Objective To investigate the effect of angiotenisn ⅡI (Ang Ⅱ) on RIN-m β-cell,and to explore the mechanism of β-cell function impairment caused by Ang Ⅱ.Methods RIN-m cells were cultured with various concentrations of AngⅡ (0.1,1,10,100 nmol/L).After incubation for 24 hours,the basal(3.3 mmol/L) and glucose-stimulated(16.7 mmol/L) insulin secretion(GSIS)were detected by radioimmunoassay,mRNA and protein expressions of uncoupling protein 2(UCP2)were determined by RT-PCR and Western blot,respectively.The intracellular ATP content was measured by luciferase bioluminescence.The mitochondrial membrane potential and cellular Ca~(2+) concentration were detected by flow cytometry.Results (1) Various concentrations of Ang Ⅱ had no significant influence on the basal insulin secrection of RIN-m cell(F=0.644,P = 0.634).Except for 0.1 nmol/L AngⅡ,the other concentrations of Ang Ⅱ markedly reduced GSIS of RIN-m cells(F= 118.528,P = 0.000).(2) Compared with the control group,Ang Ⅱ significantly increased mRNA and protein expression of UCP2(F= 1 370,P = 0.000;F=675.175,P = 0.000).(3)Except for 0.1 nmol/L Ang Ⅱ,the other concentrations of Ang Ⅱ significantly decreased the mitochondrial membrane potential,cellular ATP content,and cellular Ca2+ concentration of RIN-m cell(F=4.035,P=0.008;F=3.353,P = 0.013;F=5.867,P = 0.001).Conclusion Ang Ⅱ impairs GSIS of p-cell,the mechanism of impairment may be interpreted that Ang Ⅱ can increase the expression of UCP2,furthermore,it can reduce mitochondrial membrane potential,decrease the content of cellular ATP and the concentration of cellular Ca~(2+),can finally impair the function of β-cell.

Transplantation of the microencapsulated recombinant cells is a novel alternative approach to gene therapy of tumors. The semi-permeable membrane of microcapsule protects cells from host's immune rejection, increases the efficiency of gene transfer and reduces the need for frequent injection. Optimization of the preparation and culture is needed to acquire biological microcapsule with high cell viability and protein production. In this work, we studied the effect of different preparation and culture condition on the microencapsulated recombinant CHO cells growth and endostatin production. The result showed that the inoculum cells growth phase and seeding density potently affected the growth and endostatin production of the recombinant CHO cells in the microcapsule. The exponential growth phase recombinant CHO cells with a seeding density of 1 x 10(6) - 2 x 10(6) cells/ mL microcapsules benefited to the cells growth and endostatin production. The time of preparation was another important effect factor of cells viability, the cells viability decreased with the increase of preparation time and the time of preparation should be under 5h for maintaining the cell viability and endostain production. The highest viable cell density and endostatin production was acquired when the microcapsule percentage was 5% in the culture of the microencapsulated cells, the cell growth and endostatin production decreased with the increase of the microcapsule percentage.
Animals ; CHO Cells ; Capsules ; Cell Culture Techniques ; Cell Proliferation ; Cricetinae ; Cricetulus ; Endostatins ; metabolism ; Technology, Pharmaceutical ; instrumentation ; methods ; Time Factors

OBJECTIVETo isolate viruses from mosquitoes in the south of Xinjiang and identify these viruses primarily.

METHODSA total of 13 491 mosquitoes were collected in the south of Xinjiang from Jul to Aug, 2005. These mosquitoes were divided into 130 groups and grinded respectively. The supernates were inoculated in C6/36 and Vero cells. Viruses isolated were detected, the genomic nucleic types by electrophoresis of viral genomes and the morphologies observed under electronmicroscope.

RESULTSAll 42 viruses were isolated, which caused CPEs on C6/36 but not on Vero cells. 27 viruses showed similar genomic profiles with 12 dsRNA segments. 1 virus displayed genomic profile with 10 dsRNA segments. 5 viruses took on similar genomic profiles with about 4 kbp DNA band. 9 viruses did not get any taxonomy information. Electromicroscopic pictures of these viruses revealed that above four types of viruses had distinguished morphologies indicating different virus species.

CONCLUSIONThere should be several virus species in the mosquitoes in the south of Xinjiang. dsRNA virus with 12 genomic segments should play analysis a predominant role in the south of Xinjiang.

Animals ; Bluetongue virus ; classification ; genetics ; isolation & purification ; Cercopithecus aethiops ; China ; Culicidae ; virology ; Dengue Virus ; classification ; genetics ; isolation & purification ; Genome, Viral ; Insect Viruses ; classification ; genetics ; isolation & purification ; RNA, Double-Stranded ; genetics ; RNA, Viral ; genetics ; Reassortant Viruses ; genetics ; Sequence Analysis, DNA ; Vero Cells

BACKGROUNDInflammation within vulnerable coronary plaques may cause unstable angina by promoting rupture and erosion. C-reactive protein (CRP) is the most reliable and accessible test method for clinical use for identifying coronary artery disease event. Matrix metalloproteinase 9 (MMP-9) is highly over-expressed in the vulnerable regions of a plaque. Our aim was to evaluate the plasma levels of MMP-9 and hsCRP in subjects with both unstable angina and coronary plaques, as well as in those with unstable angina without coronary plaques.

METHODSPatients with newly diagnosed unstable angina pectoris from clinical presentation and ECG, who were undergoing coronary angiography from April 2007 to April 2009, were included in this study. A total of 170 subjects were enrolled in the study. Before angiography, the baseline clinical data (mainly including conventional risk factors) was collected. These patients were divided into two groups, a non-plaque group (G1) which included 55 patients with no significant stenosis or less than 20% stenosis in at least one of the major coronary artery branches, and a plaque group (G2) which included 115 patients with at least one of the major coronary artery branches unstable angina pectoris with at least 50% stenosis of one major coronary artery. The patients presenting with calcified nodules of a major coronary artery were excluded from this study. We examined the serum levels of MMP-9 for all cases by multi-effect enzyme-linked immunosorbent assay.

RESULTSThere was a significant difference in the serum levels of MMP-9 between the two groups (P < 0.001). The percentage of patients with hypertension, diabetes and current smokers were significantly different between the two groups (P = 0.034, P = 0.031, and P = 0.044 respectively). The univariate Logistic regression analyses of risk factors showed that smoking was the main risk factor for angina in the non-plaque group with the OR being 1.95 (95%CI 1.02 - 3.75). Hypertension, diabetes mellitus were negatively related with the occurrence of angina in the non-plaque group with the ORs being 0.50, and 0.36, respectively (95%CI 0.26 - 0.96 and 0.14 - 0.94). The MMP-9 level was negatively related to the occurrence of angina in the non-plaque group with an OR of 0.59 (95%CI 0.47 - 0.81).

CONCLUSIONSThere is a significantly difference in MMP-9 levels between the plaque and non-plaque groups. Current smoking has a significant influence on unstable angina patients without documented plaques. The serum MMP-9 level may be a significant biomarker which can help differentiate patients with unstable angina with plaques from those with unstable angina but without plaques.

Aged ; Angina, Unstable ; blood ; metabolism ; physiopathology ; C-Reactive Protein ; metabolism ; Coronary Angiography ; Coronary Artery Disease ; blood ; metabolism ; physiopathology ; Coronary Vessels ; metabolism ; pathology ; Female ; Humans ; Male ; Matrix Metalloproteinase 9 ; blood ; Middle Aged ; Multivariate Analysis ; Risk Factors ; Smoking ; adverse effects

OBJECTIVETo summarize the detailed failure mechanisms of revision hip arthroplasties and related risk factors.

METHODSFrom November 1988 to July 2008 revision of total hip arthroplasties was performed in 327 patients. The medical history, clinical and imaging material and operation records were investigated.

RESULTSRegarding revision as the end point of the study, the reasons for 327 revision arthroplasties were aseptic loosening in 226 hips (69.1%), infection in 52 hips (15.9%), periprosthetic fracture in 22 hips (6.7%), instability in 17 hips (5.2%), stem fracture in 5 hips (1.5%) and liner dissociation in 5 hips (1.5%).

CONCLUSIONSThe main failure mechanisms of primary hip arthroplasties are aseptic loosening and infection of implants, which could be attributed to improper selection of operation indications and implants and limitations to surgical philosophy and technique.

Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Periprosthetic Fractures ; Prosthesis Failure ; Reoperation ; Retrospective Studies ; Risk Factors ; Surgical Wound Infection ; Treatment Failure

OBJECTIVETo assess the clinical efficacy and the safety for periarthritis of shoulder treated with deep puncture by elongated needle combined with spike-hooked needle and cupping.

METHODSA multicentral randomized controlled study was adopted. One hundred and twenty cases were divided into an observation group and a control group, 60 cases in each one. In observation group, elongated needle, spike-hooked needle and cupping were used to stimulate Jianyu (LI 15), Naoshu (SI 10), Naohui (TE 13) and Jianqian (Extra). In control group, a conventional needling and cupping were applied to the same points as observation group. Visual Analogue Scale (VAS) and Melle evaluation for the function activity of shoulder joint were adopted to assess and compare the efficacy on analgesia and the effect on shoulder joint range of motion before and after treatment in two groups.

RESULTSIn observation group, the cured and markedly effective rate of analgesia was 91.7% (55/60), which was obviously better than 20.4% (12/59) in control group (P < 0.001). Concerning to the improvements of the range of motion in shoulder joint, the cured and markedly effective rate in observation group was 91.7% (55/60), which was better than 18. 6% (11/59) in control group (P < 0.001). VAS score and Melle score after treatment were apparently improved as compared with those before treatment in two groups (all P < 0.001). The improvements in observation group were superior to those in control group (both P < 0.001). There was no adverse event reported during the treatment in two groups.

CONCLUSIONThe deep puncture by elongated needle combined with spike-hooked needle and cupping achieves a significant clinical efficacy on periarthritis of shoulder and has no adverse reaction reported.

Acupuncture Therapy ; instrumentation ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Periarthritis ; physiopathology ; therapy ; Range of Motion, Articular ; Shoulder Joint ; physiopathology ; Shoulder Pain ; physiopathology ; therapy

OBJECTIVETo investigate the time trends of cancer mortality among residents in Kaifeng county, Henan province.

METHODSData on cancer mortality from the vital registration system in Kaifeng county from 1988 to 2005 was analyzed. A total of 9543 death records (5974 males and 3567 females) due to malignant tumors were studied. A two-year-period age-specified standardized mortality rates were directly adjusted by the world standard population, and the annual percentage change (APC) of mortality were estimated by a linear logarithm regression.

RESULTSThe crude cancer death rate for male was 95.09/100,000 and its age-standardized death rate was 117.41/100,000. While, the crude cancer death rate for female was 59.13/100,000 and the age-standardized death rate was 57.15/100,000. There was a significant growth tread for lung cancer (APC: 6.54%), liver cancer (5.07%) in males and breast cancer (7.04%) in females in the groups aged over 18. On the contrary, the decreasing treads for esophageal cancer in both of sexes (-7.09%, -13.53%) were also observed in this study. Meanwhile, there was no other significant changes in the trend, either in the tumor sites or mortality, was observed.

CONCLUSIONIn the past two decades, there has been a significant increasing trend for cancer mortality in Kaifeng county, of Henan Province. Hence, it is necessary to enhance epidemiological survey to identify risk factors at the earlier stages.

China ; epidemiology ; Death Certificates ; Female ; Humans ; Male ; Mortality ; trends ; Neoplasms ; mortality ; Rural Population

OBJECTIVETo obtain recombinant nestin and prepare anti-nestin polyclonal antibody (mAb) to explore the biological roles of nestin in the central nervous system development.

METHODSThe nestin cDNA was cloned from human neural stem cells by RT-PCR and ligated to prokaryotic expression plasmid pQE30 for construction of the recombinant vector pQE30-nestin. After sequencing, the recombinant vector was transformed into E.coli M15 and His-tagged nestin was induced by IPTG. The nestin was purified by Ni-NTA affinity chromatography column and characterized by SDS-PAGE and Western blotting. BALB/c mice were immunized with the purified recombinant protein to prepare the antiserum, which was analyzed by Western blotting, enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry.

RESULTSThe nestin gene was successfully cloned from human neural stem cells, which was identical to that reported in GenBank. After IPTG induction, the E.coli transformed with pQE30-nestin plasmid expressed a 25,000 His-tagged protein, which was successfully purified and identified as nestin by Western blotting. Western blotting, ELISA and immunohistochemistry demonstrated that the antiserum could specifically bind to the recombinant nestin as well as to nestin in fetal human and rat brains.

CONCLUSIONWe successfully cloned the nestin gene and expressed the nestin, and nestin mAb prepared can specifically recognize not only the recombinant nestin, but also nestin from human and rats brain tissues.

Adult Stem Cells ; cytology ; metabolism ; Animals ; Antibodies, Monoclonal ; immunology ; isolation & purification ; Blotting, Western ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Humans ; Immune Sera ; immunology ; Immunohistochemistry ; Intermediate Filament Proteins ; biosynthesis ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Nerve Tissue Proteins ; biosynthesis ; genetics ; immunology ; Nervous System ; cytology ; metabolism ; Nestin ; Recombinant Proteins ; biosynthesis ; immunology ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo evaluate the technique and clinical outcomes of modified transperitoneal laparoscopic radical prostatectomy.

METHODSA total of 105 patients received the operation with age ranging from 51 to 73 years from January 2008 to June 2010. Mean level of serum prostate specific antigen was 13.6 µg/L and mean prostatic volume was 45 ml. Pathological studies of biopsy confirmed the prostate carcinoma with Gleason score 6-8. Radionuclide bone scan revealed no metastasis. Based on previously retroperitoneal radical prostatectomy, modified technique was applied involving surgical approach, bladder neck dissection and vesicourethral anastomosis.

RESULTSMean operative time was 93 min (65 - 150 min). Intraoperative blood loss was 115 ml (50 - 400 ml). No complication of bowl injury occurred. Positive surgical margin was present in 24 patients. Normal continence were seen in 64 patients after catheter removed. Recovery of incontinence within 3 months was seen in 33 patients and 3 to 12 months in 5 patients respectively. Three patients with incontinence were still in the follow-up.

CONCLUSIONSTransperitoneal laparoscopic radical prostatectomy provides large working space and clear anatomic exposure. Higher efficiency and lower complication rate are obtained through modified laparoscopic technique involving seminal vesicle isolation, bladder neck dissection and vesicourethral anastomosis.

Abdominal Cavity ; surgery ; Aged ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Prostatectomy ; methods ; Prostatic Neoplasms ; surgery ; Retrospective Studies

OBJECTIVETo put some improvements to the traditional transforaminal lumbar interbody fusion (TLIF) and discuss its clinical significance.

METHODSCompleted the traditional posterior lumbar interbody fusion (PLIF) and TLIF procedure in 12 fresh cadavers, dissect further to expose the surrounding anatomical structures, and put the modified TLIF surgery according to the anatomical findings. And simulated the operation in 12 fresh cadavers, analyzed its feasibility and potential advantages.

RESULTSThe early anatomical study found that the related nerve root was in a state of high tension and certain risk of injury when completed the traditional PLIF and TLIF surgery, and found certain operational area between the superior articular process and the midline structures of the spinous processes and interspinous ligaments. Put the modified TLIF surgical approach according to the anatomical findings, which the operating area is located in PLIF outside and TLIF inside. As the following words: Take a posterior-middle incision, preserve the supraspinous and interspinous ligaments, and the spinous processes, dissect the bilateral paravertebral muscle, expose lamina and facet joints, not including transverse process, and remove unilateral inferior two third lamina, inferior articular process and expose the articular surface of the superior articular process, then dispose the intervertebral space for interbody fusion obliquely in the unilateral approach. Successfully completed the modified TLIF procedure in 12 fresh cadavers, the results showed that the technique has the following advantages. (1) Only remove unilateral inferior two third lamina and inferior articular process, preserve the supraspinous and interspinous ligaments, and the spinous processes, not expose the transverse process. (2) Both central canal, and lateral recess and nerve root canal of the operative side can be decompressed effectively simultaneously. (3) Avoid excessive traction to the thecal sac and traversing nerve roots and decrease the injury rate due to the reservation of the midline structures and the oblique manipulation, and less injury rate of the exiting nerve root, because of not necessary to expose it routinely.

CONCLUSIONSThe modified TLIF is safe and feasible, could effectively reduce the nerve roots injuries. Maybe it's a better choice for most of the Chinese patients at present.

Autopsy ; Humans ; Lumbar Vertebrae ; anatomy & histology ; surgery ; Spinal Fusion ; methods