OBJECTIVETo determine the effect of type I transmembrane protein (IRE1alpha) deletions on the cell cycle of human periodontal ligament fibroblasts (hPDLFs) cells.

METHODSBased on the IRE1alpha deletions, a full-length model was successfully constructed. Moreover, overlapping polymerase chain reaction mutagenesis facilitated the establishment of two deletion mutants of IREla (pD-Kinase, pD-Rnase). The full-length model and two mutant eukaryotic expression vectors were transfected into hPDLFs cells. Western blot analysis was performed to identify the expression in the cells. The changes in the cell cycle of hPDLFS cells were detected by flow cytometry (FCM).

RESULTSThe two deletion mutants of IRE1alpha with eukaryotic expression vectors were successfully constructed and correctly expressed in hPDLFs cells based on Western blot analysis. Under stress conditions, the FCM assay showed that cell percentage of S phases increased, whereas that of G1 phases decreased in the IRE1alpha group (P < 0.05) compared with the control group of tunicamycin (TM) treatment. Moreover, the cell percentage of the S phases decreased, whereas that of the G1 phases increased in the D-Rnase group (P < 0.05) compared with the control. The deletion mutant D-Kinase had no influence on hPDLFS cell proliferation and cycle (P>0.05).

CONCLUSIONUnder stress conditions, IRE1alpha can improve the cell cycle of hPDLFs cells from the G1 to the S phase. The deletion mutant D-Rnase cause hPDLFs cell growth arrest at the G1 phase, whereas deletion mutant D-Kinase has no significant effect.

Cell Cycle ; Cell Proliferation ; Endoribonucleases ; Fibroblasts ; Humans ; Periodontal Ligament ; Protein-Serine-Threonine Kinases ; Transfection

Objective To explore the effects of diabetes mellitus on the left ventricular geometry in essential hypertension(EH). Methods Patients with hypertension admitted between 2000 to 2007 were retrospectively analysed.Those meeting the study criteria were divided into EH group(n=1660) and EH with diabetes mellitus group(n=953).The general data,biochemical parameters and echocardiographic findings were analysed. Results Compared with EH group,the age,course of hypertension,body mass index,systolic blood pressure,fasting blood glucose and triglyceride were increased(P

Objective To evaluate the effect of spleno-left adrenal vein shunt for the treatment of portal hypertensive upper GI bleeding caused by portal vein cavernous transformation in children.Methods Spleno-left adrenal vein shunt was performed in 8 children with portal hypertension due to cavernous transformation.The clinical data was reviewed.Results Portal vein pressure decreased significantly from (30±11)mm Hg to(22±7) mm Hg after shunt.There was no mortality perioperatively and during the follow-up.There were no recurrent hemorrhage nor hepatic encephalopathy occurring in the follow-up and all the children have normal intelligence and normal liver function though blood ammonia level increased significantly from(18±7)μmol/L to (60±17)μmol/L in 4 cases.In 7 cases in which preoperative whole blood cell count significantly decreased,the postoperative WBC,RBC,Hb and PLT was (7.64 ±4.46)×10~9/L,(4.54±0.97)×10~(12)/L,(133±5) g/L and (355.40±107.36)×10~9/L respectively (all P ＜0.05).In one case suffering from preoperative low PLT count the postop PLT reached 333×10~9/L,which was significantly higher than that preoperatively.Esophageal varices ameliorated in 6 cases.No stenosis of anastomotic stoma and thrombosis developed.Conclusion Spleno-left adrenal vein shunt is an effective procedure to treat portal vein cavernous transformation induced portal hypertension in children.

Objective To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) transplantation on angiogenesis and functional recovery after focal cerebral ischemia in rats.Methods BMSCs were isolated and cultured using the whole bone marrow adherent method,and conducted phenotypic identification using flow cytometry analysis of surface positive antigen of CD29,CD90 and the negative antigen of CD34,CD45.Rats were subjected to middle cerebral artery occlusion (MCAO) for 90 minutes,and divided into three groups randomly,the sham group,model group and BMSCs group.24 hours after cerebral ischemia,rats were injected with 1 ml BMSCs solution (1 × 106 cells/ml) or PBS via the tail vein.The modified neurological severity score(mNSS) test,the corner test and the adhesive tape test were used to evaluate sensorimotor function on the 1,7,14 and 28 days after ischemia.Infarcted volume was detected by toluidine blue staining,and the numbers of vWF positive microvessels and vascular endothelial growth factor (VEGF) positive cells in the ischemic boundary were determined by immunofluorescence.Results By flow cytometric analysis,the cell phenotype of passage 3 BMSCs showed that CD29,CD90,CD34 and CD45 were 98.3％,97.4％,0.2％ and 4.8％,respectively.Compared with the model group,BMSCs significantly reduced the score of mNSS(P＜0.01),the number of right turn of corner test(P＜0.05),latency of removal adhesive tape(P＜0.05) and the infarcted volume (P＜0.01).The numbers of vWF positive vesscls and the VEGF positive cells were (42.97±8.64)/mm2 and (54.83± 10.66)/mm2 at the boundary zone in model group 14 days after ischemia,respectively.BMSCs significantly increased the numbers of vWF positive vessels ((69.43± 7.29)/mm2) and VEGF positive cells ((78.70±6.16)/mm2,P＜0.01).Conclusion BMSCs can improve the functions of cerebral lesions after cerebral ischemia,which may be associated with the enhanced angiogenesis and VEGF expression in the ischemic boundary.

Objective To explore the effect of surgical treatment on primary gastrointestinal non-Hodgking lymphoma(NHL) in children.Methods Nine cases of clinical and follow-up data of primary gastrointestinal NHL were studied retrospectively to evaluate the effect of surgical treatment on primary gastrointestinal NHL in children.Results Seven cases were male and 2 cases were female.The mean age was(5.59?3.27)years old.The clinical manifestation included abdominal mass (7 cases),abdominal pain (5 cases),fever (2 cases),haematemesis and melena (2 cases),constipation (1 case) and paroxysmal abdominal pain with vomiting (1 case).Nine cases were diagnosed as primary gastrointestinal NHL,including 1 case of intussusception,1 case of acute appendicitis,2 cases of gastrointestinal obstruction,2 cases of gastrointestinal bleeding and 3 cases of abdominal mass.One case received the operation of intussusception reduction,tumor resection and intestinal anastomosis.One case received appendectomy.One case received the operation of tumor biopsy and transverse colon colostomy.Six cases received laparotomy.Six cases were diagnosed as Burkitt lymphoma.One case was anaplastic large cell lymphoma and 2 cases were diffuse large B-cell lymphoma.One case was at stage Ⅰ,1 case was at stage ⅠE,2 cases were at stage Ⅱ,3 cases were at stage ⅡE and 2 cases were at stage Ⅲ.Nine patients had received operation.One case died after operation and 8 cases had received combined chemotherapy.The 1 and 3 years survival rates were 75.0% and 37.5%,respectively.Conclusions Acute abdomen is often the first symptom of primary gastrointestinal NHL in children and comprehensive surgical treatment is an effective procedure for it.

The feasibility of simultaneously loading both liposoluble and water-soluble components of Salvia miltiorrhiza in emulsion was discussed, in order to provide new ideas in comprehensive application of effective components in S. miltiorrhiza in terms of technology of pharmaceutics. With tanshinone II (A) and salvianolic acid B as raw materials, soybean phospholipid and poloxamer 188 as emulsifiers, and glycerin as isoosmotic regulator, the central composite design-response surface method was employed to optimize the prescription. The coarse emulsion was prepared with the high-speed shearing method and then homogenized in the high pressure homogenizer. The biphasic drug-loading intravenous emulsion was prepared to investigate its pharmaceutical properties and stability. The prepared emulsion is orange-yellow, with the average diameter of 241 nm and Zeta potential of -35.3 mV. Specifically, the drug loading capacity of tanshinone II (A) and salvianolic acid B were 0.5 g x L(-1) and 1 g x L(-1), respectively, with a good stability among long-term retention samples. According to the results, the prepared emulsion could load liposoluble tanshinone II (A) and water-soluble salvianolic acid B simultaneously, which lays a pharmaceutical foundation for giving full play to the efficacy of S. miltiorrhiza.
Chemistry, Pharmaceutical ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Emulsions ; chemistry ; Quality Control ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo explore the effect of ligustrazine on the migration of bone marrow mesenchymal stem cells (BMSCs) and protein expressions of matrix metalloproteinase-2 and-9 (MMP-2 and MMP-9) in vitro.

METHODSBMSCs were in vitro isolated and cultured using whole bone marrow adherent method, and phenotypes [surface positive antigens (CD29 and CD90) and negative antigens (CD34 and CD45)] identified using flow cytometry. BMSCs were divided into the blank control group, 25, 50, 100 µmol/L ligustrazine group, and the GM6001 group (100 µmol/L ligustrazine +MMPs inhibitor GM6001 ). The migration of BMSCs was tested by Transwell chamber test and wound healing assay after treated with ligustrazine for 24 h. The protein expressions of MMP-2 and MMP-9 were detected by Western blot.

RESULTSThe third passage BMSCs grew well in uniform morphology. The expression rate of CD29, CD90, CD34, and CD45 was 96.9%, 97.3%, 0.2%, and 3.0%, respectively. Compared with the blank control group, the number of migrated cells and relative distance of cell invasion increased, and the protein expressions of MMP-2 and MMP-9 were elevated in each ligustrazine group (P < 0.05, P < 0.01). Compared with 100 µmol/L ligustrazine group, the number of migrated cells and relative distance of cell invasion decreased in 25 and 50 µmol/L ligustrazine groups and the GM6001 group (P < 0.01). Protein expression of MMP-2 decreased in 25 and 50 µmol/L ligustrazine groups (P < 0.01).

CONCLUSIONLigustrazine could promote the migration of BMSCs in vitro, and its mechanism might be related to up-regulating expression levels of MMP-2 and MMP-9 protein.

Cell Movement ; Cells, Cultured ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Pyrazines ; pharmacology ; Up-Regulation

Objective To present a preliminary latest procedure for portal hypertension and evaluate the technical feasibility and efficacy of portacaval shunt creation through the percutaneous transhepatic approach in order to make a hemodynamic comparison with that of the classic TIPS.Methods Thirty-eight patients with portal hypertension(36 men;mean age 57 years,range 32～73)were referred for PTPS procedure because of bleeding varices(n=36),intractable ascites(n=1),and hepatopulmonary syndrome(n=1).The severity of liver disease was classified as Child-Pugh B in 27 and C in 11.The PTPS was created by a percutaneous transhepatic puncture into right portal vein and then through left portal vein to the hepatic segment of IVC followed by a prefabrication stress stent-graft placement at the very site.Results Technical and functional success of 100% was achieved in all patients,without related complications.The postprocedural portal vein-IVC gradients mean 13 cmH_2O was achieved with the follow-up period mean 493 days.No recurrence of variceal bleeding and controlled refractory ascites were achieved,and still more with primary patency rate of the involved vascular structure up to 94.8% at 365 days,much better than classic TIPS. Conclusions Portacaval shunt creation using the prefabrication stress stent via percutaneous transhepatic technique is safe and feasible.The compact coincidence was obtained between the stent and the involved vessel with restoration of intrahepatic portal venous bemodynamics together with partial lowering of portal venous pressure and guaranteeing intrahepatic perfusion through right portal vein.It is also obviously to have postoperative prevention of shunt restenoses and lowering postoperative incidence of hepato-encephalopathy.

OBJECTIVETo study the epidemiology of human metapneumovirus (hMPV) infection in children and its relations with meteorological conditions in Suzhou.

METHODSamples obtained from 6655 children hospitalized with acute respiratory tract infections (ARIs) during the period from 2006 to 2009, were tested for virus pathogens. Nasopharyngeal aspirates were obtained from the children according to a standard protocol and were tested for respiratory syncytial virus (RSV), influenza viruses (IFV) A and B, parainfluenza virus (PIV) types 1, 2, and 3 and adenovirus (ADV) with direct immunofluorescence assay. Samples were tested for hMPV with reverse transcription polymerase chain reaction (RT-PCR). Meteorological conditions including mean temperature, relative humidity, rainfall amount, sum of sunshine and mean wind velocity were collected monthly. The relationship between activity of the virus and meteorological conditions was analyzed by linear regression and stepwise regression analysis.

RESULTViral pathogens were identified in 32.2% of 6655 specimens. The positive rate of hMPV was 8.9%, RSV was 15.7%, IFV, PIV and ADV detection rates were less than that of hMPV. The annual positive rate of hMPV from 2006 to 2009 was 8.2%, 8.1%, 12.7%, 7.4% respectively (χ(2) = 33.23, P < 0.05). The hMPV positive rate of the four seasons was 11.6%, 7.6%, 4.7% and 11.7%, respectively, detection rate in winter and spring was significantly higher than those in summer and autumn (χ(2) = 74.67, P < 0.001). The positive rate of hMPV and the monthly mean temperature was moderately correlated (r = -0.43), and the monthly average rainfall (r = -0.29), monthly mean relative humidity (r = -0.27), monthly average sunshine duration (r = -0.11), the monthly average wind speed (r = -0.13) had low correlations.

CONCLUSIONhMPV was the second most common viral pathogen of acute respiratory tract infection in children in Suzhou, which prevailed predominantly in the winter and spring. Climatic factors, especially temperature and rainfall may affect the prevalence of hMPV.

Child ; Child, Preschool ; China ; epidemiology ; Climate ; Female ; Humans ; Infant ; Male ; Metapneumovirus ; Respiratory Tract Infections ; epidemiology ; virology ; Seasons