The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.

Objective To study the clinical feature,treatment,and prognosis of the cytomegalovirus (CMV)pneumonia patients treated with immunosuppressor against kidney disease.Mlethod The patients received immunosuppressor against kidney disease in The First Affiliated Hospital of Sun Yat-sen University from June 1999 to December 2006.CMV antigen of leucocyte in the peripheral blood and/or bronchoalveolar lavage fluid of these patients were detected with immunocytochemical methods,and 21 patients were found suffering from CMV pneumonia.The 21 patients were introvenously injected with ganciclovir 5～10 mg/(kg?d),and the immunosuppressive agent treatment suspended.Their clinical feature and prognosis were retrospectively analyzed. Results The 21 patients received corticosteroids before CMV pneumonia contracted,of them,13 patients had been intensively treated with Methyllprednisolone with mean total dose(3.2?0.6)g.Of them,15 had been treated with cyclophosphamide with mean total dose(3.8?1.3)g.The median time from the beginning of using immunosuppressor to the onset of CMV pneumonia was 25(13～92)days.All patients had fever,cough, shortness of breath and X-ray showed interstitial pneumonia,of them,19 patients developed hypoxemia,and 11 patients' CMV antigen was positive in the leucocyte from bronchial lavage fluid.The result showed 9 patients survived and 12 died.The average duration of treatment with ganciclovir was(26.2?6.3)days. CMV pneumonia is a serious complication in patients who were treated with immunosuppressor against kidney disease.The mortality is high.Ganciclovir is a medicine of choice to treat CMV pneumonia.

Objective: To investigate the changes of myo cardial contractile function during myocardial stunning in calcium overload rats and the protective effects of tetrandrine. Methods: Forty-six rats were randomized into control, myocardial ischemia, myocardial stunning, low and high dose of tetrandrine groups. Another 10 rats were used to identify the calcium overload. vitamin D3 (0.3 million Unit/kg) and nicotinic acid were adm inistered. After 16 d when calcium overload occured, left anterior descending ar tery was ligated. Twenty minutes of myocardial ischemia followed by 60 min of re perfusion was induced. The contractile function parameters were determined dynam ically. At the end of experiment, myocardial cytosolic ［Ca2+］i was deter mined in various groups. In tetrandrine groups, tetrandrine (62.2 or 93.6 μmol/ kg ) was administered by gastrogavage daily.After 16 d, the rats undergone the e xperiments mentioned above. Results: Sixteen days after vitamin D3 , nicotinic acid were given, ［Ca2+］i increased by 2.6 folds (146.8±10.8 ) vs (368.5±22.6) nmol/L, (P<0.01). Whereas, ［Ca2+］i in tetrand rine groups were (210.8±16.4) and (198.6±15.3) nmol/L, which were significantl y lower than that of calcium overload group. Twenty minutes of myocardial ische mia resulted in the decrease of dp/dtmax and Vmax in all groups with the most si gnificant in stunning and calcium overload groups. The contractile function rest ored gradually after reperfusion. At all time points, dp/dtmax and Vmax in both tetrandrine groups were higher than those in both stunning and calcium overload groups. And effect with higher dose of tetrandrine were more significant than in low dose of tetrandrine. After 60 min of reperfusion, dp/dtmax in stunning, cal cium overload, low and high dose of tetrandrine groups were 49.7%, 51.5%, 71.0% and 83.4% of that in control, respectively， and Vmax were 55.0%, 49.8%, 73.9% and 77.5% of that in control, respectively. Conclusion: T he myocardial contractile function in vitamin D3-induced calcium overload gro up is impaired. On basis of myocardiocyte calcium overload, transient ischemia l eads to myocardial stunning. At the stage of ischemia, the impaired degree of my ocardial contractile function is similar to that in stunning group, suggesting a t this stage the effect of ischemia on myocardial function is greater than that of calcium overload. Tetrandrine chronically improves the myocardial function in Vitamin D3-induced calcium overload rats.

OBJECTIVETo investigate the expression of mammary serine proteinase inhibitor(Maspin) in patients with oral squamous cell carcinoma and its relationship with clinicopathological features.

METHODSThe Maspin protein expression was examined in 45 patients with oral squamous cell carcinoma and in non-malignant oral epithelia by immunohistochemical staining. The relationship between the Maspin protein expression and the clinicopathological parameters was statistically analyzed.

RESULTSThe Maspin protein expression in the cancerous tissues of oral squamous cell carcinoma was lower than that in the non-malignant oral epithelia (P = 0.001). Negative correlation was found between the Maspin protein expression in cancerous tissue and the status of lymph node metastasis (P = 0.038). Positive Maspin protein expression indicated negative lymph node metastasis, and negative correlation was also found between the Maspin protein expression and the postoperative metastasis (P = 0.004). Positive correlation was found between the Maspin expression and the patients' survival rate (P = 0.014), patients with positive Maspin protein expression having higher survival rate than those with negative Maspin expression.

CONCLUSIONSMaspin might be useful as a potential prognostic marker for oral squamous cell carcinoma.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Mouth Neoplasms ; metabolism ; pathology ; Serine Proteinase Inhibitors ; metabolism ; Serpins ; metabolism

OBJECTIVETo elucidate association between the mutation of nuclear factor of activated T cells 1 (NFATC1) gene in IPT-NFAT region and simple congenital heart disease (CHD) in children.

METHODWe used polymerase chain reaction (PCR) and the sequencing reaction to detect the mutations on the patients and their parents and (or) siblings.

RESULTSPCR amplification of the exon 7 region showed that 2 bands are obtained in 58% of patients with CHD and in 74% of their healthy parents and (or) siblings. Sequencing of the 2 bands revealed that both are amplicons of the exon 7 region, and that the additional band harbors an additional 44 nucleotides segment in the intronic region. The homozygous form of this allele was only present in patients with ventricular septal defect (2/24), atrial septal defect (3/18) and bicuspid aortic valve (1/4) in which G to A transition at nucleotide 17 of the third 44 bps was found. Neither the unrelated non-CHD individuals nor the ones with other CHD showed positive presence for the homozygous form of this allele.

CONCLUSIONSThere is a differential amplification of a tandem repeat region in intron 7 of NFATC1 and homozygous form of this allele in patients with ventricular septal defect, atrial septal defect and bicuspid aortic valve. NFATC1 gene may be an a susceptibility marker for ventricular septal defect, atrial septal defect and bicuspid aortic valve.

Adolescent ; Adult ; Aged ; Base Sequence ; Child ; Child, Preschool ; Female ; Genetic Testing ; Heart Defects, Congenital ; genetics ; Humans ; Infant ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation ; NFATC Transcription Factors ; genetics ; Pedigree ; Young Adult

BACKGROUNDThe aim of this study was to investigate DNA content and expression of c-erbB-2, PS2, and prostate-specific antigen (PSA) proteins in breast carcinomas with neuroendocrine (NE) cell differentiation.

METHODSChromogranin, c-erbB-2, PS2, and PSA in 131 samples of breast cancer were detected immunohistochemically. Classic Feulgen staining image analysis techniques were used to quantify DNA content in 81 of the breast cancer samples.

RESULTSThe c-erbB-2 positive rate in breast carcinoma samples containing neuroendocrine cells was 37.5% and the rate of high expression of c-erbB-2 (++ or +++) was 33.3%, both significantly lower than that in breast carcinomas without neuroendocrine cells (62.6% and 68.7%, respectively, P < 0.05). The rates of positive PS2 and PSA expression in breast carcinoma samples containing neuroendocrine cells were 72.2% and 55.0%, respectively, both significantly higher than that in breast carcinoma samples without neuroendocrine cells (45.0% and 16.4%, respectively, P < 0.05). In NE(+) samples, the integral optical density, DNA index, DNA stemline peak, > 5 c aneuploidy cells, and rate of aneuploidy among cells were all lower than that in NE(-) breast carcinomas (P < 0.01). In NE(+) grade I or II breast carcinomas, these indices were also all lower than that in the NE(-) breast carcinoma samples (P < 0.01).

CONCLUSIONBreast carcinomas with neuroendocrine differentiation have a lower rate of malignancy. Neuroendocrine differentiation could serve as a prognostic marker in clinical practice.

Breast Neoplasms ; chemistry ; pathology ; Cell Differentiation ; DNA ; analysis ; Female ; Humans ; Membrane Proteins ; analysis ; Neurosecretory Systems ; cytology ; Presenilin-2 ; Prostate-Specific Antigen ; analysis ; Receptor, ErbB-2 ; analysis

OBJECTIVETo explore the effects of manipulation and traction combined with Nimodipine on the blood flow velocity of vertebrobasilar artery (VBA) in cervical vertigo of high flow velocity,and to evaluate clinical therapeutic effects between two methods.

METHODSFrom March 2008 to Feburary 2009,70 patients who were diagnosed as high flow velocity of cervical vertigo were randomly divided into treatment group (35 cases) and control group (35 cases). Among 70 patients, 32 were male and 38 were female. The age ranged from 21 to 45 years with an average of 37.6 years. The disease course ranged from one day to two years with an average of 12.6 days. Patients of the treatment group were treated with manipulation for total three weeks, three times once week. The patients in the control group were treated with traction (weight ranged from 5 to 6 kg, 20 minutes each time, once every other day) and Nimodipine for total three weeks (three times each day, and with a dose of 40 mg each time). After three weeks, the changes of flow velacity of VBI and score before and after treatment were observed using transcranil Doppler (TCD) and Evaluation Scale for Cervical Vertigo. After six weeks, the therapeutic effects were assessed.

RESULTSThe mean velocity in left vertebral artery (LVA), right vertebral artery (RVA) and basilar artery (BA) were obviously lower than those before treatment in two groups (P < 0.01). The LVA, RVA and BA of the treatment group was lower than those of control group after 3 weeks (P < 0.01). There was significant difference in vertigo score after treatment between the two groups. The improvement rate of double-sides sign in X-ray image and the therapeutic effects of treatment group was superior to that of control group (P < 0.01).

CONCLUSIONThe effect of manipulation on flow velocity of VBA is superior to that of traction combined with Nimodipine, and there are better therapeutic effects in treating cervical vertigo of high flow velocity in comparison with traction combined with Nimodipine. But there are more higher demands for manipulation's application.

Adult ; Arteries ; physiopathology ; Blood Flow Velocity ; Cervical Vertebrae ; blood supply ; Female ; Humans ; Male ; Middle Aged ; Musculoskeletal Manipulations ; Vertigo ; physiopathology ; therapy ; Young Adult

OBJECTIVETo summarize the experience in the management of slow transit constipation complicated with adult megacolon.

METHODSThe clinical data of 32 above patients admitted between October 2007 and June 2011 were retrospectively studied.

RESULTSThirty-two patients were diagnosed as slow transit constipation according to the Roman III criteria. There were 15 males and 17 females aging from 18 to 56 years old. Sitz marker study showed prolonged colon transit time. Barium enema and defecography suggested bowel stricture locating in the transverse colon (n=3), descending colon (n=4), rectum (n=20), and concurrent transverse colon or descending colon and rectum (n=5). Anal manometry showed that anorectal inhibitory reflex was absent in 23 patients, while the other 9 patients were normal. Procedures performed included segmental colectomy and side-to-side anastomosis (n=1), subtotal colectomy and modified Duhamel anastomosis (n=16), total colectomy and ileal J-pouch Duhamel anastomosis (n=9). There were no postoperative complications. During the follow-up ranging from 3 to 47 months, defacatory function was excellent in 18, good in 9, and moderate in 5 patients.

CONCLUSIONSAdult megacolon should be considered differential diagnosis of slow transit constipation. Detailed history taking and thorough evaluation of testing is the key to obviate misdiagnosis. Extent of resection should include the diseased dilated colon and slow transit colon.

Adolescent ; Adult ; Aged ; Anastomosis, Surgical ; Constipation ; etiology ; surgery ; Digestive System Surgical Procedures ; Female ; Gastrointestinal Transit ; Humans ; Intestinal Obstruction ; Male ; Megacolon ; complications ; Postoperative Complications ; Retrospective Studies

OBJECTIVETo investigate the efficacy and safety of sirolimus in management of chronic allograft nephropathy (CAN).

METHODSA retrospective study was conducted involving 31 CAN patients followed up since March 2002, who experienced a change from a calcineurin inhibitor (CNI)-based regimen to a SRL-based regimen. Serum creatinine (Cr) in these patients was compared before and after the regimen change, and the adverse events associated with SRL were analyzed.

RESULTSTill March 2007 when the study closed, 15 patients reached the primary endpoint for resuming dialysis, 8 had improved and 8 had stable renal function. In patients with high Cr(0)(> or =3 mg/L, n=12), 9 resumed dialysis and 2 had improved renal function, but one of the patients with renal improvement eventually died due to infection; in the patients with low Cr(0)(<3 mg/L, n=19), 5 resumed dialysis, 8 had stable renal function and 6 had improved renal function, showing significant difference between the 2 groups (P=0.003). Altogether 14 patients reached the secondary endpoint for ceasing SRL for severe infection (5 patients, of whom 4 resumed dialysis and 1 died of infection) or adverse events associated with SRL (9 patients, of whom 4 resumed dialysis, 2 had stable and 3 had improved renal function). Hyperlipidemia (51.6%), leukocytopenia (41.9%), mouth ulcer (29.0%) and liver function lesion (16.1%) were the commonest adverse events in these patients, and totalling 13 severe adverse events were recorded, including 2 fatal cerebral hemorrhage, 3 fatal infection episodes, and 8 pulmonary and urinary infections that require hospitalization.

CONCLUSIONConversion from a CNI-based to SRL-based regimen can be effective for some CAN cases, especially for those with Cr(0) below 3 mg/L. Attention must be given to adverse events like hyperlipidemia and leukocytopenia, as well as the related cerebral vascular accidents and infections.

Adult ; Aged ; Chronic Disease ; Creatinine ; blood ; Female ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Kidney Function Tests ; Kidney Transplantation ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Sirolimus ; adverse effects ; therapeutic use ; Transplantation, Homologous ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate the efficacy and safety of oxymatrine in the treatment of chronic hepatitis B.

METHODSA multicenter randomized double-blind placebo-controlled trial was conducted. A total of 144 patients with chronic hepatitis B entered the study for 52 weeks; of them 72 received oxymatrine, and 72 received a placebo. Before and after the treatment, clinical symptoms, liver function, serum hepatitis B virus markers, and adverse drug reactions were observed.

RESULTSIn 144 patients, 14 were dropped and excluded due to inconsistencies in the included standard. Therefore, the efficacy and safety of 130 patients were analyzed. After being treated for 52 weeks, 70.77% of the patients in the study group had a normal ALT level, and in 43.08% and 33.33% their HBV DNA and HBeAg became negative. In the placebo group, 39.68% had normal ALT level, and 12.31% and 3.33% had their HBV DNA and HBeAg become negative. The rates of complete response and partial response in the oxymatrine group were 23.08% and 58.46%, and in the placebo group they were 3.08% and 44.62%. They were significantly higher in the oxymatrine group than in the placebo group. In the oxymatrine treated patients, 12 weeks after its withdrawal, 60.00% had a normal ALT level, 41.54% and 23.33% had both HBV DNA and HBeAg negative. In the placebo group, 31.75% had a normal ALT level, 3.08% and 1.67% had both HBV DNA and HBeAg negative. The rates of complete response and partial response in the oxymatrine group were 21.54% and 47.69%, and in the placebo group they were 0 and 41.54%. They were significantly higher in the study group than in the placebo group. The adverse reaction rates of oxymatrine in the study and the placebo group were 7.69% and 6.15%, respectively, but there was no statistical significant difference between them.

CONCLUSIONOxymatrine is an effective and safe agent for the treatment of chronic hepatitis B.

Adolescent ; Adult ; Aged ; Alkaloids ; adverse effects ; therapeutic use ; Antiviral Agents ; adverse effects ; therapeutic use ; Double-Blind Method ; Female ; Hepatitis B, Chronic ; drug therapy ; Humans ; Male ; Middle Aged ; Quinolizines