabolic disorders seem to be the main ways for TCM to regulate the MAPK pathway to prevent and treat obesity. However， it is necessary to find more research methods and explore potential mechanisms underlying TCM formulations based on the MAPK pathway for obesity prevention and treatment.

RNF20, an E3 ligase critical for monoubiquitination of histone H2B at lysine 120 (H2Bub), has been implicated in the regulation of various cellar processes; however, its physiological roles in adipocytes remain poorly characterized. Here, we report that the adipocyte-specific knockout of Rnf20 (ASKO) in mice led to progressive fat loss, organomegaly and hyperinsulinemia. Despite signs of hyperinsulinemia, normal insulin sensitivity and improved glucose tolerance were observed in the young and aged CD-fed ASKO mice. In addition, high-fat diet-fed ASKO mice developed severe liver steatosis. Moreover, we observed that the ASKO mice were extremely sensitive to a cold environment due to decreased expression levels of brown adipose tissue (BAT) selective genes, including uncoupling protein 1 (Ucp1), and impaired mitochondrial functions. Significantly decreased levels of peroxisome proliferator-activated receptor gamma (Pparγ) were observed in the gonadal white adipose tissues (gWAT) from the ASKO mice, suggesting that Rnf20 regulates adipogenesis, at least in part, through Pparγ. Rosiglitazone-treated ASKO mice exhibited increased fat mass compared to that of the non-treated ASKO mice. Collectively, our results illustrate the critical role of RNF20 in control of white and brown adipose tissue development and physiological function.