Background Recombinant hirudin variant Ⅲ(rHV3) can effectively prevent galactose-induced human lens epithelial cells LECs injury,but little is known about the molecular mechanism of its action.Objective The present study was to investigate the effects of rHV3 on the expression of apoptosis-related genes in damaged LECs induced by galactose.Methods The rHV3 was extracted by our research group,and the biological activity of rHV3 was identified by titration of thrombase according to Markwardt's method.Human LECs (SRA01/04) were cultured using 125×10-3 mol/L D-galactose+10% FBS+D/F12 medium to establish the damaged human LECs model.rHV3 was added into the medium of the damaged human LECs model.Human LECs were cultured in D/F12 medium containing 10% FBS as normal control.The expression of apoptosis-related genes,such as aldose reductase (AR),bax,bcl2 and p53,in LECs at the mRNA level was detected using RT-PCR.The abundance ratio of target genes was presented with the absorbance (A) of gene mRNA/GAPDH mRNA.Results Compared to the normal control group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were significantly elevated in model group (t=3.90E-06,t=8.44E-04,t=5.15E-08,P＜0.01).However,in the rHV3-treated group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were lower than those of model group (t=5.90E-06,t=1.51E-04,t=3.42E-06,P＜0.01).The bcl2 mRNA/GAPDH mRNA was markedly downregulated in the model group when compared with the normal control group (t=1.86E-05,P＜0.01);while after rHV3 addition,bcl2 mRNA/GAPDH mRNA increased in comparison with the model group (t=8.56E-05,P＜0.01).Conclusion 125×10-3mol/L D-galactose induces the damage and apoptosis of human LECs.rHV3 likely plays a protective function on D-galactose-induced damage of human LECs by inhibiting the polyol pathway and mitochondria-mediated pathway.

Female ; Humans ; Hyperacusis ; surgery ; Labyrinth Diseases ; surgery ; Middle Aged ; Otologic Surgical Procedures ; methods ; Retrospective Studies ; Semicircular Canals ; pathology ; surgery

Female ; Humans ; Infant ; Mitochondrial Proton-Translocating ATPases ; deficiency ; genetics ; Mutation

Objective To investigate the formation and special cell biological behavior of osteoclasts.Methods The live-cell imaging technology was adopted to consecutively and dynamically observe the whole process of multinuclear osteoclast formation induced by RANKL and M-CSF from rat peripheral blood monocyte.Meanwhile,the inverted phase contrast microscopy,TRAP staining,and scanning electron microscopy were also applied to identify the osteoclast.Results After 2-week cultivation,a great number of apocytes were found by the inverted phase contrast microscopy,and most apocytes and monocytes had positive reaction after TRAP staining.Moreover,many bone resorption lacunae in which osteoclasts were perhrming bone resorption function could be found in the bone slice under the scanning electron microscope.Live-cell imaging observation showed that the multinuclear osteoclasts were generated through self-fusion of monocytes,fusion of monocytes and apocytes,as well as fusion between apocytes.All fusion processes occurred under the condition of cell adherence.Time-lapse Microcinematography observation showed diverse shapes of osteoclasts and the cell division of multinuclear osteoclasts.Conclusion Rat peripheral blood monocyte can develop into osteoclast under induction of RANKL and M-CSF.Osteoclast can form gigantic apocyte via various types of cell fusion to increase its nucleus number and cell volume,vary its shape,and increase the area of plasma membrane.On the other hand,osteoclast can decrease its cell volume and nucleus number via cell division to adapt the needs of local morphology,biomechanics and bone resorption dynamics.It suggests that this non-mitosis cell division is a special cell biological behavior of osteoclast,which may be the basis of exerting its function and improving bone resorption efficiency.

ObjectiveTo optimize the condition of multiple fluorescence quantitative PCR and establish a new assay of four human herpes virus (HHV) detected by AllGlo quadruple fluorescence quantitative PCR.MethodsFour HHV including HSV-1,HSV-2,EBV and CMV were identified by sequence analysing the qualitative PCR production.Furthermore,they were quantitatively detected by AllGlo and TaqMan multiple fluorescence quantitative PCR respectively.ResultsBoth the positive rate and specificity of AllGlo and TaqMan in detecting single HHV achieved 100％.And AllGlo single fluorescence quantitative PCR prevailed over TaqMan's by Ct of 1-3.Four HHV can be simultaneously detected by AllGlo quadruple fluorescence quantitative PCR,comparing to the only two by TaqMan.ConclusionAllGlo fluorescence quantitative PCR assay allows a higher throughput,sensitivity and specificity than TaqMan in detection and thus provides a board prospect.

Objective To study the effects of p27kip1 gene on the apoptosis in esophageal carcinoma cell lines EC 9706. Methods A replication deficient adenovirus vector encoding p27kip1(ad p27kip1) was constructed and human esophageal carcinoma cell lines EC 9706 were transduced in vitro. The cell apoptosis were determined by flow cytometry, TUNEL technique and DNA fragmentation analysis. Results Ad p27kip1 containing the p27kip1 sequence was successfully constructed and the virus titer was 1.24?10 12 CFU/ml. The transduction efficiency was 100% when multplicity of infection≥50. FCM analysis revealed a sub G 1 cell peak in ad p27kip1 transduced esophageal carcinoma cell lines. Agarose electrophoresis showed marked ladder. The difference of apoptotic index between the ad p27kip1 group and the control group was statistically significant(37.3?3.4 vs. 1.3?0.2, P

Objective To investigate the effects of inhibiting ubiquitin proteasome pathway(UPP) on proliferation of gastric carcinoma cells and the possible mechanism was discussed. Methods The gastric carcinoma cell strain SGC 7901 was treated with MG 132 to inhibit its UPP specially. The effect of growth suppression on cells was evaluated with MTT assay. Cell cycle and apoptosis were detected by flow cytometry(FCM). DNA fragment analysis was used for confirming the presence of apoptosis. The activity of telomerase was examined by TRAP PCR ELISA. Expression of p27kip1 was detected by immunocytochemical technique. Results MG 132 had great inhibitory effect on the growth of SGC 7901 cells. The FCM analysis showed that the ratio of G0/G1 phase of control group was (46.3?4.1)%, the ratio of G0/G1 phase of SGC 7901 cells treated with MG 132 increased to (72.1?5.0)% ( P

Evidence has indicated that low doses of anti-tumor regimens can induce cell apoptosis in vitro, although different regimens induce apoptosis by different mechanism and pathway. In recent years, new tumor treatment strategy has been mainly focused on inducing tumor cell apoptosis. The present review discusses the advantages and disadvantages of inducing tumor cell apoptosis. The benefit of inducing apoptosis is not to cause inflammatory reaction, but as its disadvantage, it inhibits immune responses, and the phagocytosis of apopotic bodies may result in horizontal transfer of genes (including oncogenes and other oncogenic materials), which can be one of the causes of tumor relapse. This paper proposes that the tumor treatment strategy should be turn into promoting tumor cell necrosis and inducing anti-tumor immune responses.
Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Humans ; Necrosis ; chemically induced ; Neoplasms ; drug therapy ; immunology ; pathology

Adenocarcinoma ; metabolism ; pathology ; surgery ; Aged ; Female ; Gastrectomy ; methods ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; surgery ; Humans ; Keratins ; metabolism ; Neoplasms, Multiple Primary ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-kit ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; surgery

Diagnosis, Differential ; Female ; Humans ; Middle Aged ; Osteogenesis Imperfecta ; diagnosis ; therapy