BACKGROUND:Taurine is an important non-enzymatic system antioxidant in the lens.The mechanism of its anti-oxidative effect is mainly to protect lens from oxidative injury by anti-lipid peroxidation.OBJECTIVE:This study was to observe the effect of exogenous taurine on lens epithefial cell apoptosis-induced by H2O2 in vitro.DESIGN:A randomized controlled animal experiment.SETTING:Department of Ophthalmology,Affiliated Hospital of Qingdao University Medical College.MATERIALS:Seventy-five adult New Zealand standard tabbits,of either gender,weighing 1.5-2.5 kg,were provided by Qingdao Laboratory Animal Center.Reagent kit for in situ detecting cell apoptosis(Sigma Company,USA),taurine and H2O2(Shanghai Guangda Chemical Reagent Factory,China)were included in this study.METHODS:This study was performed in the Department of Ophthalmology and Central Laboratory.Affiliated Hospital of Qingdao University Medical College from Mav 2005 to June 2007.Rabbit lenses were harvested and randomly divided into 3 groups:control group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium which was renewed every 24 hours,H2O2 group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium containing 1 mmol/L H2O2 with addition of 62 μL H202(30 g/L)every 6 hours,and H2O2+taurine group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium containing 1 mmol/L H2O2 and 10 g/L taurine.which was renewed every 6 hours.The protocol was conducted in accordance with ethical guidelines for the use and care of animals.MAIN OUTCOME MEASURES:Observation of lens opacity 6.12,24.48 and 72 hours after culture;Lens epithelial cell apoptosis determined by DNA in situ end labeling and DNA fragment analysis.RESULTS:Lens opacity:The lens opacity in the H2O2 group was aggregrated gradually along with the time of oxidative injury.The lens opacity in the H2O2 group was severer than that in the H2O2+taurine group.Lens epithelial cell apoptosis:There were no apoptotic cells in the control group within 72 hours.The number of apoptotic cells in the H2O2 group was increased gradually with the prolonged time of oxidative injury.Till the 72nd hour,the cells were all tamed into apoptotic cells.A few aopototic celIs were found in the H2O2+taurine group since hour 24,and then were more and more,and the number of apoptotic cells accounted for about 30%at hour 72.The apoptotic rate in the H202+taurine group was significantly lower than that in the H2O group at each time point(q=8.6845,P＜0.01).There was no significant difference in apoptotic rate between the H202+taurine group and the control group (P＞0.05). Findings of DNA fragmentation assay:The DNA"ladder"was found in the H2O2 group at hours 24,36,48 and 72,while no DNA"ladder"but only normal electrophoresis straps were present in the other two groups 24 hours after culture.CONCLUSION:Taurine can inhibit the oxidative injury-induced apoptosis of rabbit lens epithelial cells,and alleviate lens opacity.

Objective To assess renal perfusion of chronic renal failure(CRF) at different stages in rabbits by contrast-enhanced ultrasound. Methods Fifteen rabbits were injected cationic bovine serum albumin(C-BSA) intravenously for 8 weeks to establish CRF models. Serum creatinine(Scr) and blood urea nitrogen(BUN) were determined before injection and 2,4,6,8 weeks after C-BSA injection respectively. The size of kidney was measured by two-dimensional ultrasound and contrast-enhanced ultrasound was performed on bilateral kidneys at the same time points. Renal perfusion was analyzed quantitatively with the time intensity curve. Results Renal cortical perfusion reduced started at 4weeks after injection,manifested as the peak signal intensity(PSI) of the time-intensity curve parameters decreased(P<0.05 or P <0.01). The speeds of perfusion and clearance of kidney were slower,showed as the time to peak intensity(PIT) and the time to half of peak intensity(HPT) delayed (P <0.05 or P <0.01). Compared with pre-injection, there were no differences in terms of the area under the curve(AUC) at 2,4 and 6 weeks after injection (all P> 0.05). And compared with pre-injection,2,4,6 weeks after injection, the AUC decreased at 8 weeks (P < 0.05 or P <0.01 ). The level of Scr and BUN of rabbits had increased since 6 weeks after injection (P< 0.05 or P <0.01). Two-dimensional ultrasound showed the renal volume was enlarged and the cortex was thickened from 2 weeks to 6 weeks after injection (P<0.05 or P <0.01). At 8 weeks after injection, the renal size had decreased as well (P < 0.05). These ultrasound changes were in accordance with its pathological changes. Conclusions Contrast-enhanced ultrasound in combination with time-intensity curve can quantitatively analyze the renal perfusion of CRF at different stages. The reduction of renal perfusion was earlier than the changes of routine laboratory indexes in rabbits with CRF. The haemodynamic changes of CRF rabbit were closely related with its pathological changes.

Objective To evaluate the usefulness of real-time three-dimensional transesophageal echocardiography(RT-3D-TEE)in cardiac valvular diseases.Methods Preoperative and postoperative RT-3D-TEE studies were performed in 32 patients who had undertaken cardiac valve repair or valvular replacement using Philips iE33 with X7-2 probe,and quantitative analysed the mitral structure in cases of mitral valve prolapse by using online QLAB7.0 quantitative analysis software.Results RT-3D-TEE can demonstrate the anatomicsl structure clearly,evaluate the function of cardiac valves precisely,and reveal details of type,position and extent of lesions vividly in surgical view.RT-3D-TEE helped correct misdiagnose in 2 cases,revised surgical plans in 3 cases,and guided implementation of remedial surgery in 1 case.Conclusions RT-3D-TEE can present images lively,and it can provide adequate information for preoperative diagnosis,aid the formulation of surgical plan and evaluate surgical effect.

BACKGROUND: It is difficult to in vitro isolate and culture the ovarian surface epithelium with high purity, strong vitality and stable biological characteristics. Tissue adherence and enzymatic digestion are commonly used for primary culture, but there are certain problems in cel col ection, cel viability and cel purity. OBJECTIVE: To establish a method for primary isolating, culturing and identifying human ovarian epithelium. METHODS: The ovarian surface epithelium was obtained with cel brush innovatively, and then the cells were isolated and purified with erythrocyte spal ation and differential adherence. The epidermal growth factor was added into the serum-free Dulbecco’s modified Eagle’s medium-F12 medium for cel culture. The cel morphology was observed under inverted microscope, and hematoxylin-eosin staining and immunocytochemical staining were used to identify the cells, then the growth curve was draw. RESULTS AND CONCLUSION: The ovarian surface epithelium became adherent after cultured for 24 hours, and reached fusion basical y after cultured for 6-12 days. The cells were polygonal or flat with strong transparency and refraction. The morphological characteristics of the cells were in line with those of the normal epithelial cells, and almost al the isolated cells could express the epithelial cells surface marker CK19. The cells could be passaged for 6-8 generations with wel growth and the cel growth curve was in “S” shape. The purity of the cells was more than 95%. The results suggest that cel brush method is simple to operate and can obtain a large amount of ovarian surface epithelium rapidly. The purity of the isolated cells can reach to 95% after treated with erythrocyte spal ation and differential adherence method and the cells are in stable growth.

AIM: To investigate the expression and effects of sialyl Lewis X (SLeX) on the invasion and migration of human hepatocellular carcinoma HepG2 cells.METHODS: The expression of α1,3-fucosyltransferase VII (FUT7) in HepG2 cells and L-02 cells was detected by RT-qPCR and Western blot.The SLeX expression in HepG2 cells and L-02 cells was determined by Western blot and immunocytochemical staining.The invasion and migration abilities of the treated cells were evaluated by Transwell assay.RESULTS: The expression of FUT7 and SLeX in the HepG2 cells, but not in the L-02 cells, was observed.The invasion rates of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L were significantly decreased as compared with control group (P<0.05).The migration ability of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L was also significantly reduced as compared with control group (P<0.05).The invasion rate and migratory cell number were significantly different between any 2 groups in the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L (P<0.05).CONCLUSION: HepG2 cells express SLeX.SLeX is closely related to the migration and invasion abilities of the HepG2 cells.

Objective To investigate the efficacy of Ganju Bingmei Pill combined with voice therapy in the treatment of vocal nodules .Methods A total of 72 patients with vocal nodules and 60 health people in our hospital were included .Patients were randomly divided into two groups :Group A (36 cases) with Ganju Bingmei Pill thera‐py ,group B (36 cases) with Ganju Bingmei Pill combined with voice therapy .Taking method of Ganju Bingmei Pill was three times a day ,2 tablets every time ,two weeks as one course of treatment with a total of four courses .Voice therapy in Group B was two weeks for one course with a total of four courses .The results were evaluated by voice handicap index ,voice acoustic analysis and laryngostroboscopy .Vocal nodules disappear or significantly reduce indi‐cated that the treatment was effective .Statistical analysis was performed using t - test and chi - square test .Results In group A ,the scores of T ,F ,P ,E were reduced compared with before treatment (P < 0 .05) ,however ,com‐pared with normal control group ,the score of F ,E ,T were still higher than those of normal group (P < 0 .05) .In group B ,the score of T ,F ,P ,E were reduced compared with before treatment (P< 0 .05) ,and compared with normal control group ,the score of F ,P ,E ,T were close to normal(P> 0 .05) .The VHI score in group B were significantly lower than those of in group A (P < 0 .05) .In Group A compared with before treatment ,jitter ,shimmer ,NHR were lower ,F0 was higher(P < 0 .05) ,however ,compared with the group of normal control ,F0 ,jitter ,shimmer , NHR were still statistically significant (P < 0 .05) .In group B with before treatment jitter ,shimmer ,NHR were lower ,F0 was higher (P< 0 .05) ,and jitter ,shimmer were already close to normal(P> 0 .05) .The F0 of group B after treatment was higher than group A ,jitter and shimmer were lower than group A after treatment (P< 0 .05) . After treatment ,there were 28 cases of group A (77 .78% )of patients with vocal nodules disappear or significantly reduce .There were 35 cases of group B(97 .22% ) of patients with vocal nodules disappear or significantly reduce , the effective rate between group A and group B was statistically significant (P< 0 .05) .Conclusion Ganju Bingmei Pill therapy and Ganju Bingmei Pill therapy combined with voice therapy all have curative effect for the treatment of vocal nodules however ,Ganju Bingmei Pill therapy combined with voice therapy is more efficient .

Objective: To investigate the physical and intellectual development and mutation characteristics of the phenylalanine hydroxylase (PAH) gene among 53 newborns with phenylketonuria (PKU), so as to provide insights into the management and genetic counseling of PKU Methods: The medical records of 54 children with definitive diagnosis of PKU and standardized therapy until 2 years at the Center for Neonatal Disease Screening of Shanxi Children' s Hospital from 2018 to 2021 were collected. Newborns' body weight and height developments were evaluated using the World Health Organization growth chart (2006 version), and the intellectual development was assessed using the national criteria of Development Behavior Assessment Scale among Children at Ages of 0 to 6 Years (WS/T 580-2017). The gene mutations were detected among neonates and their children, and the physical, intellectual developments and genetic characteristics of neonates with PKU were descriptively analyzed. Results: The 53 PKU cases included 29 male children and 24 female children, 36 cases with classic PKU and 17 cases with mild PKU, and 30 cases from rural areas and 23 cases from urban areas. The study subjects had a median age of 30 (10) d at initial therapy, and a mean blood phenylalanine concentration of (1 507±685) μmol/L at definitive diagnosis. There were 52 cases with normal height developments (98.11%), and all cases had normal weight and intellectual developments. The mean developmental functional quotient (DFQ) was significantly greater among urban children with PKU than among rural children [(94.92±8.57) vs. (87.65±6.57); t=-3.498, P=0.001], and the mean DFQ was significantly higher among children with mild PKU than among those with classic PKU [(95.55±8.76) vs. (88.57±7.11); t=-3.095, P=0.003]. There were 37 mutations detected in the PAH gene, which were mainly distributed in exons 3, 6, 7, 11, 12 and intron 4. Three high-frequency mutation sites were detected, including c.728G>A, c.611A>G and c.1197A>T, including three novel mutations (c.674C>G, c.1316-2A>C and c.1069T>C). Conclusions Following standardized treatment, the children with PKU have comparable physical and intellectual developments as compared to normal children. c.728G>A, c.611A>G and c.1197A>T were predominant mutations in the PAH gene among these 53 children with PKU, and three novel mutations were identified, including c.674C>G, c.1316-2A>C and c.1069T>C.

Objective To investigate treatment effects of lentivirus mediated RhoA short hairpin RNA(shRNA) on xenograft tumor of ovarian cancer in nude mice in vivo and the underlying mechanism.Methods Human ovarian cancer cell line HO8910 were inoculated to establish subcutaneous xenograft model of human ovarian cancer.Tumor-bearing nude mice were assigned randomizely to three groups:LentiRhoA-sh group,Lenti-negative control (NC) group and phosphate buffered saline (PBS) group.lentivirus mediated RhoA shRNA,negative control lentivirus and PBS were respectively injected in the three groups.Effects of treatment were observed by tumor growth curve,tumor volume,tumor weight,and tumor inhibition rate.Xenograft tissues and liver,spleen,lung,and renal tissues were examined by hematoxylin and eosin (HE) staining or were detected by streptavidin-perosidase (SP)immunochemical method.The changes of RhoA gene expression in xenograft tissues after lentivirus mediated RhoA shRNA treated were also detected by real-time qPCR,immunochemistry and Western blot assay.Cell apoptosis in xenograft tissues were examined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL) method and apoptotic index (AI) were counted.Results Compared with Lenti-NC group and PBS group,the growth speed of xenograft in Lenti-RhoA-sh group delayed significantly after injection 9 days(P ＜ 0.01).Tumor volume (338 ± 114) mm3 decreased significantly in the Lenti-RhoA-sh group when compared with those in Lenti-NC group(1190 ± 332)mm3 and PBS group (1101 ± 396) mm3 (P ＜ 0.01).Tumor weight (0.23±0.11)g decreased significantly in the Lenti-RhoA-sh group when compared with Lenti-NC group (0.79 ± 0.19)g and PBS group (0.74 ± 0.17)g (P ＜ 0.01).Real-time qPCR result shown that the expression of RhoA mRNA (0.30 ± 0.05) decreased significantly in the Lenti-RhoA-sh group compared with Lenti-NC group (0.95 ±0.06) and PBS group(1.00 ±0.11 ; P ＜0.01).Western blot result showed that the expression level of RhoA protein decreased significantly in the Lenti-RhoA-sh group (0.14 ± 0.06) compared with those in Lenti-NC group(0.78 ± 0.14) and PBS group (0.75 ± 0.13;P ＜ 0.01).TUNEL staining displayed that AI significantly increased in the Lenti-RhoA-sh group (20.9 ± 3.4) ％ compared with those in Lenti-NC group(5.2 ±±2.0)％ and PBS group(6.0 ±2.1)％ (P ＜0.01).Conclusion Lentivirus mediated RhoA shRNA may be effectively down-regulate of the expression of RhoA,inhibit the growth of subcutaneous xenograft tumor of ovarian cancer in nude mice by increasing the cell apoptosis.

ObjectiveTo analyze the characterstics of phenotype and genotype of multidrug resistant tuberculosis (MDR-TB) and extensively drug resistant tuberculosis (XDR-TB) by molecular line probe assay and liquid culture with MGIT960.MethodsGenoType MTBDR Kits were used for identifying the types of the first-line and second-line antituberculosis drug resistant genes partly and BD MGIT960 was used for detecting the chug susceptibility.Results( 1 ) Out of 94 MDR-TB strains,the rate of drug resistant to EMB,AMK,OFX and MFX by BD MGIT960 assay were 36.2%,17.0%,54.3% and 55.3%,respectively.Among these isolates,13 were extensively drug resistant tuberculosis (XDR-TB).(2) Compared with MGIT960,the concordance rate of GenoType MTBDRplus was 86.2% and 95.7% respectively.Taking MGIT960 results as reference,the sensitivity of GenoType MTBDRsl detecting the susceptibility of EMB,AMK,OFX and MFX to 94 isolates were 47.1%,81.3%,94.1%,94.2%,respectively.The specificity were 75.0%,98.7%,90.7%,92.9%,respectively.(3) Among the rpoB mutation categories,S531L accounts for most.MTB resistant to IFN caused by the mutation of katG chiefly and the S315T1 was in the majority.The gyrA mutation sites located at the ninety-fourth codon most.Out of 94 strains,23 were mixed with 2 kindsof Mycobacterium tuberculosis at least and 7 were undetectable mutations.Conclusion Among the M/XDR-TB,the strains resistant to INH,RFP,AMK,OFX and MFX were caused most by the mutation of katG,rpoB,rrs and gyrA,respectively.The relationship between EMB and embB was not so clear relatively.As a fast detecting drug susceptibility test kit,GenoType MTBDR possess good sensitivity and specificity.So,it could be as an assistant method to guide the therapy on clinic.

OBJECTIVETo investigate the role of β-catenin and caspase-3 in amentoflavone-induced apoptosis of human colorectal cancer SW480 cells.

METHODSMTT assay was used to detect the viability of SW480 cells exposed to amentoflavone, and flow cytometry was employed to assess the cell apoptosis. Western blotting was performed to determine the protein expressions of β-catenin and caspase-3 in the exposed cells.

RESULTSAmentoflavone dose-dependently inhibited the viability of SW480 cells, and a high concentration of amentoflavone (150 µmol/L) obviously induced apoptosis of the cells. Amentoflavone exposure caused significantly increased expression of caspase-3 and suppressed β-catenin expression in the cells.

CONCLUSIONAmentoflavone-induced apoptosis in SW480 human colorectal cancer cells is associated with altered expressions of β-catenin and caspase-3.

Apoptosis ; Biflavonoids ; pharmacology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Colorectal Neoplasms ; pathology ; Humans ; beta Catenin ; metabolism