This study was purposed to verify the binding part of human complement C3 to complement receptor III (CRIII) in monocytes, the peptide rC3B, including the binding-site, was expressed, purified and identified. rC3B, the binding part of human complement C3 to CRIII, was selected by computer-aided modeling and summarizing researches published. Then, rC3B gene fragment was amplified by PCR, and cloned into prokaryotic vector pQE30a. The fusion protein rC3B was expressed in E.coli M15 and purified by Ni(2+)-chelating affinity chromatography. The activity of rC3B was identified by Western blot and adherence assay with monocytes. The results showed that rC3B fragment was obtained, and a prokaryotic expression vector pQE30-rC3B was constructed. rC3B was efficiently expressed and purified. In Western blot, the target protein showed the activity of binding with C3 antibody, while the purified protein showed the activity of adherence with monocytes. It is concluded that the recombinant C3B was obtained and identified, and this study lay the basis for the further functional analysis of C3.
Cloning, Molecular ; Complement C3 ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Macrophage-1 Antigen ; genetics ; metabolism ; Receptors, Complement 3b ; biosynthesis ; genetics ; isolation & purification ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification

Adult ; Central Nervous System Diseases ; diagnosis ; surgery ; Hamartoma ; diagnosis ; surgery ; Humans ; Male

OBJECTIVETo evaluate the role of adiponectin in regulating tumor necrosis factor alpha (TNF alpha) production and preventing fulminant autoimmunological damage of hepatocytes following concanavalin A (Con A) injection into mice.

METHODSThree days after recombinant plasmids pAA-neo-mAd were injected into the mice via the tail veins, Con A was injected into the mice. Mice transfected with empty pAA-neo vector served as controls. The serum levels of alanine aminotransferase (ALT), TNF alpha and adiponectin were detected, and histological examination of livers was carried out at different time points after the Con A injection. All results were subjected to statistical analyses.

RESULTSHistological examinations showed that the damage in livers of mice with high serum adiponectin levels was milder than that of the controls. The serum levels of ALT and TNF alpha were both lower than those of the controls (P less than 0.01, respectively). Statistical analyses showed the serum levels of ALT was negatively related to the levels of adiponectin in the sera (r=-0.5034).

CONCLUSIONAdiponectin is effective in protecting hepatocytes from Con A-induced immunological injury. The mechanism of this protective effect may be caused by inhibiting the synthesis and/or release of TNF alpha.

Adiponectin ; blood ; pharmacology ; Alanine Transaminase ; blood ; Animals ; Concanavalin A ; adverse effects ; Female ; Immune System Diseases ; chemically induced ; pathology ; prevention & control ; Liver ; drug effects ; pathology ; Liver Diseases ; pathology ; prevention & control ; Mice ; Mice, Inbred BALB C ; Tumor Necrosis Factor-alpha ; blood

This study was aimed to establish a high-throughput luciferase reporter system, through which to screen and identify miRNAs directly targeting p21, and to explore the biological function and significance of these miRNAs. Molecular cloning technique was used to construct and identify two lentivirus-expressing vectors-pWPXL-Luc and pWPXL-Luc-P21-3'UTR, virus particles were collected after the pWPXL-Luc or pWPXL-Luc-P21-3'UTR vectors were co-transfected with the psPAX2 packaging plasmid and the envelope plasmid pDM2G into HEK-293T cells. Furthermore, two stable cell lines expressing luciferase singly or co-expressed luciferase and P21-3'UTR were established by transducing HEK-293 cells with recombinant lentivirus; the former was used as control in the following experiments. Finally luciferase activity of the latter stable cells was measured by using the luciferase reporter assay system. The results showed that high-titre recombinant lentivirus was produced and two stable cell lines were constructed, also to some certain, the luciferase activity was in direct proportion to the number of cells. In conclusion, the high-throughput luciferase reporter system is established successfully; using this system, the 28 miRNA that directly target P21 Cip1/Waf1 are screened experimentally.
Genes, Reporter ; Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus ; genetics ; Luciferases ; genetics ; MicroRNAs ; genetics ; Plasmids ; Transduction, Genetic

OBJECTIVETo review the outcome of repeated percutaneous sperm aspiration (PESA) and testicular sperm extraction (TESE) for intracytoplasmic sperm injection (ICSI).

METHODSForty-three cycles of 31 cases of azoospermic patients which underwent at least two PESA or TESE for ICSI from January 2001 to December 2002 were collected. The sperm retrieval, fertilization, implantation and clinical pregnancy were analyzed.

RESULTSTwenty-four cases underwent PESA and 7 cases underwent TESE. There were not any complications in these patients. Compared with the first cycle of 154 cases, the fertilization rate, implantation rate and clinical pregnancy rate were 78.39% vs 73.64%, 19.68% vs 18.38% and 34.88% vs 37.91%, respectively(P > 0.05).

CONCLUSIONSRepeated PESA or TESE is safe and well tolerated in azoospermic patients. Compared with the first cycle, the differences of repeated PESA or TESE cycles in fertilization rate, implantation rate and clinical pregnancy rate were not statistically significant.

Adult ; Azoospermia ; therapy ; Female ; Humans ; Male ; Middle Aged ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic ; methods ; Tissue and Organ Harvesting ; methods

OBJECTIVETo determine the effect of proton pump inhibitor (PPI) on in-stent restenosis (ISR) in patients receiving clopidogrel therapy.

METHODSTotal 439 patients underwent percutaneous coronary intervention (PCI) were enrolled in the study,including 250 post-PCI patients discharged on clopidogrel alone and 189 patients discharged on clopidogrel with PPI. The in-stent restenosis (ISR) ratio of the patients in these two groups were observed.

RESULTSDuring a mean follow-up period of (13 ± 5.9) months, the post-PCI patients discharged on concomitant clopidogrel-PPI therapy had higher risk of ISR than those discharged on clopidogrel alone (19.6% Compared with 8%, P<0.001).

CONCLUSIONConcomitant use of clopidogrel and PPI after hospital discharge would increase the risk of ISR for post-PCI patients.

Angioplasty, Balloon, Coronary ; Coronary Restenosis ; etiology ; Drug Antagonism ; Drug Therapy, Combination ; Follow-Up Studies ; Humans ; Platelet Aggregation Inhibitors ; therapeutic use ; Proton Pump Inhibitors ; therapeutic use ; Retrospective Studies ; Risk ; Stents ; Ticlopidine ; analogs & derivatives ; therapeutic use

OBJECTIVESTo review the retrospective treatment results of the azoospermia patients during January 2001 to January 2002 in the fertility center.

METHODSOne hundred males attempted intracytoplasmic sperm injection (ICSI) cycle for treatment of azoospermia. All patients were undergone sperm retrieval by percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE) while their wives received conventional ovarian hyperstimulation. The hormone levels, testicular histology, the rates of sperm retrieval, fertilization, implantation and pregnancy were analysed and evaluated.

RESULTSSperm were retrieved by PESA in 76 of 100 (76%) and by TESE in 23 of 100 (23%) men of azoospermia. The fertilization rate, implantation rate and clinical pregnancy rate were 71.3%, 20.35% and 42.11% respectively in PESA group, and 75.18%, 22.05% and 41.60% respectively in TESA group. Thirty-two clinical pregnancies were achieved with 15 ongoing pregnancies and subsequent live delivery for 15 cases in PESA group, and 2 cases of miscarriage, while 10 clinical pregnancies were achieved with 6 ongoing pregnancies, 2 cases of live delivery and 2 cases of miscarriage in TESA group. One case failed to retrieve sperm by TESE and canceled.

CONCLUSIONSHormonal levels and testicular histology are unable to predict which men with azoospermia will have sperm retrieved by PESA and TESE. PESA and TESE with ICSI are effective methods to treat azoospermia. There were no significant differences in fertilization, implantation and clinical pregnancy rate between two groups.

Adult ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Oligospermia ; blood ; therapy ; Pregnancy ; Pregnancy Rate ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; methods

Objective To analyze the therapeutic effects of microvaseular decompression on primary trigeminal neuralgia. Methods The general clinical data,culprit vessels and therapeutic effects of the surgical interventions were analyzed in 181 patients(aged from 24 to 79 years with a mean of54.9 years,including 78 male and 103 female patients)with established diagnosis of primary trigeminal neuralgia admitted from January,2000 to Deceber,2007. Results Blood type O was present in 43.65%of the 181 patients with trigeminal neuralgia,which had an increasing tendency compared to the national norm(33.8%).The morbidity ratio between the right and left side was 1.8:l in these patients.Forty-five patients(24.86%)were identified to have more than 2 culprit vessels.The culprit vessels included the superior cerebellar artery(96 cases),posterior inferior cerebellar artery(7 cases),anterior inferior cerebellar artery(25 cases),arteries communicated with veins(25 cases),internal auditory artery (13 cases),basilar artery(15 cases),vertical artery(9 cases),exclusive veins(15 cases,mainly vena pelrosa and bridging vein)and unknown vessels(9 cases).of the 181 cases,171(94.48%)were cured within one month,9(4.97%)showed relieved symptoms but required drug therapy,and 1 was in vegetative state(0.55%). Conclusions Patients with blood type O may have greater chance of developing primary trigeminal neuralgia.Microvascular decompression is an ideal treatment for primary trigeminal neuralgia,and clear identification of the culprit vessels can be crucial for decreasing the postoperative recurrence.

Objective To explore the treatments of complications in children after ventriculo-peritoneal shunt (VPS) and its complication appearance reasons.Methods The clinical data of 22 patients occurred complications after VPS,admitted to and performed VPS in our hospital from March 2005 to March 2011,were reviewed retrospectively.The reasons for the complications and treatment results were summarized.Results Obstruction of proximate tube appeared in 6 cases,obstruction of distal end in 4 cases,postoperative infection in 6 cases,subdural hydroma in 4 cases,and silt ventricle syndrome in 2 cases.The short-term response was 955％ after proper treatment.None died for related complications and one child with postoperative infection gave up treatment.Conclusion High risk of complications is noted in children performed VPS; active management based on the reasons of complications can acquire satisfactory curative effects.

BACKGROUNDHypertrophic obstructive cardiomyopathy (HOCM) carries an increased risk for sudden cardiac death. No data regarding the percutaneous transseptal myocardial ablation (PTSMA) and epicardial left ventricular pacing (LVP) were reported.

METHODSSeven patients with recurrent symptoms and increased resting left ventricular outflow tract pressure gradient (LVOTG) after PTSMA and another 14 patients with HOCM without history of PTSMA were studied. Both resting and dobutamine stress echocardiography, PTSMA and LVP were routinely performed.

RESULTSIn patients without previous PTSMA procedure, mild reduction of resting LVOTG was detected at 5 minutes after left ventricular pacing, and this reduction became significant at 10 minutes. All patients were divided into successful and unsuccessful groups according to their response to LVP. In contrary to patients in unsuccessful group, resting and R-S2 stimuli-induced LVOTG during PTSMA procedure were decreased dramatically ((9 +/- 5) mmHg vs (58 +/- 12) mmHg, (12 +/- 2) mmHg vs (113 +/- 27) mmHg, P < 0.001). Analysis of Logistic regression demonstrated that only LVOTG level during left ventricular pacing was an independent factor predicting the reduction of LVOTG immediately after PTSMA (odds ratio (OR), 0.59; 95% CI 2.67 to 5.82; P = 0.0002).

CONCLUSIONLeft ventricular endocardial temporary pacing plays a critical role in predicting acute effect on the reduction of LVOTG immediately after PTSMA procedure.

Adult ; Cardiac Pacing, Artificial ; Cardiomyopathy, Hypertrophic ; diagnostic imaging ; physiopathology ; therapy ; Catheter Ablation ; Echocardiography ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Pressure ; Ventricular Function, Left