AIM: To investigate the effect of mycophenolate acid(MPA) on hepatitis B virus(HBV) replication in vitro.METHODS: In the presence or absence of guanosine,the HepG2.2.15 cells were treated with different concentrations of MPA(1-20 mg/L) for 4 days.Hepatitis B surface antigen(HBsAg) and hepatitis Be antigen(HBeAg) in supernatant were detected by ELISA.Intracellular HBV core mRNA and HBV DNA were analyzed by reverse transcription-polymerase chain reaction(RT-PCR) and slot blot hybridization,respectively.RESULTS: MPA suppressed the expression of HBsAg and HBeAg,and inhibited the replication of HBV DNA.The effect of MPA on HBV replication was reversed by addition of exogenous guanosine.CONCLUSION: MPA suppresses the expression of HBsAg,HBeAg and replication of HBV DNA in HepG2.2.15 cells.Reducing the synthesis of guanosine nucleotides may be involved in the mechanism of the inhibitory activity of MPA on HBV replication.

Objective To observe the effect of topical application with warm and wet carthamus tinctorius alcohol and anisodaminum on phlebitis caused by mannitol injection. Methods 100 patients with fractured bones suffered from phlebitis caused by mannitol injection were randomized into the observation group and the control group with 50 cases in each group. The observation group adopted topical application with warm and wet carthamus tinctorius alcohol and anisodaminum while the control received external application of 50% magnesium sulfate. The dosage was 3 hours per time,2 times a day, one time in the morning and afternoon respectively. The treatment continued till the phlebitis disappeared and the effect at the 24th,48th and 72th post treatment was compared between the two groups. Results The effect at the above mentioned time points in the obsevation group was superior to that of the control group(P＜0.05).Conclusion Topical application with warm and wet carthamus tinctorius alcohol and anisodaminum on phlebitis caused by mannitol injection proved to be safe and had no adverse effect.

Objective To investigate the association of osteoporosis and coronary artery calcification in elderly patients with type 2 diabetes mellitus (T2DM).Methods A total of 82 elderly T2DM patients underwent dual-energy x-ray absorptiometry scanning (DXA) of lumbar spine and femur neck for getting bone mineral density (BMD),and dual-source computed tomography (DSCT) of coronary artery for calculating calcification score and total calcification score (TCS).All subjects were divided into two groups:osteoporosis group and non-osteoporosis group.The levels of serum calcium (Ca),parathyrin (PTH),phosphorus (P),alkaline phosphatase (AKP),triglyceride (TG),total cholesterol (TC),high density lipoprotein-cholesterol (HDL-C),low density lipoproteincholesterol (LDL-C) and glycosylated hemoglobin (HbA1c) were detected.Results Compared with non-osteoporosis group,the levels of serum Ca,PTH and TCS were higher [(2.32± 0.15)mmol/L vs.(2.04±0.20) mmol/L;(5.64±1.97) pmol/L vs.(5.01±1.93) pmol/L;(374.4±433.5) scores vs.(242.5±224.8) scores,t=5.790,5.331 and 2.248,all P<0.05] in osteoporosis group.Correlation analysis showed TCS was negatively associated with BMD of L2-4 and femur neck,while was positively associated with serum Ca and PTH (r=0.310,0.246,0.290,0.284 and 0.324,0.575 all P<0.05).Conclusions Osteoporosis is associated with coronary atherosclerosis.TCS could be considered as an index for judging the relationship between osteoporosis and coronary atherosclerosis.

AIM: To investigate lymphotactin (Lptn) gene transcription during acute cardiac allograft rejection and the inhibitory effect of cyclosporine A (CsA). METHODS: Graft specimens were harvested at indicated time to determine morphological changes by pathological examination. The grade of acute cardiac allograft rejection was evaluated by using modified Banff scoring system. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the Lptn mRNA expression in cardiac grafts. NFATc1 activity of splenocytes after transplantation was assessed by enzyme-linked immunoabsordent assay (ELISA). RESULTS: Prominent splenomegaly on day 3 posttransplantation was found in C57BL/6-Balb/c group. The extent of myocardial inflammatory infiltration was scored 2.667?0.577 at day 5 and 2.333?0.577 at day 7, respectively. Splenomegaly was ameliorated by CsA treatment, and the extent of myocardial infiltrate was scored 1.000?0.000 at day 5 and 1.333?0.577 at day 7, respectively. Lymphotactin mRNA was undetectable in cardiac isografts. Lymphotactin mRNA, which was inhibited partially by CsA, was upregulated strongly in acutely rejecting cardiac allografts at day 5 and day 7. Further studies demonstrated that NFATc1 activity in splenocytes, which markedly upregulated during acute rejection, was completely inhibited by CsA. CONCLUSION: Lptn appears to be a key chemokine of lymphocyte infiltration during acute allograft rejection. Inhibition of NFATc1 activity by CsA seems to decrease Lptn expression incompletely, suggesting that there was else mechanism to regulate Lptn expression other than NFAT pathway.

The cortexes were obtained from new-born rats and dissociated to single cells by triturating. The cells were cultured in neural stem cell (NSC) culture medium (DMEM supplemented with bFGF, EGF and B27) and formed primary neurospheres after 7 days. Single cells dissociated from neurosphere were cultured in 96-well plates and formed single-cell cloning neurosphere 7 days later. The primary and single-cell cloning neurospheres were both positive for the immunofluorescent staining of nestin and were identified as NSC. It was proved that NSC can be expanded in vitro and provide seed cells for neural tissue engineering.
Animals, Newborn ; Cell Separation ; Cells, Cultured ; Cerebral Cortex/*cytology ; Culture Media ; Neurons/*cytology ; Rats, Sprague-Dawley ; Stem Cells/*cytology ; Tissue Engineering

Objective To investigate the change of adiponectin level with menopause status in women aged 40 to 65, and its relationship with androgen. Methods A cross-sectional study included woman (aged from 40 to 65) who were in hospital for routine check-up at the Sun Yat-sen Memorial Hospital from August to October in 2013. All subjects underwent laboratory examinations of adiponectin, sex hormone binding globulin (SHBG), dehydroepiandrosterone-sulfate (DHEA-S), total testosterone (TT), collected anthropometric measurements and then calculated free androgen index (FAI) and body mass index (BMI). According to their menstrual status, the subjects were divided into 4 groups: premenopausal group with 119 subjects, perimenopausal group with 60 subjects, early postmenopausal group with 62 subjects, late postmenopausal group with 64 subjects. Results (1) Adiponectin levels declined to its lowest level in menopausal transition and gradually becoming higher after menopause, which showed a U-shaped trajectory. When compared adiponectin levels in late postmenopausal group [(13 ± 5) mg/L] with those in perimenopausal [(8 ± 6) mg/L] or early postmenopausal group [(9 ± 6) mg/L], it all showed significantly difference (P<0.05). (2) Both the adiponectin levels were negatively correlated with waistline in the 4 groups (premenopausal group, r=-0.276;perimenopausal group, r=-0.334;early postmenopausal group, r=-0.211;late postmenopausal group, r=-0.218; all P<0.05). Levels of adiponectin were positively correlated with SHBG (r=0.536, P<0.05) and negatively with FAI (r=-0.363, P<0.05) in menopausal transition, while in late postmenopausal group, negatively correlated with level of DHEA-S (r=-0.450, P<0.05). When adjusted for age, BMI and waistline, the above correlations still exist. Conclusions Adiponectin levels declined to its lowest level in menopausal transition and gradually becoming higher after menopause, which showed a U-shaped trajectory during the sequential menopause status transition in middle aged women. Low level of adiponectin in menopausal transition is closely associated with the relative excess androgen occurred during this stage.

Objective To observe theRuangan granule on liver fibrosis in rats liver pathology change, the influence of hepatic function and hepatic fibrosis indexes, and to discusses the mechanism of its action to provide the basis for clinical prevention and treatment of liver fibrosis.Methods A total of 105 Wistar rats were randomly divided into a normal control group, a model group and a colchicines group, and Dahuang-Zhechong pill group, high-, medium- and low-doseRuangan granule groups (n=15 in each group). Liver fibrosis was induced by carbon tetrachloride and a high-cholesterol diet. After modeling, the low-, medium- and high-doseRuangan granule groups were intragastric administratedRuangan granule mixed suspension 3.6, 7.2, 14.4 g/(kg?d), respectively;Dahuang-Zhechong pill group was administrated with Dahuang-Zhechong pellets mixed suspension of 0.18 g/(kg?d); the colchicine group was intragastric administrated with colchicine mixed suspension of 0.108 mg/(kg?d); and the normal control group and the model group were intragastric administrated with the equal volume of distilled water. All rats were intragastric administrated for 8 weeks. HE staining and Masson trichromatic collagen staining were used to observe the pathological changes of liver While the change of AST, ALT, PH, TP and serum HA, LN, C-Ⅳ, PCⅢin blood serum were detected. Results Masson trichromatic collagen staining showed that, the percentage of liver collagen fiber area in rats of theRuangan granule high-dose group was significantly decreased (7.06 ± 1.18) % compared with model group (23.49 ± 1.34) %, colchicine group (11.35 ± 1.83) %, rhubarb worm pill group (15.27 ± 1.22) %,Ruangan granule medium-dose group (14.52 ± 1.75) %, and low dose group (16.08 ± 1.56) % (P<0.05 orP<0.01). Compared with model group,Ruangan granule high-dose group rats serum AST (75.86 ± 5.23 U/Lvs. 157.62 ± 24.04) U/L, the ALT (80.15 ± 5.94 U/Lvs. 160.58 ± 26.47) U/L, PH (52.58 ± 4.98μg/Lvs. 98.66 ± 6.75)μg/L significantly reduced, TP (74.19 ± 3.56 g/Lvs. 51.73 ± 5.92)g/L increased significantly (P<0.01).Ruangan granule high-dose group rats serum HA (277.22 ± 106.34 ng/mlvs. 553.19 ± 172.38 ng/ml), LN (89.82 ± 5.68 ng/mlvs. 134.25 ± 10.64 ng/ml), C-Ⅳ (47.94 ± 8.65 ng/mlvs. 84.18 ± 13.83 ng/ml), PCⅢ (16.53 ± 4.88 ng/mlvs.31.57 ± 5.35 ng/ml) decreased significantly in the model group (P<0.01).ConclusionRuangan granule has obvious effects for resisting liver fibrosis.

Objective To investigate the chemical constituents of Dimocarpus longan seeds in Sapindaceae.Methods The chemical constituents were isolated from the ethanol extract of D.longan seeds by silica gel column chromatography.Their structures were identified on the basis of physical and chemical properties and spectral analysis.Results One compound was isolated and identified as 2-methyl-1,10-undecanediol,named longandiol(1).Conclusion Compound 1 is a new compound.

Objective To investigate the effects of Ruangan granule on transforming growth factor-β1(TGF-β1)/Smads signaling pathway in liver fibrosis in rats. Methods A total of 105 Wistar rats were randomly divided into normal control group, model group and colchicine, Dahuang-Zhechong pill group, high-, medium- and low-dose Ruangan granule groups (n=15 in each group). Liver fibrosis was induced by carbon tetrachloride and a high-cholesterol diet. After modeling, the low-, medium- and high-dose Ruangan granule groups were intragastric administrated Ruangan granule mixed suspension 3.6, 7.2, 14.4 g/(kg?d), respectively;Dahuang-Zhechong pill group was administrated with Dahuang-Zhechong pellets mixed suspension of 0.18 g/(kg?d);the colchicine group was intragastric administrated with colchicine mixed suspension of 0.108 mg/(kg?d);and the normal control group and the model group were intragastric administrated with the equal volume of distilled water. All rats were intragastric administrated for 8 weeks. The expressions of TGF-β1, Smad3 and Smad7 proteins in the liver tissue were detected with immunohistochemical staining method. The expressions of TGF-β1, Smad3, Smad7 mRNAs in the liver tussue were detected by RT-PCR. Results The expressions of TGF-β1 (2.59 ± 0.99 vs. 0.43 ± 0.21) and Smad3 (2.56 ± 0.67 vs. 0.41 ± 0.18) proteins and TGF-β1 mRNA (2.25 ± 0.21 vs. 0.71 ± 0.09) and Smad3 (2.34 ± 0.03 vs. 0.78 ± 0.12) mRNAs in the model group were significantly increased than those in the normal control group (all P<0.01). Compared with the model group, the expressions of TGF-β1 (1.12 ± 0.27 vs. 2.59 ± 0.99) and Smad3 (1.05 ± 0.34 vs. 2.56 ± 0.67) proteins in the high-dose Ruangan granule group decreased significantly, the expression of Smad7 increased significantly (2.33 ± 0.62 vs. 0.36 ± 0.18), and the expressions of TGF-β1 (1.09 ± 0.11 vs. 2.25 ± 0.21) and Smad3 (1.10 ± 0.02 vs. 2.34 ± 0.03) mRNAs decreased significantly, the expression of smad7 mRNA (1.18 ± 0.13 vs. 0.38 ± 0.11) increased significantly (P<0.05). Conclusions Ruangan granule can regulate the TGF-β1/Smads signaling pathway via down-regulation of TGF-β1, Smad3 and up-regulation of Smad7 in liver fibrosis in rats.

Objective To investigate molecular diagnostic method for hereditarymethemoblobinemia. Methods The cDNA coding sequence of NADH-cytochrome b5 reductase (b5R) from 3 patients with hereditary methemoglobinemia was analyzed by direct sequencing of RT-PCR products and the genomic DNA of b5R gene by PCR-restriction endonuclease digestion or PCR-sequencing. Results The b5R cDNA of patient A was T/C heterozygous at nucleotide 527 and G/A heterozygous at nucleotide 608. The b5R cDNA of patient B was G/A heterozygous at both nucleotide 170 and nucleotide 179. The b5R cDNA of patient C was G/A heterozygous at nucleotide 608 and C/T heterozygous at nucleotide 791. Result of genomic DNA analysis was in agreement with that of cDNA approach. Conclusion The method for molecular diagnosis of hereditary methemoglobinemia was established and 3 novel b5R gene mutations were identified in compound heterozygosity in 3 Chinese patients.