[Summary] Considerable efforts have been made to understand the cellular and molecular mechanisms of metformin ,a potent antihyperglycemic agent now recommended as the first‐line treatment of patients with T2DM. The main effect of this drug is to reducethe hepatic glucose output ,primarily through inhibition of the mitochondrial respiratory chain complex Ⅰ and then activating AM P‐activated protein kinase (AMPK) ,which provide a generally acceptable mechanism for the action of metformin on hepatic gluconeogenesis. Beyond its effect on glucose metabolism ,metformin has been reported to improve ovarian function of patients with polycystic ovary syndrome (PCOS) and to reduce the risk of microvascular and macrovascular complications in patients with T2DM. In addition ,metfomin has also recently been suggested as an adjuvant treatment of cancer. Here we reviewed the progress of mechanism research and clinical application of metformin.

Background Accurate measurement of corneal refractive parameters plays an important role in the diagnosis of eye diseases,design of refractive surgery and calculation of intraocular lens (IOL) power.Ophtha TOP is a new IOL calculator,so it is necessary to evaluate the repeatability and accuracy of Ophtha TOP in measuring corneal refractive parameters.Objective This study was to evaluate the repeatability of Ophtha TOP in measuring corneal refractive status in normal eyes and determine the agreement of the measuring outcomes between Ophtha TOP and Pentacam in pre-surgery cataract eyes.Methods A reliability evaluation method of diagnosis test was performed from September 2013 to October 2014.Sixty eyes of healthy volunteers aged (30.83 ±8.66) years old were examined in Tianjin Eye Hospital with Ophtha TOP for more than 5 times to evaluate the stability in measuring corneal refractive parameters.Corneal refractive parameters were measured by both Ophtha TOP and Pentacam for more than twice in 30 pre-surgery cataract eyes to assess the agreement with Pentacam.Keratometry and astigmatism parameters were recorded,including the flattest meridian (Kf),steepest meridian (Ks),mean keratometry (Km) and the astigmatism at J0 and J45.Intra-class correlation coefficients (ICC),repeatability (2.77Sw) and coefficient of variation (CoVs) were used to assess the stability of Ophtha TOP,and Bland-Altman graphs were adopted to value the consistency of measuring outcomes between Ophtha TOP and Pentacam.This study complied with Helsinki declaration,and written informed consent was obtained from each patient prior to any medical examination.Results The ICC of Ks,Kf,Km,J0 and J45 by Ophtha TOP were all ≥0.90,and 2.77Sw were all ≤0.36,The CoVs of Kf,Ks,Km by Ophtha TOP were all ≤0.30.The measuring values of Kf,Ks and Km were significantly higher in the Ophtha TOP than those in the Pentacam,with the mean differences (0.18±0.28) D,(0.24±0.29) D and (0.21± 0.26) D,respectively(t =3.48,4.50,4.49,all at P=0.00).The mean difference of the J0 was (0.07±0.21) D and that of the J45 was (-0.02±0.18)D between Ophtha TOP and Pentacam,showing insignificant differences between them (both at P＞0.05).Bland-Altman graphs revealed that the 95％ limit of agreement (LoA) of Kf,Ks and Km between the two methods was-0.37 D to 0.73 D,-0.33 D to 0.81 D and-0.30 D to 0.72 D,respectively,and that of J0 and J45 was-0.34 D to 0.48 D and-0.37 D to 0.33 D,respectively.Conclusions Ophtha TOP shows an excellent repeatability.Compared with Pentacam,Ophtha TOP presents a good consistence for measurement of the corneal astigmatism and moderate consistence for corneal curvature.

恶性黑色素瘤的发病率和病死率在逐年增长，引发广泛关注。目前恶性黑色素瘤的治疗方式主要为手术、化疗和靶 向治疗，总体疗效极差。然而，PD-1抗体治疗恶性黑色素瘤的成功，使广大研究者把希望聚焦于生物免疫治疗。肿瘤浸润淋巴细 胞（tumor infiltrating lymphocyte，TIL）是从肿瘤组织分离出的淋巴细胞，经体外扩增后得到的能够杀伤肿瘤细胞的细胞群，具有 高杀瘤活性和高靶向性特点。临床试验研究发现，TIL对恶性黑色素瘤治疗效果显著且稳定，但其临床疗效尚未完全阐明。本文 对TIL免疫治疗的发展及其在恶性黑色素瘤治疗中的研究新进展作一综述，以期为恶性黑色素瘤的治疗提供新的策略。

Objective To investigate the effects of curcumin on mRNA expression of cytokines related to Toll-like receptor (TLR) 4 signaling in THP-1 cells.Methods After pretreatment with different concentrations (50,25,12.5 mg/L) of curcumin or dexamethasone for 12 hours,THP-1 cells were stimulated by lipopolysaccharide (LPS.1 mg/L) for 4 hours followed by the collection of cells.Then total RNA was isolated from these cells and subjected to reverse transcription-polymerase chain reaction (RT-PCR) for the detection of mRNA expression of tumor necrosis factor receptor-associated factor 6 (TRAF6),interleukin-1 receptorassociated kinase (IRAK1) and nuclear factor (NF)-κB.THP-1 cells without pretreatment or stimulation served as negative control,and those only stimulated with LPS served as LPS group.Results After stimulation with LPS (1 mg/L) for 4 hours,the mRNA expressions of TRAF6,IRAK1 and NF-κB were significantly upregulated in THP-1 cells compared with negative control cells (f=38.69,39.13,23.99,all P<0.01).Curcumin of 50 mg/L and 25 mg/L significantly inhibited the mRNA expressions of TRAF6.IRAK 1 and NF-κB upregulated by LPS with an inhibition rate of more than 50% (all P<0.0 1).Conclusions Certain concentrations of curcumin can inhibit the mRNA expressions of TRAF6.IRAK1 and NF-κB.which demonstrates the regulatory effect of curcumin on the mRNA expressions of TLR4 signaling pathway-associated cytokines.

Objective To investigate the molecular mechanism of vascular endothelial growth factor ( VEGF) expression in bronchial epithelial cells (Beas-2B)induced by particulate matter 2.5(PM2.5).Methods PM2.5 powder was dis-solved in DMEM medium and diluted into five concentrations , 0,12.5,25,50 and 100μg/ml, respectively.The double an-tibiotics ( streptomycin and penicillin ) and FBS were added into the solution to a 2% final concentration of serum system after being treated by ultrasound for 30 minutes.The cultured Beas-2B cells were then treated with different doses of PM2.5.Subsequently, nuclear factor-kappa B(NF-κB) transactivity and the transcriptional activation of vegf gene promot-er were tested by dual-luciferase reporter gene analysis system while phosphorylation of p 65 , expression levels of IκBαand VEGF were detected by Western blotting .Results PM2.5 induced up-regulation of VEGF expression in Beas-2B cells in a dose-dependent manner , accompanied by NF-κB transactivation at the highest level under 100 μg/ml of PM2.5 treatment. Moreover, PM2.5 induced degradation of the repressor protein IκBαand increase in the phosphorylation level of p 65 sub-unit in Beas-2B cells.Knockdown of NF-κB p65 expression significantly inhibited vegf gene promoter transcriptional activa-tion as well as VEGF protein expression in Beas-2B cells induced by PM2.5.Conclusion PM2.5 induces VEGF expres-sion via activation of NF-κB pathway in bronchial epithelial cells .

Objective To investigate mRNA expressions of apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 family (APOBEC3s or A3s) as well as expressions and subcellular distribution of major A3 proteins in HaCaT keratinocytes carrying the genome of human papillomavirus type 11 (HPV11.HaCaT),and to evaluate regulatory effects of exogenous interferon-alpha (IFN-α) on the expressions of A3s.Methods The basal levels of A3A,A3B,A3C and A3H mRNA expressions were measured by real-time fluorescence-based quantitative PCR (qRT-PCR) in HPV11.HaCaT cells and normal HaCaT cells.Cultured HaCaT,HPV11.HaCaT and Hela cells were treated with recombinant human IFN-α 2b (rhIFN-α2b) at concentrations of 104,105 and 106 IU/ml for 6,24 and 48 hours separately,and those receiving no treatment served as the normal control groups.Then,qRT-PCR was performed to measure mRNA expressions of A3A,A3B,A3C and A3H in these cells.Immunofluorescence staining was conducted to observe the expression and distribution of A3A protein in cells after the treatment with rhIFN-α2b for 6 hours.Results As qRT-PCR showed,the basal levels of A3A,A3B and A3C mRNA expressions were all significantly higher in HPV11.HaCaT cells than in normal HaCaT cells (all P ＜ 0.05).After stimulation,the mRNA expressions of the four A3 members increased to different extents with the increase in rhIFN-α2b concentrations,and the increase in A3A mRNA was the most significant.Compared with corresponding normal control groups,the mRNA expression of A3A was significantly increased in HaCaT cells (35.77 ± 5.01 vs.1.00 ± 0.05,P ＜ 0.05),HPV 11.HaCaT cells (15.34 ± 2.14 vs.0.99 ± 0.01,P ＜ 0.05) and Hela cells (24.60 ± 5.45 vs.0.97 ± 0.03,P ＜ 0.05) after the treatment with rhIFN-α2b at 106 IU/ml for 6 hours,while the increase in A3B,A3C and A3H mRNA expressions was no more than 9-fold in these cell lines after that.Enhanced staining for A3A was observed in nuclei and cytoplasm of the 3 cell lines after the treatment with rhIFN-α2b at 106 IU/ml for 6 hours.Conclusions HPV11 transfected into HaCaT cells can activate intracellular A3s,especially A3A.IFN-α may play an immunoregulatory role by inducing high levels of A3A expression.

Objective To quantify the changes of tongue color with the development of the disease according to the characteristics of human vision.Methods Psychological scales of tongue color perception were obtained from thc psychophysical experiments.Polynomial regression and support vector regression (SVR) were used to establish the mathematical model between the physical values and the psychological scales of tongue color.Results The psychological scales exported from the model could correspond to the visual perception of tongue color change, and SVR has higher accuracy.Conclusions The psychological scale, can not only quantitatively evaluate the tongue color in the development of the disease, but also quantify the degree of disease, to assist the doctors for disease diagnosis and treatment.

Objective To investigate therapeutic effect of the low dose clomiphene treatment of idiopathic oligoasthenospermia and effect on sperm epididymal protein level 4.Methods183 cases of idiopathic oligoasthenospermia were divided into three groups, vitamin group of 61 patients were given vitamin E,clomiphene group(61 cases)treated with low dose clomiphene, combined group of 61 patients were given low dose clomiphene and vitamin E, lasted for 12 weeks.Semen volume, sperm density, sperm motility, sperm survival rate and sperm reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), human epididymis protein 4 (HE4), follicle stimulating hormone, luteinizing hormone, testosterone concentration were observed before and after treatment.After treatment, the patients were followed up for 1 year, the observation of the natural pregnancy rate after treatment.ResultsSperm density, sperm survival rate and sperm and a A+B grade sperm in the clomiphene group were significantly higher comparedwith the vitamin group (P< 0.05), sperm density, sperm survival rate and sperm and a A+B grade sperm in the combined group were significantly increasedcompared with clomiphene group and vitamin group (P< 0.05),levels of HE4 after treatment was significantly lower compared with before treatment (P< 0.05), And the difference between the three groups were statistically significant (P< 0.05).Follicle stimulating hormone, luteinizing hormone, testosterone in clomiphene group and combined group after treatment, were significantly increased compared with before treatment (P< 0.05), and the combined group had no statistically difference compared with clomiphene group.The pregnancy rate in 1 year of vitamin group, clomiphene group and combined group were 24.59%, 42.62% and 62.29% (P< 0.05).ConclusionClomiphene treatment of idiopathic asthenospermia is better than vitamin E, the combination of the two has a synergistic effect.

Objective To explore the association of COX-2 Gly587Arg polymorphism with the risk of primary liver cancer.Methods Two hundred and seventy patients with primary liver cancer and 540 health people were selected as our subjects.DNA were extracted from peripheral blood lymphocytes,and genotypes were measured by polymerase chain reaction-restriction fragment length polymorphism method.Odds ratios(OR) and 95％ confidence intervals(CI) were estimated by logistic regression.Results Two kinds of genotype (587Gly/ Gly and Gly/Arg) were found in all participants.No one carried 587Arg/Arg genotype.Among primary liver cancer patients,91.5％ (247/270,) 8.5％ (23/270) of individuals carried 587Gly/Arg and Gly/Arg genotype,which was significantly higher than that of controls (96.5％ (521/540,) 3.5％ (19/540)).Multivariate Logistic regression analysis showed that individual carried 587Gly/Arg genotype had an increased risk of developing primary liver cancer (OR =2.56,95％ CI =1.37-4.79,P =0.003) compared with 587Gly/Gly carriers.Conclusion COX-2 Gly587Arg polymorphism is a risk factor for primary liver cancer in Han.

Objective To investigate the effects of 13-hexyl-berbefine hydroehlofide (HB-13) and 13-hexyl-paimatine hydrochloride (HP-13) on the activation of nuclear factor-kappa B (NF-kB) and phosphorylation of p38 mitogen-activated protein kinase (MAPK) in a human keratinocyte cell line, HaCaT stimulated by tumor necrosis factor alpha (TNF-α). Methods HaCaT cells were cultured in the presence of various concentrations (0.39, 0.78, 1.56 μg/mL) of HB-13 or HP-13 for 120 minutes followed by the stimulation with recombinant human TNF-α for 120 minutes (in phosphorylatEd-IkB-α test) or 15 minutes (in phosphorylated-p38 test). Then, HaCaT cells were disrupted, total protein was extracted, and the expressions of phosphorylated I B-α and phosphorylated p38 were detected with Western blot. HaCaT cells receiving neither pretreatment nor stimulation served as blank control, untreated HaCaT cells stimulated by rhTNF-α as stimulator control, and HaCaT cells pretreated with turmeric root tuber and stimulated by rhTNF-α as positive control. Results From 0.39 to 1.56 μg/mL, both HB-13 and HP-13 significantly inhibited the expression of p-IkB-α in HaCaT cells stimulated by rhTNF-α, and a nonsignificant dose-dependent trend was observed for their inhibitory effect, with the ICo value being 0.441 μg/mL for I-IB-13 (r = -0.990, n = 3, P > 0.05) and 0.832 μg/mL for HP-13 (r = -0.992, n = 3, P > 0.05). In contrast, neither 1-113-13 nor HP-13 within the experiment concentration range had a significant effect on the expression of p-p38 in HaCaT cells stimulated by rhTNF-α (P > 0.05). Conclusions Within the experimental concentration range, both HB-13 and HP-13 can inhibit the activation of NF-kB in HaCaT cells induced by TNF-α signal, but neither of them suppress the phosphorylation of p38MAPK induced by TNF-α signal in HaCaT cells.