Objective To study whether solasodine hydrochloride (SBHL) could enhance the effect of arsenic trioxide in inducing apoptosis and affecting telomerase activity in cervical cancer HeLa cells. Methods Using cell culture methods, cervical cancer HeLa cells were cultured in vitro. The optimal concentration of SBHL was determined by MTT method from 0, 10, 20, 40, 80, 160, to 320 μmol/L. HeLa cells were grown in improved RPMI1640 supplemented respectively with arsenic trioxide(5 μmol/L As2O3), As2O3(5 μmol/L)+ SBHL( 40 μmol/L) and none (control group). The growth morphology of HeLa cells was observed under phase contrast microscopy after culture for 24, 48, and 72 h. Apoptosis of HeLa cells was determined under transmission electronic microscopy. The method of MTT was used to study the cell survival percentage. The technique of flow cytometry was used to measure cell cycle and cell apoptosis percentage. The method of tartrate-resistant acid phosphatase-enzyme linked immunosorbent assay (TRAP-ELISA) was used to determine telomerase activity of HeLa cells. Results Under phase contrast microscopy, in control group HeLa cells were round, densely packed; in As2O3 group the numbers of the cells were less, cell spacing increased; in As2O3 + SBHL group the cells shrinked significantly, nuclear fragmented as a petal-like, gap became larger. Under transmission electronic microscopy, there were rich microvillus on the cell surface in control group, cell intervals clear, immature connections, and the intervals did not close. The structure of the mitochondria in the cytoplasm was integrated. Most of the chromatin in the nucleus were, euchromatin and characteristics of apoptosis with heterochromatin increased and the chromatin condensed into masses, on the boundary of nuclear membrane. The microvillud on the cell surface were ruptured and decreased in As2O3 + SBHL group. The chromatin condensed into masses. The formation of apoptotic bodies was observed. The difference was statistically significant between groups in cell survival percentage at 24, 48, 72h(x2 = 10.39 , 13.88 , 17.21,respectively, all P < 0.05). Cell survival percentage in SBHL + As2O3 group (52.80%) was significantly less than that of As2O3 group(77.51%, x2 = 9.29, P < 0.05) at 72 h. In cell cycles, the difference was statistically significant between groups in C1 phase and S phase(F = 7.46,22.14, all P < 0.05), respectively. Compared with , control group[ (41.57 ± 1.56)%, (50.45 ± 2.37)%], cell percentages in S phase in As2O3 + SBHL group[(20.06 ± 4.98)%] and As2O3 group[(27.10 ± 5.32)%] were decreased(P< 0.05 or < 0.01), while cell percentage in C1 phase was increased[(58.70 ± 5.18)%, (69.67 ± 4.17)%, P< 0.05 or < 0.01]. The difference was statistically significant between groups in apoptotic percentage of HeLa cells (F = 4.01, P < 0.05). Compared with control group[ (1.18 ± 1.40)%], apoptosis percentage was significantly increased in As2O3 + SBHL group and As2O3 group [(21.08± 1.22)%, (6.04±2.53)%, P< 0.05 or < 0.01], respectively, and As2O3 + SBHL group was higher than As2O3 group(P < 0.01). The difference was statistically significant between groups in telomerase activity (F = 21.28, P< 0.05). Telomerase activity was inhibited in As2O3 group(1.214 ± 0.621) and As2O3A + SBHL group(0.865 ± 0.284) compared to control group (2.107 ± 0.057, all P < 0.05), and telomerase activity in As2O3 + SBHL group was lower than that of As2O3 group (P < 0.05). Conclusions SBHL enhances the effect of As2O3 in inducing apoptosis in HeLa cells, which is related to its inhibiting telomerase activity in HeLa cells.

Adult ; Humans ; Lead Poisoning ; etiology ; Male

ObjectiveTo observe testis tissue changes of experimental cryptorchidism after orchiopexy in various-day-rats.MethodsSeventy-two Sprague-Dawley(SD) male rats were divided into 3 groups randomly and made artificial cryptorchidism:unilateral cryptorchidism group(n=24),bilateral cryptorchidism group(n=24)and sham operation group(n=24)at age 21-day-old.Intra-abdominal testicle was resetting in 2 weeks,at age 40 days and 60 days,the rats were sacrificed for detection tubular fertility index(TFI) and mean tubular diametar(MTD) with hematoxylin-eosine staining and germ cell apoptosis by terminal deoxynucleotidyl transferase mediated d-UTP nick end labeling(TUNEL) assay.ResultsThere were significant differences of MTD,TFI and apoptosis index(AI) between cryptorchidism and scrotal testes(P0.05).The AI of cryptorchidism testes of unilateral cryptorchidism group were significantly lower than that of bilateral cryptorchidism group in 40 days(P0.05).ConclusionsAI of artificial resetting testis is increased and contralateral descended testes in unila-teral cryptorchidism have various damage.It is to lighten that pathological damage of testes of cryptorchidism with prolongation of reset time.

The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.

Objective To explore the optimal methods to detect helicobacter pylori(Hp)antigen in children.Methods 13C labeled urea breath test(13C-UBT)was performed on 937 children of alimentary department from Sep.2000 to Feb.2006.Gastric mucosa biopsy of 96 children was detected.Hp stool antigen(HpSA)status of the 557 children were evaluated.Both 13C-UBT and HpSA were assayed in 105 children from Apr.2003 to Apr.2004.Method of 13C-UBT was taken as the golden standard on diagnosis of Hp infection.Results Forty-one point seven percent children was positive for 13C-UBT.Forty point six percent children was positive for gastric mucosa biopsy.Thirty-eight point two percent was positive for HpSA.The difference was not significant.2.Among the 105 children performed by both 13C-UBT and HpSA,41.9% was positive for 13C-UBT and 39.0% for HpSA.Taking 13C-UBT as the golden standard,sensitivity of HpSA to diagnose Hp infection was 91.8%,and its specificity was 81.8%.Consistency Kappa coefficient was 0.743(P=0),which denoted that there was no significant difference on the positive detection rate between 13C-UBT and HpSA(P=0.388).Conclusions 13C-UBT and HpSA as non-invasive technique is effective to detect the Hp antigen.Compared to 13C-UBT,as a convenient,noninvasive,economical method,HpSA detection is much more acceptable to children and their patients.

OBJECTIVETo investigate the protective effect of extracts from Cichorium endivia (CEE) in H2O2-induced HepG2 cell oxidative stress injury, and explore the antioxidant mechanism of CEE in HepG2 cells.

METHODThe viability of H2O2-induced HepG2 cells and the intracellular ROS level were measured by MTT assay and DCFH-DA fluorescence staining assay. The antioxidant-response element (ARE)-Luciferase activity was tested in HepG2 cells stably transected by ARE reporter gene. The fluorescence quantitative RT-PCR was adopted to determine the mRNA expressions of genes containing ARE sequence in HepG2 cells.

RESULTThe cell viability reduced, while the ROS level increased after HepG2 cells were treated by H2O2. Different concentrations of CEE could be added to significantly improve the above results. After HepG2 cells transected by ARE reporter gene were treated with different concentrations of CEE, the intracellular ARE activity could increase in a concentration-dependent manner. In addition, the mRNA expressions of regulatory genesGCLC, GCLM and HMOX-1 containing ARE sequence in HepG2 cells were up-regulated in a concentration-dependent manner by CEE.

CONCLUSIONCEE inhibited the H2O2-injured HepG2 cells by reducing the ROS level. CEE's antioxidant mechanism for HepG2 cells may be closely related to the antioxidant defense system associated with its effect of activating Nrf2-ARE pathway in HepG2 cells.

Antioxidants ; isolation & purification ; pharmacology ; Asteraceae ; chemistry ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hep G2 Cells ; Humans ; Hydrogen Peroxide ; pharmacology ; Reactive Oxygen Species ; metabolism ; Response Elements ; genetics

In recent years, layer-by-layer self-assembly (LbL) has developed rapidly. It has been widely used in various industries such as medicine and metallurgy because of its simplicity, flexibility and controllability. In the study of drug delivery system, hollow microcapsules constructed by LbL method as drug carrier have great advantages in drug release, circulation in vivo and bioavailability, providing a technical platform for targeted drug release. In this paper, we summarize the types of film-forming materials and the driving force used in LbL technology, the way of loading drug into hollow micro capsule, and the variety of loaded drugs. We focus on the release mechanism, its evaluation and safety evaluation of self-assembled film as drug carrier in vivo and in vitro. The review shows the great application prospect of LbL technology in the field of drug delivery.

Objective To evaluate the value of three-dimensional contrast-enhanced angiography (3D CE MRA) and MRI in the diagnosis and delineation of cerebral arteriovenous malformation(AVM). Methods Twenty-two cases of cerebral AVM examined by MRI and 3D CE,MRA.DSA was performed in 17 cases.A three-dimensional fast low angle shot (3D FLASH) was used for 3D CE MRA with Gd-DTPA dosage of 0.2 mmol per kilogram for body weight.The source images were subtracted from mask images and transferred to computer workstation using three-dimensional reconstruction.Results Among 22 cases, 19 showed typical AVM“flow void”signal on MR images.3D CE MRA clearly displayed the nidus,feeding artery and draining vein.All of the foci were above the cerebellum tentorium.13 located within one lobe, 3 exceeded one lobe and 3 situated in the deep of cerebrum.Feeding arteries were derived from single artery in 9 cases,and mixed supply in 10 cases.Draining veins diverted to sagittal sinus and/or sigmoid sinus in 6,deep cerebral veins in 8 and mixed in 5.In the 14 cases examined by both DSA and 3D CE MRA,3D CE MRA was superior to DSA in three-dimensional demonstration of the nidus,but inferior to DSA in demonstration of some details.3D CE MRA depicted 78.4% feeding arteries and 84.0% draining veins in addition,tiny pathologic blood vessels smaller than 1 centimeter were detected by DSA,but could not be found by 3D CE MRA and were only shown as hemorrhage lesions on MR images in 3 cases.Conclusion As a non-invasive technique,3D CE MRA combined with MRI is accurate in diagnosis and localization of cerebral AVM,and should be used as the first choice for those clinically suspected of AVM.But DSA remains needed for demonstration of details and tiny AVM.

0.05),the mean FA on the left was higher than the right(t=1.912,P

OBJECTIVETo investigate hepatitis E virus (HEV) infection among pigs in Henan province.

METHODSA total of 623 swine sera, collected from 5 districts, were divided into two groups, under 3-month of age and over 3-month of age. They were tested for HEV antigen and antibody by using ELISAs, respectively. The sera positive for HEV antigen were tested for HEV RNA with RT-PCR. The positive products of RT-PCR were cloned and sequenced.

RESULTSThe positive rates of anti-HEV antibody of the groups under 3-month and over 3-month of age were 90.27% and 92.55%, respectively, without statistical difference, while those of HEV antigen were 15.93% and 5.69%, respectively, with significant difference. The positive rates of anti-HEV antibody and HEV antigen were significantly different among different districts. HEV RNA was detectable in 5 of 47 HEV antigen positive samples. The sequence analysis showed that in 4 of 5 specimens the sequence belonged to genotype 4 while in the remaining one the sequence was genotype 1.

CONCLUSIONThe prevalence rate of HEV infection in pigs was high in Henan province and the rate differed in different districts.

Animals ; Antibodies, Viral ; analysis ; immunology ; Antigens, Viral ; analysis ; immunology ; China ; Genotype ; Hepatitis E ; epidemiology ; immunology ; veterinary ; virology ; Hepatitis E virus ; genetics ; immunology ; isolation & purification ; Phylogeny ; RNA, Viral ; analysis ; genetics ; Sequence Analysis, DNA ; Swine ; virology ; Swine Diseases ; epidemiology ; immunology ; virology