OBJECTIVE To compare the liver toxicity of matrine and oxymatrine ,and to explore their toxic mechanism. METHODS Thirty ICR mice were randomly divided into normal control ,matrine 200 mg · kg-1 and oxymatrine 200 mg · kg-1 groups,10 mice per group. After single ig administration of corresponding drugs or water, animal mortality was calculated at the 15th day. The content of glutamic-pyruvic transami?nase(GPT),glutamic-oxalacetic transamin(GOT),alkaline phosphatase(ALP)and lactate dehydroge?nase (LDH) in serum were detected. Histopathological changes of the liver were examined by HE stain. The content of superoxide dismutase(SOD)and glutathione(GSH)in liver homogenates were detected by ELISA. Hepatocyte apoptosis was detected by Tunel stain. RESULTS The mortality rate of mice in two groups was 80% and 0,respectively. GPT,GOT and ALP contents of dead mice in matrine group were significantly higher than that in normal control group(P<0.05). In oxymatrine group,only the content of ALP was increased(P<0.05). Four of the eight dead mice in matrine group exhibited liver cell necrosis(P<0.05),while only 1/10 mice in oxymatrine group had a mild liver cell necrosis(P>0.05). The content of SOD and GSH of dead mice in matrine group was lower than that in control group(P<0.05,P<0.01). The content of GSH in oxymatrine group was also decreased(P<0.05). The apoptosis rate of liver cells in dead mice in matrine group was increased(P<0.05). CONCLUSION A large dose of matrine and oxymatrine can produce liver toxicity. At an equal dosage,the liver toxicity of matrine is significantly higher than that of oxymatrine. The toxic mechanism is related to oxidative stress and apoptosis.

ObjectiveThe aim of this study was to establish women previously undergoing cesarean section again puerperal ways.Methods170 pregnant women who had previously undergone 1 or 2 cesarean section were studied. ResultsAmong the 170 cases, 93 cases were trial of labor. The success was 81 cases, the success rate was 87%. One patient with preuterine scar ruptured. There was no maternal death. When the repeat cesarean group was compared with the trial of labor group, in the blood loss and after delivery febrile morbidity were significantly higher in women with repeat cesarean (P＜0.05). The Apgar score of newborns was no difference (P＞0.05). ConelusionWe should control the indications for cesarean section. A trial of labor after previous cesarean is safe, less damage and can be rceommended in the majority of cases. All eligible women would allowed to deliver vaginally after previous cesarean.

Objective To explore whether combined olmesartan angiotensin Ⅱ receptor type 1 blocker(ARB) and angiotensin-converting enzyme inhibitor(ACEI) temocapril have synergistic effect on reversal of left ventricular hypertrophy (LVH) in stroke-prone spontaneously hypertensive rats (SHRsp). Methods Fourty-four SHRsps and 11 Wistar-Kyoto rats(WKY) were divided randomly into 5 groups:WKY-control group, SHRsp-control group, SHRsp-olmesartan 10 mg/(kg?d)group, SHRsp-temocapril 10 mg/(kg?d)group, and SHRsp-Olmesartan 3 mg/(kg?d)+temocapril 3 mg/(kg?d) group for 6 weeks. Hearts weight were measured and histologically studied. The mRNA expression of angiotensin Ⅱ receptor type 1(AT1R) and integrin ?1 in myocardium was detected by RT-PCR. Results Olmesartan, temocapril and the their combination significantly reduced systolic blood pressure in a similar magnitude. Combination therapy was shown not greater effect in reversal of LVH than by olmesartan alone, although the effect by both of them was greater than temocapril monotherapy. The mRNA levels of AT1R and integrin ?1 in SHRsp were significantly decreased by treatment with olmesartan, temocapril, or combination therapy. Olmesartan and combination therapy result in greater decreases in expression of AT1R and integrin ?1 mRNA in myocardium than that by temocapril. Conclusion Compared with olmesartan alone, the combination of ARS and ACEI didn't show synergistic effect on reversal of left ventricular hypertrophy as were down-regulation of AT1R and suppression of integrin ?1 mRNA in myocardium.

OBJECTIVE:To construct the database of“The Time and Age Limit in the Use of Common Orally Adminis-tered Drugs”in order to guide the rational drug use in patients.METHODS:The frequently used instructions of common orally administered drugs in the hospital where the authors worked were collected,and then were classified and listed according to the elements in“The Time and Age Limit in the Use of Common Orally Administered Drugs”,and further summarized according to the basic knowledge in drug administration.RESULTS:Ante cibum drugs accounted for22.2%,post cibum drugs11.7%,either ante cibum or post cibumor or unmarked drugs59.4%,bedtime drugs6.7%,wholly swallowed drugs12.2%,chewed drugs1.7%,children restricted drugs32.2%,and the drugs taken once daily with the set time accounted for13.9%.CONCL_ USION:The construction of“The Time and Age Limit in the Use of Common Orally Administered Drugs”database can guide patients to use medicine rationally.

AIM: To investigate the effect of Gengnianle Granule on hypothalamic-pituitary-ovarian(HPO) axis in climacteric rats.METHODS: 12-15 months SD female rats were randomized into climacteric group and young control group was selected additionally.After being administrated for thirty days,the level of E_2、P、Te、FSH、LH in serum were examined by the radio-immunity method,the expression of GnRH in hypothalame,the expression of ER in hypothalame and pituitary appendage were examined by the immunohistochemical method. RESULTS: Compared with climacteric group,Gengnianle Granule could increase the level of E_2、P in serum(P

OBJECTIVE:To get to know medical staff's recognition of ADR monitoring in our city.METHODS:The concept,reporting consciousness,range and reasons for ADR reporting,reporting processes,professional organization of ADR and law consciousness of ADR etc.were investigated in400medical staff members through questionnaires on“Medical Staff's Recognition Degree to the Monitoring of ADR”.RESULTS:The medical staff in our city were basically clear about the range and the reasons for ADR reporting,but they lacked adequate recognition to the concept and the consciousness of ADR report?ing,they were not clear about the reporting processes and the organization of ADR and they lack the law consciousness of ADR reporting.CONCLUSION:It is necessary to carry out training on professional knowledge,laws and regulations of ADR among the medical staff so as to enhance their recognition to ADR monitoring and to make ADR monitoring as a self-conscious behavior.

BACKGROUND: Key point for subculture of human embryonic stem cells (hESCs) is to inhibit spontaneous differentiation and make sure totipotency of cells. Although mouse embryonic fibroblasts (MEF) or human foreskin fibroblasts (HFF) used as the feeder layer can maintain undifferentiated state of embryonic stem cells, cell clone is still imperfect and parallel arranged. OBJECTIVE: To establish mixed feeder layer of mouse embryonic fibroblasts plus human foreskin fibroblasts and to observe the hESCs growth.DESIGN: Multi-sample observation and comparison.SETTING: Medical Center of Reproduction, the Affiliated Hospital of Hainan Medical College. MATERIALS: This study was performed at the Medical Center of Reproduction, the Affiliated Hospital of Hainan Medical College from April 2006 to July 2007. Foreskin was derived from the children who underwent circumcision and came from Urinary Surgery, the Affiliated Hospital of Haihan Medical College. The children's family members provided the informed consent, and the experiment received confirmed consent from the local ethic committee. hESCs line HN-1 was separated from human blastula. Eleven 12.5-14.5-day-old fetal mice of clean grade were selected in this study. The experimental animals were disposed according to ethical criteria. METHODS: Heads, four extremities, and viscera were removed from fetal mice under anesthesia. Subsequently, cell suspension was prepared using routine trypsinase digestion and inoculated. When cells were cultured in confluent monolayer, some primary cells were frozen, processed with mitocin-C for 2.0-3.0 hours, and inoculated based on the density of 1×108 L-1 in gelatin-coated dish, I.e., MEF feeder layer. The HFF separation and culture and the preparation of HFF feeder layer were the same as above-mentioned processing. In addition, the two fibroblasts were respectively counted and mixed together according to the ratios of 1∶0, 3∶1, 1∶1, 1∶3, and 0:1. And then, the mixture was inoculated based on the density of 1×108 L-1 in gelatin-coated dish, I.e., mixed feeder layer. The growth of subcultured hESCs in vitro was observed in three different feeder layers, and hESCs in the mixed feeder layer underwent alkaline phosphatase test, OCT-4 expression immunohistochemical measurement, and OCT-4 and telomerase mRNA expression RT-PCR detection. Finally, differentiation in vitro of hESCs was observed after removing the feeder layer.MAIN OUTCOME MEASURES: ① Growth of hESCs in the three different feeder layers; ② Growth of hESCs in the mixed feeder layer based on different mixed ratios; ③ undifferentiated state of hESCs in the mixed feeder layer; ④ differentiation in vitro.RESULTS: ① hESCs clone in the MEF and HFF feeder layers was thin and flat and imperfect, but hESCs clone in the mixed feeder layer was perfect and thick and solid. Apparently, the clone form in the mixed feeder layer was superior to MEF and HFF feeder layers. ② When MEF and HFF was mixed together according to the ratio of 1∶1, hESCs grew in apparent accumulation; clone border was clear; eminentia was apparent and perfect. However, there were no changes according to the ratio of 1∶3. The ratio of 1∶1 was superior to the ratios of 1∶0, 3∶1, and 0∶1. ③ Alkaline phosphatase staining and OCT-4 antigen expression were strongly positive. Specific straps of OCT-4 and telomerase mRNA expression were observed at 200-300 bp and 300-400 bp, respectively. ④ Embryoid bodies were formed. hESCs could differentiate into multi-morphological cells after attachment.CONCLUSION: ① The mixed feeder layer may well support in vitro subculture of hESCs to acquire excellent clone form compared to MEF or HFF feeder layer. ② The mixture of MEF and HFF has excellent effect according to the ratio of 1∶1.

[Objective]To evaluate the role of mature cumulus cells from oocyte-cumulus complex(OCC)in in-vitro maturation(IVM)and establish a new culture technique which is convenient to carry out.[Methods]The cumulus cells of OCC were cut off and dispersed by 1 mL syringe.The cumulus cells were co-cultured with the immature oocytes retrieved from the COH cycles after they adherent to the bottom of the dish.The immature oocytes were experienced IVM procedures in different culture media.They were divided into 3 groups(the oocytes at germinal vesicle stage from one woman were allotted to the same group randomly).Group 1(solution A):basic culture medium+human follicular fluid(hFF);Group 2(solution B):solution A+ cumulus cells(OCC);Group 3(solution C):solution A+ OCC+ follicle stimulating hormone(FSH)+ epidermal growth factor (EGF).Then,the maturation rate,fertilization rate and formation rate of available embryo were observed.[Results]In 113 treatment cycles,298 immature oocytes were performed IVM with solution A,B,and C.The difference for 24 hour maturation rates among 3 groups wag statistically significant(A:45.2%,B:61.7%,C:78.2%,P＜0.05).There was no statistical difference for 25～48 hour maturation rates and normal fertilization rates of mature oocytes.The differences of cleavage rates and rescued embryo rates between group 1 and 2,group 1 and 3 were statistically significant(P＜0.05).The formation rates of available embryo showed an increasing trend from group 1,2,to 3.[Conclusion]After being dispersed by simply beat upon with syringe and adherent culture,the mature cumulus eells from mature OCCs in COH cycles,together with growth factors in the follicular fluid or extraneously supplemented,could promote the IVM of immature oocyte.

BACKGROUND: It has been found that vascular endothelial growth factor can induce the differentiation of bone marrow mesenchymal stem cells into endothelial cells, but can the vascular endothelial growth factor gene promote the differentiation of bone marrow mesenchymal stem cells into vascular endothelial cells in the damaged organ under the hypoxic environment? OBJECTIVE: To observe whether human bone marrow mesenchymal stem cells induced by vascular endothelial growth factor could differentiate into vascular endothelial cells under hypoxia. METHODS: The third passage of human bone marrow mesenchymal stem cells were cultured in vitro. Cells in the control group were cultured with conventional culture medium, while those in experimental group were cultured with adenovirus vector containing vascular endothelial growth factor in 5% O2. After 2 weeks of culture, morphological observation and surface-related molecular detection were performed. The levels of vascular endothelial growth factor and endothelial nitric oxide synthase were detected by ELISA. The expression of endothelin and prostacyclin was detected by RT-PCR and western blot assay. RESULTS AND CONCLUSION: (1) The number of cells in the control group was more than that in the experimental group. The cells in the control group were crowded and arranged irregularly, showing a fiber-like growth, while those in the experimental group were mostly triangular or polygonal, exhibiting a colony-like growth. (2) CD31 was negative in the control group, while CD105 was positive and the positive rate was 99.7%, indicating that the cells still showed the phenotype of bone marrow mesenchymal stem cells. The positive rate ofCD31 was significantly increased to 30.33% in the experimental group and the positive rate of CD105 expression was decreased to 58.11%, indicating a typical phenotype of endothelial cells. (3) Compared with the control group, the expression of endothelin, vascular endothelial growth factor and endothelial nitric oxide synthase increased significantly in the experimental group (P < 0.05), and the expression of prostacyclin decreased significantly (P < 0.05). All these findings indicate that vascular endothelial growth factor can promote the differentiation of human bone marrow mesenchymal stem cells into vascular endothelial cells under hypoxia.

BACKGROUND:Kalikrein 1 is an important component of the kalikrein-kinin system. Studies have shown that kalikrein can protect the cardiovascular system by promoting angiogenesis and inhibiting myocardial inflammation, but there is no report on its effect on inducing differentiation of stem cels. OBJECTIVE: To determine the transfection efficiency of kalikrein 1 adenoviral vector in rat bone mesenchymal stem cels. METHODS:Using adenovirus as a vector, the target gene kalikrein 1 was transfected into rat bone marrow mesenchymal stem cels. Fluorescence microscopy, MTT method and flow cytometry were used to investigate the effect of transfection and determine the optimal multiplicity of infection. RESULTS AND CONCLUSION:Adenovirus carrying kalikrein 1 was successfuly transfected into rat bone marrow mesenchymal stem cels. Results from flow cytometry showed that the transfection efficiency was associated with the multiplicity of infection. When the multiplicity of infection was 150, the transfection efficiency was 80.8%. MTT results showed that when the multiplicity of infection was 200, the cel growth was inhibited remarkably. These findings indicate that adenovirus-mediated kalikrein 1 can be successfuly transfected into rat bone marrow mesenchymal stem cels with the optimal multiplicity of infection=150.