Objective To investigate the effect of berberine on growth and apoptosis of human cervical cancer cell line HeLa and its possible mechanism.Methods The effect of berberine on growth of HeLa cells was studied by MTT assay.Apoptosis of HeLa cells exposed to berberine was observed by flow cytometry and DNA gel electrophoresis.The expression of Bax and Bcl-2 was studied by Western blotting analysis.Results Berberine markedly inhibited the proliferation of HeLa cells in a time-and dose-dependent manner.After incubation of HeLa cells with 20 and 40 mg/L berberine for 48 h,DNA Ladder can be observed.A typical "sub-G1 peak" was checked by flow cytometry.There was a very low rate of natural apoptosis(1.9?0.6)%,while in 5 mg/L berberine group,the apoptosis rate was(2.3?0.8)%.After exposing HeLa cells for 48 h to 20 and 40 mg/L berberine,the apoptosis rate reached(16.7?2.8)%(P

[ ABSTRACT] AIM:To investigate the effect of polysaccharide from Fructus corni ( PFC) on cardiomyocytes against hypoxia/reoxygenation ( H/R) injury and its possible relationship with ROS/PKC/p38 MAPK pathway.METHODS:Prima-ry cardiomyocytes were isolated from neonatal SD rats and randomly divided into normal group, H/R group, PFC (20 mg/L, 100 mg/L and 200 mg/L) preconditioning+H/R groups, chelerythrine+PFC (100 mg/L)+H/R group and SB203580+PFC (100 mg/L)+H/R group.The cell viability was measured by inverted microscopic observation.Apoptosis in the car-diomyocytes was detected by Hoechst 33258 staining and fluorescence microscopy.The levels of lactate dehydrogenase ( LDH) and superoxide dismutase ( SOD) in the cell culture supernatants, and the reactive oxygen species ( ROS) in the cells were also measured by microplate reader.The protein levels of PKC, p-p38 MAPK and HSP70 in the cells were detec-ted by Western blotting.RESULTS:Compared with normal group, the cell viability and beating frequency were decreased in H/R group.LDH and ROS contents, apoptotic rate and p-p38 MAPK level increased significantly (P<0.01).Compared with H/R group, PFC preconditioning increased beating frequency, SOD activity and the protein level of PKC and HSP70, and decreased ROS production, the protein level of p-p38 MAPK and cell apoptotic rate.However, the effect of PFC was in-hibited by chelerythrine or SB203580.CONCLUSION:PFC may protect cardiomyocytes from hypoxia/reoxygenation injury. Its mechanism is possibly involved in the inhibition of ROS via increasing the activity of SOD and the activation of PKC, and suppression of excessive activation of p38 MAPK.

AIM: To establish the drug release method of Compound Salvia Multi-component released Doublelayer Tablet and investigate its in vitro drug release behavior and the influencing factors of the drug release rate.METHODS: The method of evaluating in vitro release rate of double-layer tablet was established with salvianolic acid B and gindenoside Rg_1 as index,The influencing factors of the release of rapid release layer and sustained release layer of double-layer tablet and their in vitro drug release behavior were studied.RESULTS: The rapid release layer showed the quick releasing effect; The drug release curve of sustained release layer accorded with Ritger-Peppas equation; artificial gastric juice had significant effect on the release curve of gindenoside Rg_1 which in double-layer tablets,but had no significant effect on the release curve of salvianolic acid B.CONCLUSION: The evaluation of in vitro release rate of Compound Salvia Multi-component released Double-layer Tablet shows good properties of fast and sustained release and clinic application is achieved.

Objective To explore the relationship between histological chorioamnionitis(HC),fetal vasculitis(FV)and the morbidity of neonatal respiratory distress syndrome(RDS).Methods Three hundred and forty-seven cases of infants at the gestational ages of 28 +0 to 31 +6 weeks who were admitted to the Neo-natology Department of our hospital from October 2009 to June 2013 were analyzed retrospectively.They were divided into four groups according to the occurrence of HC and FV,namely,HC positive group and HC negative group,FV positive group and FV negative group.The patients in the HC positive group were further divided into FV positive group and FV negative group according to the occurrence of FV.The morbidity of RDS among above-mentioned groups was compared.Results The clinical characteristics including propor-tion of male,gestational age,birth weight,cesarean delivery,antenatal corticosteroid use,gestational hyperten-sion,gestational diabetes and cholestasis of pregnancy showed no statistically significant difference among all the groups(P 〉0.05).The incidence of RDS in the HC positive group(145 cases)was 49.6%(72 cases), which was significantly lower than that in the HC negative group(67.3%,126 /202,P 〈0.05).The incidence of RDS in FV positive group(64 cases)was 42.2%(27 cases),which was significantly lower than that in FV negative group(63.3%,179 /283,P 〈0.05)In the HC positive group(145 cases),the incidence of RDS in FV positive group (64 cases)was 42.2% (27 cases),and 55.6% (45 cases)in FV negative group (81 cases),which showed no significant difference(P ﹦0.01 ).Conclusion (1 )HC or FV can reduce the incidence rate of RDS in premature infants.(2)HC combined FV cannot furtherly reduce the incidence of RDS.

Objective To investigate the physical and mental development of small and appropriate for gestational age preterm infants in their early life. Methods This study recruited 220 preterm infants, who were discharged from our hospital and visited preterm following-up clinic at regular intervals from February 2009 to December 2012. All of those infants were divided into two groups based on whether their birth weight below 10th percentile for their gestational ages or not. Weights, lengths and head circumferences were measured up to seventh month age adjusted by gestational age. Meanwhile, mental tests were conducted by the professional staffs working on the children developmental assessment at their adjusted months of 5th, 6th or 7th. All of physical and mental scores were compared between the two groups. Results The SGA group was statistically less than the AGA group on the Z-score of weights from the ifrst to sixth month adjusted by gestational age (P<0.05). The difference disappeared on the seventh adjusted month age (P>0.05). The SGA group was statistically less than the AGA group on the Z-score of lengths from the ifrst to iffth month adjusted by gestational age (P<0.05). The difference disappeared on the sixth and seventh adjusted month age (P>0.05). The SGA group was statistically less than the AGA group on the Z-score of head circumferences from the ifrst to seventh month adjusted by gestational age (P<0.05). The SGA babies scored statistically less than the AGA babies with a mean development quotient score of 96.7 and 102.9, respectively (P<0.05). The scores of movement, cognitive, language in the SGA group were statistically less than those in the AGA group(P<0.05). Conclusions Preterm SGA could achieve satisfactory weight catch-up gain, with a decreasing difference from preterm AGA while they were getting older. But the length catch-up growth of preterm SGA seemed unsatisfactory with a big differece from preterm AGA. There was the worst catch-up on head circumference in those preterm SGA, backward in mental development, particularly in their movement, cognitive and language capacity.

Objective To evaluate the role of protein kinase C (PKC ) in reduction of mitochondrial injury during myocardial ischemia-reperfusion (I/R) by ischemic preconditioning in rats .Methods Forty male Sprague-Dawley rats ,aged 12-13 weeks ,weighing 280-320 g ,were randomly divided into 4 groups ( n=10 each) using a random number table:sham operation group (S group) ,I/R group ,ischemic preconditioning group (IP group) and PKC inhibitor chelerythrine group (C group) .Myocardial I/R was produced by 35 min occlusion of left anterior descending branch of coronary artery followed by 120 min reperfusion .Ischemic preconditioning was induced by 3 episodes of 5 min occlusion of left anterior descending branch at 5 min intervals before myocardial ischemia . Chelerythrine 1 mg/kg was injected intravenously via the caudal vein before ischemic preconditioning in group C . At 120 min of reperfusion ,the animals were sacrificed and the hearts were immediately removed .Mitochondrial suspension was prepared for determination of activities of succinate dehydrogenase (SDH ) , xanthine oxidase (XOD ) , glutathione peroxidase (GSH-Px ) and Ca2+-ATPase , content of Ca2+ , myocardial mitochonerial permeability transition pore (mPTP) opening and membrane potential (Δψm ) .Results Compared with S group , the activities of XOD and Ca2+-ATPase ,content of Ca2+ and mPTP opening were significantly increased ,and the activities of SDH and GSH-Px and Δψm were decreased in I/R group ( P<0.05) .Compared with I/R group ,the activities of XOD and Ca2+-ATPase , content of Ca2+ and mPTP opening were significantly decreased , and the activities of SDH and GSH-Px and Δψm were increased in IP group ( P<0.05) .Compared with IP group ,the activities of XOD and Ca2+-ATPase , content of Ca2+ and mPTP opening were significantly increased , and the activities of SDH and GSH-Px and Δψm were decreased in C group ( P<0.05) .Conclusion PKC is involved in reduction of mitochondrial injury during myocardial I/R by ischemic preconditioning in rats .

Objective To explore the correlation between histological chorioamnionitis (HC) and brain injury in preterm infants. Methods Three hundred and forty-seven cases of infants at the gestational age of 28-31 weeks who were admitted to the neonatology department of our hospital were analyzed retrospectively. They were divided into the HC group and the control group according to the pathological examination. Moreover, HC group was divided into FV group and non-FV group according to the pathological findings of fetal vasculitis (FV). Based on the findings of periodical ultrasonography, the incidences of periventricular leukomalacia (PVL), periventricular-intraventricular hemorrhage (PVH-IVH), and the PVL+PVH-IVH were compared among groups. Results The incidences of PVL in the HC group and the control group were 17.9% and 10.3%respectively. The incidences of PVL+PVH-IVH in the two groups were 5.5%and 1.48%respectively, and the difference between two groups was signiifcant (P<0.05). The incidences of PVH-IVH in the two groups were 28.9%and 26.2%respectively, and the difference between two groups was not signiifcant (P>0.05). In the HC group, the incidences of PVL in FV group and non-FV group were 28.1%and 9.87%respectively, and the difference between two groups was signiifcant (P<0.05). The incidences of PVH-IVH in FV group and non-FV group were 34.3%and 24.7%respectively, and the difference between two groups was not signiifcant (P>0.05). The incidences of PVL+PVH-IVH in FV group and non-FV group were 7.81%and 3.70%respectively, and the difference between the two groups was not have signiifcant (P>0.05). Conclusions HC may increase the ncidences of PVL and PVL+PVH-IVH in the preterm infants, while its effect is minimal on PVH-IVH. FV could increase the incidence of brain injury in preterm infants.

Objective To investigate the impacts of LASP-1 on the behavior and cell proliferation ability of tumor derived osteoblast-like cells and non tumor derived osteoblasts. Methods QRT-PCR and Western blot were performed to detect LASP-1 expressions in mouse bone marrow mesenchymal stem cells(BMMSC),human osteoblasts (hOB) and human osteosarcoma cell (MG-63). With the construction of recombinant vector and the transfection of shLASP-1,the expression changes in both LASP-1 and osteocalcin(OCN),and the cell number of cell-cycle phase distribution were analyzed. Results With up-or down-regulation of LASP-1,osteocalcin(OCN) expression,cell proliferation ability and the cell number of G2/M phase in non tumor cells(BMMSC,hOB)had no obvious changes. However,as for the tumor cells(MG-63),with an increased expression of LASP-1,the expression of OCN,cell proliferation number and the cell number at G2/M phase changed. There was no significant effect on the moving ability of each cell. Conclusions LASP-1 has no significant effect on the osteogenic proper-ties,cell proliferation ability and cell cycle distribution of non tumor derived cells,but has a regulatory effect on tumor derived cells(MG-63).

Objective To establish a noninvasive diagnostic model for liver fibrosis in chronic hepatitis B(CHB)patients with normal alanine aminotransferase(ALT)and an age of≤30 years by selecting specific indicators from the commonly used noninvasive indicators such as clinical,biochemical,and imaging indicators,to avoid invasive liver biopsy in such patients to some extent,and to guide the timing of antiviral therapy.Methods A total of 251 CHB patients with normal ALT and an age of≤30 years who underwent liver biopsy in Shenzhen Third People's Hospital and The First Hospital of Changsha from January 2019 to January 2022 were enrolled,with 175 patients in the model group and 76 patients in the validation group,and commonly used clinical indicators were obtained based on clinical experience and related articles.The two-independent-samples t test or the two-independent-samples Mann-Whitney U rank sum test was used for comparison of continuous data between groups,and the chi-square test was used for comparison of categorical data between groups.A Spearman rank correlation analysis was used to investigate the correlation between each indicator and liver fibrosis and identify the indicators with correlation(P<0.01,r>0.200);a Logistic regression analysis was used to establish a noninvasive diagnostic model,and the receiver operating characteristic(ROC)curve was used to evaluate its performance and perform validation of the model;this model was then compared with the widely used models of aspartate aminotransferase-to-platelet ratio index(APRI)and fibrosis-4(FIB-4).The Kappa consistency test was used to investigate the consistency of pathological results.Results A total of 17 commonly used clinical indicators were obtained,among which 9 indicators(ALT,aspartate aminotransferase[AST],gamma-glutamyl transpeptidase[GGT],ferritin[FERR],platelet count[PLT],procollagen type Ⅲ amino-terminal peptide[PⅢP],collagen Ⅳ[CⅣ],HBV DNA,and spleen thickness)were correlated with liver fibrosis(P<0.01,r>0.232).Based on the above indicators,the predictive model was established as P=1/(1+e-γ),γ=-1.902+0.106×AST-0.011×PLT-0.265×HBV DNA+0.059×PⅢP,in which P was the probability for predicting≥S2 liver fibrosis and γ was the predictive index.The comparison between each indicator and the model showed that the model had the largest area under the ROC curve of 0.852,with a sensitivity of 92.7%and a specificity of 76.9%.The model was validated in 76 patients and showed an accuracy of 77.600%.The model was compared with APRI and FIB-4,and the results showed that the model has good accuracy.Conclusion Compared with the models of APRI and FIB-4 commonly used in the world,this model can more accurately judge the degree of liver fibrosis in such patients,thereby replacing liver biopsy to some extent and guiding the timing of antiviral therapy.

OBJECTIVETo prepare enteric coated pellets containing panax notoginseng saponins.

METHODPanax notoginseng saponins loaded pellets were prepared by Extrusion-Spheronization method, and coated by Eudragit L30D-55 using Glatt fluid bed with the bottom spray process, central composite design was used to optimize the coating prescription.

RESULTThe drug release of enteric coated pellets of panax notoginseng saponins pellets would be lower than 5% in 2 h in simulated gastric fluid, but reach above 85% in 3 h in simulated human gastroenteric environment.

CONCLUSIONThe enteric coated pellets of panax notoginseng saponins have good acid residence to avoid panax notoginseng saponins from degrading in gastric acid.

Chemistry, Pharmaceutical ; methods ; Drug Stability ; Gastric Acid ; chemistry ; Gastrointestinal Tract ; drug effects ; Humans ; Models, Biological ; Panax notoginseng ; chemistry ; Saponins ; chemistry ; pharmacokinetics ; Tablets, Enteric-Coated ; chemistry