ObjectiveTo observe and explore the effects ofJiakangning Capsules on the expression of ERK1/2 gene and proliferation of FRTL-5 by studying the effects ofJiakangning Capsules collaborated with siRNA on interfering ERK1/2 gene in FRTL-5.Methods FRTL-5 cells in good conditions were divided into control group, negative control group,Jiakangning Capsules collaborated with siRNA group, siRNA interference group, and Jiakangning Capsules group. RT-PCR was performed to detect the expression of ERK1/2 gene in mRNA levels in FRTL-5; Western blotting was performed to detect the expressions of ERK1/2 and p-ERK1/2; CCK8 method was used to detect cell proliferation.Results RT-PCR results showed that the ERK1/2 mRNA expression of the control group and negative control group were of no significant difference (P>0.05); compared with the control group, siRNA interference group andJiakangning Capsules group could inhibit ERK1/2 gene mRNA expression in FRTL-5 (P<0.01). Compared with the control group and negative control group, the ERK1/2 mRNA expression of Jiakangning Capsules collaborated with siRNA group decreased obviously (P<0.01). Compared with the control group and negative control group, the expression of p-ERK1/2, ERK1/2 ofJiakangning Capsules group, Jiakangning Capsules collaborated with siRNA group, and siRNA interference group decreased obviously (P<0.01). At the time of 24 h and 48 h after transfection, according to the results of CCK8, compared with the control group and negative control group, the cell proliferation ofJiakangning Capsules group, Jiakangning Capsules collaborated with siRNA group, and siRNA interference group was inhibited (P<0.01).Conclusion Jiakangning Capsules have blocking effect on the phosphorylation of ERK1/2. After ERK1/2 gene was silenced, the thyroid cell proliferation was inhibited. Jiakangning Capsules can collaborate with ERK1/2-siRNA to inhibit FRTL-5 cell proliferation.

Objective:To observe clinical therapeutic effect of Fu Fang Qi Dan Dai Zhu San on sequelae of cerebral hemorrhage. Methods: 56 cases were selected randomly from the 315 cases who had received functional exercise, massage and other rehabilitation treatments. The patient were administrated by Fu Fang Qi Dan Dai Zhu San Decoction for 3 months, which is constituted by Bu Yang Huan Wu Decoction and Fu Fang Dan Shen Tablets, and the therapeutic effect was compared with that of Bu Yang Huan Wu Decoction, Fu Fang Dan Shen Tablets and other clinically commomly - used drugs, respectively. Results: Both the clinically cured rate and the total markedly effective rate in the treatment group of Fu Fang Qi Dan Dai Zhu San were significantly higher than those of the six control groups. Conclusion: Fu Fang Qi Dan Dain Zhu San can obviously increase the therapeutic effect and shorten the therapeutic course, and it is a good prescription for treatment of sequelae of cerebral hemorrhage.

Objective To investigate the differentiation potential of human lung fibroblasts.Methods The study was performed in First Hospital of Tsinghua University from March 2006 to October 2006.Human lung fibroblasts were cultured for 2 weeks in an osteogenic medium[containing 10% fetal calf serum (FCS)and 50 g/mL Vitamin C and 10 mmol/L ?-glycerophospate],adipogenic medium(containing 15% horse serum,10-8 mol/L dexmethasone and 10 mg/L insulin),or control medium(10% FCS).Osteoblasts were detected by alkaline phosphatese and calcium salt staining.The expression of osteopontin was measured by Western blotting,Oil Red -O staining for identification of mature adipocytes.Results With the induction of osteogenic medium for 2 weeks,the expression of alkaline phosphatese and osteopontin was increased and the deposition of calcium salt was detected.Mature adipocytes formed after culture with adipogenic medium for 2 weeks.Conclusion Under certain condition,human lung fibroblasts can differentiate into osteoblasts or adipocytes,which were characterized by bone marrow mesenchymal stem cells.

BACKGROUND: Lung fibroblasts are believed to play an important role in lung tissue repair and regenerative process. The differentiation potential of lung fibroblasts is not known very well.OBJECTIVE: To investigate the multi-differentiation capacity of human lung fibroblasts.DESIGN: An observational comparative experiment.SETTING: Department of Respiratory Medicine, Beijing Shijitan Hospital and Central Laboratory of the First Hospital of Tsinghua University.MATERIALS: This study was performed at the Central Laboratory of the First Hospital of Tsinghua University from March 2006 to October 2006. Human adult lung fibroblasts were isolated as primary cultures from resected lung of patients with lung cancer.The study was approved by hospital's Medical Ethics Committee, and informed consent was signed. Dulbecco's modified Eagle's medium (DMEM), fetal calf serum (FCS), trypsin- ethylenediamine tetraacetic acid (EDTA), naphthol AS-MX phosphate and Fast Red TR were purchased from Sigma, USA. Mouse anti-human osteopondn antibody was from R&D Systems China Co., Ltd,Polyvinyl difluoride (PVDF) membranes were from Bio-Rad Laboratories Inc, Hercules, CA.METHODS: Human adult lung fibroblasts were isolated as primary cultures. Human lung fibroblasts were cultured in an osteogenic medium (containing 109-10-5 mol/L dexamethasone, 50 mg/L vitamin C and 10 mmol/L β-glycerophospate) and adipogenic medium (containing 15% horseurm, 10-8 mol/L dexamethasone and 10 mg/L insulin), or control medium (10% FCS).Ostcoblasts were detected by alkaline phosphatese (ALP) and calcium salt staining. The expression of osteopontin was measured by Western blotting. Oil Red-O staining was used for identification of mature adipocytes.MAIN OUTCOME MEASURES: The differentiation of lung fibroblasts induced by osteogenie medium; The differentiation of lung fibroblasts induced by adipogenic medium.RESULTS: With the induction of ostcogenic medium for 2 weeks, the differentiation of lung fibroblasts was induced by osteogenic and adipogenic medium, respectively. The expression of ALP and osteopontin was increased and the deposition of calcium salt was detected. After lung fibroblasts were cultured in adipogenic medium for 14 days, part of the cells gradually experienced morphological change from original spindle into oval shape. Oil Red -O staining indicated lipid drops accumulation within cytoplasm.CONCLUSION: Under certain condition, human long fibroblasts could differentiate into osteoblasts or adipocytes that were characterized by bone marrow mesenchymal stem cells.

Obective To evaluate the effects of dihydrotestosterone (DHT) on the expression of sterol regulatory element-binding protein-1c (SREBP-1c) in human HaCaT keratinocytes.Methods HaCaT cells were cultured in vitro and classified into 4 groups,i.e.,control group receiving no treatment,DIIT group treated with 3 different concentrations (10,100,1000 nmol/L) of DHT,LY294002 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the PI3K inhibitor LY294002 of 50 μmol/L,PD98059 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the MEK inhibitor PD98059 of 50 μmol/L.After another 24-hour culture,real time PCR and Western blot were carried out to detect the expression of SREBP-1c mRNA and protein in HaCaT cells,respectively.Western blot was also performed to determine the phosphorylation levels of protein kinase B (AKT),extracellular signal-regulated kinase (ERK),p38 mitogen activated protein kinase and c-Jun N-terminal kinase (JNK) in the HaCaT cells.Results DHT could enhance the expression of SREBP-1c mRNA and protein in HaCaT cells in a concentration-dependent manner,and induce the phosphorylation of AKT and ERK,but not that of P38 or JNK.The expressions of SREBP-1c mRNA and protein were significantly decreased in HaCaT cells treated with LY294002 plus DHT (7.4780 ± 1.2638 vs.21.6170 ± 2.2759,t =9.406,P ＜ 0.05; 0.7113 + 0.0313 vs.2.2577 + 0.0601,t =39.498,P ＜ 0.05),but experienced no statistical changes in those treated with PD98059 and DHT(both P ＞ 0.05),compared with those treated with DHT only.Conclusion DHT can induce the expression of SREBP-1c mRNA and protein in HaCaT cells,likely via the PI3K/AKT signaling pathway.

AIM: To investigate effect of Wujibaifeng Pills (WJBFP) on osteoporosis of ovariectonmized (OVX) rat. METHODS: Ovariectonmized (OVX) rat model was established to evaluate osteoporosis of which parameters investigated included bone gla protein (BGP), alkaline phosphatase (ALP), bone minera density (BMD), Serum phosphorus and serum total calcium. RESULTS: WJBFP(1.0g/kg,2.0g/kg,4.0g/kg) could enhance the contents of serum estradiol and calcitonin, decrease serum BGP level in OVX rats; It had no effect on serum total calcium and ALP activities but increase level of serum phosphorus; It could enhance BMD, prevent OVX rat from decreasing bone loss without raising body weight; furthermore, it could inhibit both the uterus and adrenal gland atrophy. CONCLUSION: WJBFP might have better prevention on osteoporosis of ovariectionmized rats.

Objective To discuss the effective early postoperative fluid management in infant living-donor liver transplantation. Methods From January 2008 to March 2009, 17 cases of infant living-donor liver transplantation were carried out. According to infant postoperative physiological and hemodynamic characteristics, rehydration was properly controlled on the surgery day, the negative fluid balance was achieved as soon as possible in three days. The infant condition changes, vital signs and urine output were closely monitored. At the same time, the nature and volume of fluid infusion was timely adjusted to maintain the infant in a stable environment in accordance with the laboratory tests. Results The early postopera-donortive hemodynamic stability was effectively maintained in 17 cases of infants, no case appeared with the capacity-related complications. Conclusions It is the key to reduce postoperative complications and mortality with effective circulating blood volume and hemodynamic stability and the negative fluid balance state in the postop-erative 3 days.

Objective To investigate the effect of high mobility group box chromosomal protein 1 (HMGB1) knockdown on improving renal function and decreasing cell proliferation of glomeruli in lupus nephritis (LN) MRL/Faslpr mice.Methods Twenty-four MRL/Faslpr mice were randomly divided into 3 groups:LN model group,shHMGB1 group and empty plasmid group.Besides,eight MRL/MpJ mice,age and mass matched to the MRL/Faslpr mice,were chosen as normal control group (shNC group).Electroporation technology was used for in vivo transfection in treatment group.shHMGB1 group and empty plasmid group were transfected by electroporation technology for shHMGB1 plasmids and empty plasmid,LN model group and normal control group were transfected only with saline.Automatic biochemical analyzer was used to detect serum urea nitrogen (BUN) and creatinine (Scr) levels and 24 h urinary protein (UP) was tested.HE staining was used to detect the pathological change of renal tissues; real-time PCR,immunofluorence staining and Western blotting were used to detect the mRNA and protein expression of HMGB1 and PCNA.Results (1) The HMGB1 mRNA and protein expression in LN group increased compared with those in control group,HMGB1 mRNA and protein expression in shHMGB1 group reduced compared with those in LN model group (all P ＜ 0.05).(2) 24 h UP of MRL/Faslpr mice in shHMGB1 group significantly reduced compared with those in LN group (P ＜ 0.05).(3) Immunofluorence and Western blotting showed that positive signal of proliferating cell nuclear antigen (PCNA) was mainly located in nuclei,PCNA mRNA and protein in glomeruli of LN model group increased compared with those of control mice (P ＜ 0.05).Interestingly,PCNA expression in glomeruli of shHMGB1 group remarkably reduced (P ＜ 0.05).Conclusions shHMGB1 significantly improves renal function and decreases cell proliferation of glomeruli in LN MRL/Faslpr mice.

Objective To explore the role and their relationship of Ki-67, VEGF and p27 expressed in adult patients with acute leukemia. Methods The expression of Ki-67, VEGF and p27 in bone marrow mononuclear cells (BMMNC) were analyzed by immunocytochemical staining, and their correlations of Ki-67, VEGF and p27 were statistically analyzed. Results The expression of Ki-67(42.48±25.78)% or VEGF (44.89±24.01)% on BMMNC from acute leukemia cells was significantly higher than that in the control (11.40±9.94)% or (16.90±12.54)% (P<0.01). But the expression of p27 (23.65±13.30)% was significantly lower than that in the control (50.23±22.68)% (P<0.01). The expressions of Ki-67 were positively correlated with and VEGF in patients with acute leukemia were positively correlated(r=-0.666, P<0.01), and the expressions of Ki-67 and p27 were negatively correlated with p27 in patients with acute leukemia (r=-0.316, P<0.05).Conclusion The evaluation of expression of Ki-67, VEGF and p27 on acute leukemic cells provides new insights to the pathogenesis is helpful in mechanism and is helpful in the diagnosis of acute leukemia.

OBJECTIVE:To study the pharmacokinetics effects of naloxone combination on Shenmai injection in rats in vivo. METHODS:12 rats were randomly divided into monotherapy group (Shenmai injection 9.00 ml/kg,iv) and combination group (Shenmai injection 9.00 ml/kg+naloxone 1.80 ml/kg,iv). The blood samples were collected before administration and 0.083,0.25, 0.5,0.75,1,1.5,2,3,6,12,24,48,96 and 144 h after administration. HPLC was adopted to determine the plasma concentra-tions of ginsenosides Rg1,Re and Rb1,and DAS 2.0 software was used to calculate the pharmacokinetic parameters. RESULTS:Compared with monotherapy group,the plasma concentration of ginsenosides Rg1 in combination group was increased,CL was de-creased,t1/2 and MRT were prolonged,and AUC0-144 h was increased;the plasma concentration of ginsenosides Re was increased,Ke was decreased,t1/2 was prolonged,MRT was shortened,and AUC0-144 h was increased;the plasma concentration of ginsenosides Rb1 was decreased,Ke was increased,t1/2 and MRT were shortened,and AUC0-144 h was decreased,with significant differences(P<0.01 or P<0.05). CONCLUSIONS:Shenmai injection combined with naloxone can slow down the removing of ginsenosides Rg1 and Re in vivo,and obviously the plasma concentration of Shenmai injection is higher than monotherapy group;speed up the removing of ginsenosides Rb1,and the plasma concentration of Shenmai injection is lower than monotherapy group obviously.