Objective To investigate the effect of immunocompetent cells and immune regulator on the apoptosis of human mesenchymal stem cells ( MSCs) and on mRNA expression of heme oxygenase-1. Methods MSCs were cultured by density gradient centrifugation and then identified by flow cytometry. RT-PCR was used to detect HO-1 mRNA expression and flow cytometry was used to analyze cell apoptosis after the stimulation of IFN-? and PHA-activated T cells. Results The mRNA expression of Heme oxygenase-1 was observed in MSCs and decreased after the stimulation of IFN-? and activated T cells. IFN-?,znpp-Ⅸ and combined these two caused obvious cell apoptosis in MSCs,with an apoptotic rate of ( 56. 50 ? 0. 16) % ,( 56. 85 ? 2. 27) % ,and ( 82. 53 ? 2. 65) % respectively. All of them had a significant difference compared with the normal MSCs [( 7. 56 ? 1. 43) % ,P

Adult ; Diagnosis, Differential ; Female ; Humans ; Keratin-18 ; metabolism ; Mucin-1 ; metabolism ; Omentum ; metabolism ; pathology ; Peritoneal Neoplasms ; metabolism ; pathology ; ultrastructure ; Pregnancy ; Trophoblastic Neoplasms ; metabolism ; pathology ; ultrastructure ; Trophoblastic Tumor, Placental Site ; pathology ; Vimentin ; metabolism

OBJECTIVETo determine the physiological role of D-bifunctional protein (DBP) in bile acid biosynthesis through investigating the effect of increasing activity of DBP on bile acid biosynthesis.

METHODSTwenty male Wistar rats were divided into two groups: diethylhexyl phthalate (DEHP) group (n = 10) and control group (n = 10). Serum triglyceride, total cholesterol, hepatic DBP activity, and fecal bile acids were assayed. The mRNA levels of hepatic peroxisome proliferator-activated receptor alpha (PPARalpha), DBP, and cholesterol 7alpha-hydroxylase (CYP7A1) were detected by RT-PCR.

RESULTSCompared with control group, serum triglyceride level was decreased significantly and PPARalphamRNA level was increased significantly in DEHP group (P < 0.01). Together with a sharp induction of DBP mRNA expression and DBP activity in DEHP group (P < 0.01), the levels of CYP7A1 mRNA and fecal bile acids were significantly increased by 1.9 times and 1.6 times respectively compared to control group (P < 0.01). There was a significantly positive correlation between DBP mRNA level or DBP activity and CYP7A1 mRNA level (r = 0.89, P < 0.01; r = 0.95, P < 0.01).

CONCLUSIONThe up-regulation of DBP mRNA and activity in liver can result in the increase in CYP7A1 mRNA expression and bile acid biosynthesis, suggesting that DBP may be involved in bile acid biosynthesis together with CYP7A1.

17-Hydroxysteroid Dehydrogenases ; metabolism ; Animals ; Bile Acids and Salts ; biosynthesis ; Cholesterol 7-alpha-Hydroxylase ; analysis ; Enoyl-CoA Hydratase ; metabolism ; Liver ; metabolism ; Male ; Multienzyme Complexes ; metabolism ; PPAR alpha ; analysis ; Peroxisomal Multifunctional Protein-2 ; RNA, Messenger ; analysis ; Random Allocation ; Rats ; Rats, Wistar

In the present study, the specifically knockdown models of P-gp or MRP2 were constructed by using a series of chemically synthesized small interfering RNA (siRNA) in vitro. The expression of P-gp and MRP2 was measured by real-time PCR and Western blot, and the function was evaluated by applying P-gp and MRP2 substrate, rhodamine and methotrexate. The results showed that MRP2 siRNA-3 or P-gp siRNA-2 significantly decreased the mRNA expression of MRP2 or P-gp, the inhibition ratio was 68% or 84%; MRP2 siRNA-3 or P-gp siRNA-2 at a dose of 80 nmol x L(-1) significantly reduced the protein expression of MRP2 or P-gp at 48 h after treatment, the inhibition ratio was 62% or 70%. Meanwhile, other transporters were not influenced by siRNA. When pretreatment with MRP2 siRNA-3 or P-gp siRNA-2, the efflux of methotrexate or rhodamine decreased significantly and the intra-cellular concentration increased. The results suggested that chemically synthesized siRNA could significantly inhibit the expression and function of MRP2 and P-gp, and the model of RNAi in vitro could be used to evaluate the role of efflux transporters in transportation of drugs.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Gene Knockdown Techniques ; Multidrug Resistance-Associated Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction

BACKGROUND: Deep vein thrombosis (DVT) of the lower limbs is one of the severe complications of total hip replacement (THR)during the perioperative period. The incidence rate was high. The effect of primary disease on the DVT after THR in the elderly remains poorly understood.OBJECTIVE: To explore the effect of primary disease on DVT after THR in the elderly.METHODS: 147 cases with unilateral THR aged 64-93 years, were included and divided into two groups. Fracture group contained 68 cases, which of them were all traumatic femoral neck fracture ones. Osteopathia group contained 79 cases, and they had no traumatic injury. We selected total hip prostheses according to their physiological age, preoperative socialization ability,substantia ossea and life expectancy. Biological prosthesis in 5 cases and mixed prosthesis in 12 cases were used, while the others used bone cement prosthesis. If the patient had pain and/or swelling on the injured limb, with or without Homans/Neuhofs sign, we did the pressurized ultrasonic Doppler to examine whether the patient had got DVT.RESULTS AND CONCLUSION: In the fracture group, 32 cases had swelling in the injured limbs, and 20 cases with pain, 15 cases with Homans/Neuhofs sign, and 29 DVT cases were confirmed by the pressurized ultrasonic Doppler. One femoral neck fracture case of THR had no DVT clinical signs and was dead 17 days later. The autopsy found that it was an mixed type combined of pulmonary embolism; Osteopathia group: 20 cases had swelling in the injured limbs, and 11 cases of pain, 9 cases of Homans/Neuhofs sign, 20 DVT cases were confirmed by the pressurized ultrasonic Doppler. The blood coagulation state was greater, and the incidence rate of DVT in the lower limbs was greater in the fracture group compared with osteopathia group (P ＜ 0.05). These indicated that femoral neck fracture is a high risk factor for DVT development in lower limb affer THR in the elderly.

The cholesterol oxidase producing strain Brevibacterium sp.DGCDC-82 was treated with NTG (1 mg/mL)under ultrasonicztion(200 W,50 kHz).A red mutant named Brevibacterium sp.DGCCN-25 showed higher and stable production of cholesterol oxidase was obtained.The enzyme activity was increased by 140%,it is 1.24 U/mL.Then dealed with DGCCN-25 using the same method,two revertants were obtained,one was white and the other was rose pink.The enzyme activity of two revertants was obvious decrease,they are 0.17 U/mL and 0.69 U/mL.The results showed the positive correlation between COD acticity and red pigment producing by Brevibacterium sp..The relativity model can be used as a method of screening for mutation and directed evolution.

Objective To investigate the potential role of angiotensinⅡ(AngⅡ)-angiotensinⅡreceptor 1 (ATRI) pathway on inflammatory activation in the lung of rats. Method Twenty four Sprague-Dawley rats were randomly divided into four groups: control group, Ang II group, AngⅡ+losartan group and losartan group. Lung wet/dry weight (W/D) was recorded to assess lung injury. The total lung homogenates were prepared to detect nuclear factor-kappa B (NF-?B) activation by electrophoretic mobility gel shift assary (EMSA), tumor necrosis factor (TNF)-?mRNA expression by reverse transcription polymerase chain reaction (RT-PCR), myeloperoxidase (MPO) and malondialdehyde (MDA) by colorimetry. Plasma yon Willebrand Factor (vWF) were assessed by enzyme-linked immunosorbent assay (ELISA). Meanwhile, pathological changes were examined under optical microscope. Results Histologically, alveolar edema, hemorrhage, and massive inflammatory cell infiltration were observed in AngⅡgroup, but not in control group and losartan group. Compared with AngⅡgroup, histological injury was lesser in AngⅡ+ losartan group. In AngⅡgroup, lung W/D, NF-?B activation, TNF-?mRNA expression, MPO, MDA and vWF were markedly higher than those in the other three groups. There were not significant differences of lung W/D, NF-?B activation, TNF-?mRNA expression, MPO, MDA and vWF in control group, AngⅡ+ losartan group and losartan group. Conclusions Systemic infusion of AngⅡcould up- regulate inflammatory mediator expression and induce lung injury in rats. AngⅡ, acting mainly through ATRI, induced inflammatory activation in the lung of rats.

Objective To identify the diseases outcome of inpatients of prefectural hospital in Xinjiang,through analyzing the diseases outcome of inpatients of a prefectural hospital in Xinjiang from 2006 to 2008.Methods To analyze the diseases outcome of inpatients(20 533 cases)randomly selectedin hospital from 2006 to 2008 the date were aralgzed.according to year,gender,ethnicity,disease systems.Results The overall cured rate,improved rate,healed rate,fatality rate of inpatients was 42.10％(8663 cases),51.10％(10 463 cases),6.00％(1242 cases),0.80％(165 cases)respectively.The fatality rate of inpatients of above 60 years old,45 ～59 years old,15 ～44 years old is higher in inpatients of various age groups; The fatality rate of inpatients of tumor,circulatory system diseases,symptoms of menopause with the experimental and clinical body seen to be kind to the suffering of its is higher in inpatients of various systemic diseases,binary multinomial logistic regression shows that the disease outcome is influenced by gender,age,disease systems,hospital annual,four single factors,but not influenced by ethnicity.Conclusion The cured rate adding improved rate is over 90％ of common and frequently-occurring diseases in the prefectural hospital of Xinjiang province.The fatality rate is higher in inpatients who suffered from tumor or circulatory system diseases or symptoms of menopause with the experimental and clinical body seen to be kind to the suffering of its and the fatality rate of all age groups over 15 years old is higher.

Objective This study is about to detect the KCNJ5 gene variations in aldosterone-producing adenoma (APA) with primary hyperaldosteronism (PA),and to investigate the association of the KCNJ5 gene missense mutations with APA and PA.Methods A total of 46 APA tumors and their clinical characteristics were collected from Hypertension Center of the People's Hospital of Xinjiang Uygur Autonomous Region,and all the tumors were confirmed by pathology.All the samples of the coding region segments of KCNJ5 were detected by PCR and direct DNA sequencing to compare the different missense mutations in the tumor cells and peripheral blood cells and to analyze the association between the genotype and phenotype.Results Three missense mutations were found in 46 patients with APA:c.451G ＞ C/A (p.G151R) (5/46),c.503T ＞ G (p.L168R) (4/46),c.830T ＞ A (p.S209T) (12/46).S209T,as an unreported somatic mutation was observed.There were no missense mutations detected in the peripheral blood.Sex,age,systolic blood pressure,diastolic blood pressure,duration of hypertension,plasma potassium,urine potassium,aldosterone,plasma renin activity,aldosterone,and plasma renin activity ratio,as well as the rate of positive family history were compared between the mutants and wild-types.Systolic blood pressure and plasma potassium level among G151 R,L168R,and S209T had statistical differences.Systolic blood pressure in G151R was significantly higher than the other two groups,while the plasma potassium level was significantly lower than the other groups.Conclusions Three missense mutations were found in 46 patients with APA.The subjects with mutations had more serious condition than those without mutations after comparing their clinical phenotype.Besides,other different subtypes may exist in PA.

OBJECTIVETo observe the effects of panax quinquefolius saponin (PQS) of stem and leaf on glucose-lipid metabolism and insulin signal transduction in the insulin resistant model of adipocytes.

METHODSThe insulin resistant model of differentiated 3T3-L1 adipocytes was established in vitro with free fatty acid. After induction of insulin resistance, cells were treated with metformin or PQS for 2 days. The glucose consumption in culture fluid was detected by glucose oxidase method; the effects of PQS on the lipolysis induced by tumor necrosis factor (TNF-alpha) was observed using colorimetry; and the phospholation of signal proteins was detected by Western-blot.

RESULTSThe amount of glucose consumption (mmol/L) in the model group (5.250 +/- 2. 671) was significantly lower than that in the normal control group (14.133 +/- 1.305, P < 0.01), it increased in the meformin treated group (11.807 +/- 1.358), and the groups treated with high-, middle- and low-dose PQS dose-dependently (10.784 +/- 2.373, 10.217 +/- 1.237 and 9.984 +/- 2.006, respectively), significantly higher than that in the model group (P < 0.01). Upon TNF-alpha treatment, the concentration of free fatty acid (FFA) (nmol/ microg) in culture medium was 2.479 +/- 0.597, predominantly higher than that in the control group (1.320 +/- 0.538, P < 0.01), while it was 1.210 +/- 0.566 in the metformin group, 1.105 +/- 0.631 in high-dose PQS group, 1.108 +/- 0.260 in the middle-dose PQS group, 1.201 +/- 0.593 in the low-dose PQS group, all were lower than that in the TNF-alpha group (P < 0.05 or P < 0.01), and a dose-dependent tendency of PQS's action was seen. The tyrosine phosphorylation of insulin receptor and IRS-1 as well as Ser473 phosphorylation of PKB were lower in the model group than in the control group; they were insignificantly changed in the low-dose PQS group, but did show significant difference in comparing with those in the high-and middle-dose PQS groups or metformin group.

CONCLUSIONPQS can accelerate the glucose utilization and depress the lipolysis in adipocytes induced by TNF-alpha, which may be correlated with its promoting insulin signal transduction and improving insulin resistance in adipocytes.

3T3-L1 Cells ; Adipocytes ; drug effects ; metabolism ; Animals ; Glucose ; metabolism ; Insulin ; metabolism ; Insulin Resistance ; Lipid Metabolism ; drug effects ; Mice ; Saponins ; pharmacology ; Signal Transduction ; drug effects