Cryoglobulinemia ; physiopathology ; Humans ; Male ; Middle Aged ; Peripheral Nervous System Diseases ; diagnosis ; etiology ; Serologic Tests ; Syphilis ; diagnosis

Objective To establish a method of high resolution melting analysis (HRM) with unlabeled probe for detection of rt204 mutation in HBV P gene.Methods Plasmids with wild strain rt204M,mutant strains rt204I and rt204V were constructed,and the probes were designed and optimized.HRM plots were established by the constructed plasmids.A total of 185 samples were collected from patients with chronic hepatitis B in the Affiliated Hospital of School of Medicine of Ningbo University during May 2010 and May 2012.All samples were detected by HRM,and matched with characteristic melting pattern.rt204 mutations were screened,and then verified by DNA sequencing.Paired x2 test was used for the comparison of the detection of mutations.Results The melting temperatures for rt204I,rt204V and rt204M were 58.0℃,60.6℃ and 62.5℃,respectively.Among 185 samples,168 samples (90.8％) could be analyzed by HRM method,and 155 samples (83.8％) coule be successfully sequenced (P ＜0.01).In the 155 samples which were completely analyzed by HRM assay and sequencing,75 samples were rt204M,55samples were rt204I,and 25 samples were rt204V by using HRM method,with an overall mutation detection rate of 51.6％ (80/155) ; and by sequencing,110 samples were rt204M,30 samples were rt204I,and 15samples were rt204V,with an overall mutation detection rate of 29.0％ (45/155).The difference onmutation rates detected by the above two methods was of statistical significance (P ＜ 0.01).Conclusion HRM with unlabeled probe is simple,sensitive,rapid and specific for detection of rt204 mutation in HBV P gene.

Objective　To investigate the way of closed establishment of pneumoperitoneum(CEPP) in patients with peritoneal cvity adhesion in laparoscopy cholecystectomy(LC). Methods　CEPP experiences of 1046 patients in 6600 cases LC in our hospital from September 1991 to September 1999 were retrospectively analysed. The difficulty in establishing pneumoperitoneum was divided into two kinds: real establishment pneumoperitoneal difficulty(REPPD) and false establishment pneumoperitoneal difficulty(FEPPD). REPPD was due to Veress needle penetrating into visceral or extensive adhesion in peritoneal cavity resulting in CO2 flowing into difficulty. FEPPD was due to Veress needle pentrating in the fat out of peritoneum, in round hepatic ligament or in greater omentum. The formal situation needed to open laparotomy as a change, and the latter situation could establish pneumoperitoneum successfully by regulating the Veress needle penetrating direction or depth in the second penetration. Results　Of the 1046 patients, 1028 cases had been establishedpneumoperitoneum successfully though CEPP; 6 cases of REPPD and 18 of FEPPD were required opening laparotomy as a change. The successful rate of CEPP was 98.3%. Conclusions　CEPP is a safe and feasible method in patients with peritoneal adhesion in LC. It is the main reason for CEPP failure regarding REPPED as FEPPD made by the deficiency in LC experience and loss confidence in laparoscopist.

Objective To investigate the way of closed establishment of pneumoperitoneum(CEPP) in patients with peritoneal cvity adhesion in laparoscopy cholecystectomy(LC). Methods CEPP experiences of 1?046 patients in 6?600 cases LC in our hospital from September 1991 to September 1999 were retrospectively analysed. The difficulty in establishing pneumoperitoneum was divided into two kinds: real establishment pneumoperitoneal difficulty(REPPD) and false establishment pneumoperitoneal difficulty(FEPPD). REPPD was due to Veress needle penetrating into visceral or extensive adhesion in peritoneal cavity resulting in CO 2 flowing into difficulty. FEPPD was due to Veress needle pentrating in the fat out of peritoneum, in round hepatic ligament or in greater omentum. The formal situation needed to open laparotomy as a change, and the latter situation could establish pneumoperitoneum successfully by regulating the Veress needle penetrating direction or depth in the second penetration. Results Of the 1?046 patients, 1?028 cases had been establishedpneumoperitoneum successfully though CEPP; 6 cases of REPPD and 18 of FEPPD were required opening laparotomy as a change. The successful rate of CEPP was 98.3%. Conclusions CEPP is a safe and feasible method in patients with peritoneal adhesion in LC. It is the main reason for CEPP failure regarding REPPED as FEPPD made by the deficiency in LC experience and loss confidence in laparoscopist.

Objective To observe the effect of healthy education on BMD and the risk of bone fracture in pa- tients with primary osteoporosis.Methods 628 patients with primary osteoporosis were randomly divided into two groups under the same conditions,and healthy education were conducted in treated group but were not conducted in control group.Two years later,BMD and the rates of bone fractures were collected and analysed.Results Compared with control groups,the bone loss was significantly lower at the feroral neck,Ward's triangle and the great trochanter,and the risk of bone fractures was remarkably reduced in treated group(P

Objective To study the role of urinary kidney injury molecule-1 (KIM-1) in pediatric Henoch-Sch?nlein purpura (HSP). Methods Urinary levels of KIM-1 were examined using ELISA in 48 children with HSP including 23 HSPN children (HSPN group) and 25 non-HSPN children (HSP group), and 20 healthy children. The levels of urinary creatinine and 24-hour urine protein were also detected. The results were analyzed and compared among groups. Results The ratio of urinary KIM-1/creatinine (Cr) in HSPN children was signiifcantly higher than that in the other two groups (P<0.05). There was no signiifcant difference in the ratio of urinary KIM-1/Cr between HSP group and the control group (P>0.05). The ratio of urinary KIM-1/Cr had no correlation with 24-hour urine protein in all HSP children (r=0.239, P=0.590). Conclusions Urinary KIM-1 may play a role in the pathogenesis of pediatric HSPN.

Objective To study the different ultrasonic features in patients of polycystic ovary syndrome (PCOS) with or without obesity based on body mass index (BMI), and to investigate whether certain hormonal factors correlate with ovarian morphology and blood flow, and to discuss the role of ultrasound combined with hormone test in the diagnosis of obese PCOS. Methods One hundred and five women with PCOS were recruited. Patients were divided into two groups according to BMI;obese PCOS group (OB-PCOS, n=32, BMI≥25 kg/m2) and non-obese PCOS (NOB-PCOS, n=73, BMI<25 kg/m2). The ultrasonic parameters of follicle number (FN), ovarian volume (Vol), resistance index (RI) of ovarian stromal blood, RI of uterine artery and serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), the ratio of luteinizing hormone and follicle-stimulating hormone (LH/FSH), progesterone (P), estradiol (E2), free testosterone (FT), prolactin (PRL), sex hormoe binding globulin (SHBG), fasting plasma glucose (FPG), fasting insulin (FINS), the extent of insulin resistance and hyperandrogenism (HOMA-IR) were measured and compared. The correlation of the ultrasonic parameters and hormonal factors were analyzed. Results The Vol of OB-PCOS group was significantly higher than NOB-PCOS group [(12.25±4.89) ml vs (10.73±2.30) ml, t=2.20, P < 0.05]. FN and uterine artery RI of OB-PCOS group had a rising trend and RI of ovarian interstitial was on a reducing trend compared with NOB-PCOS group. But the differences were not statistically significant. The levels of FINS and HOMA-IR in OB-PCOS group [(14.82±6.45) mU/L and (3.91±3.30)] were significantly higher than those in NOB-PCOS group [(8.04±4.57) mU/L and (1.64±1.20)] (t=4.87, 3.47, respectively, both P < 0.01). And FSH in NOB-PCOS group was significantly higher than OB-PCOS group [(5.95±1.91) U/L vs (4.65±1.88) U/L, t=-2.77, P<0.01]. In POCS patients, FN was significantly associated with LH/FSH (r=0.35, P<0.01), and FT (r=0.38, P<0.01). Vol was significantly associated with LH/FSH, BMI, HOMA-IR and FPG (r=0.27, P<0.05;r=0.25, P<0.05;r=0.40, P<0.01;r=0.32, P<0.01). RI of ovarian stromal blood flow was significantly associated with SHBG (r=0.28, P<0.05). In OB-POCS group, RI of uterine artery was significantly associated with PRL (r=-0.58, P < 0.05). Vol was significantly associated with HOMA-IR (r=0.47, P < 0.05). In NOB-POCS group, FN was significantly associated with LH/FSH (r=0.33, P<0.05), and FT (r=0. 56, P<0.05). Vol was significantly associated with FT (r=0.31, P < 0.05). Conclusion There are some differences in the ultrasound and endocrine parameters between obese and non-obese PCOS patients, and some correlations exist between them.

Objective To investigate the correlation between acquired drug resistance-related genes and mobile genetic elements from pandrug-resistant Acinetobacter baumannii. Methods Fifty-three horizontal transfer drug resistance-related genes ( β-lactamases,aminoglycoside and quinolones resistance related) and 12 mobile genetic elements (including zygosity plasmid,transposon,insertion sequence and integron) were detected by polymerase chain reaction (PCR) in 20 clinical isolates of pandrug-resistant Acinetobacter baumannii.Index cluster analysis was performed to explore the correlation.Results In 20 strains of pandrug-resistant Acinetobacter baumannii,there were 3 types of β-lactamases related genes (TEM-1,ADC-30,OXA-23 ),4 types of aminoglycoside modifying enzyme genes [ aac (3)-Ⅰ,aac(6′)-Ⅰ b,ant( 3″)-Ⅰ and aph( 3′)-Ⅰ ],and 5 kinds of mobile genetic elements ( int Ⅰ 1,tnpU,tnp513,IS26 and ISAba1 ). Index cluster analysis showed high correlations between resistance genes [TEM-1,ADC-30,aac( 6′)-Ⅰ b,ant( 3″)-Ⅰ,abeB,qacE Δ1] and mobile genetic elements ( int Ⅰ 1,tnpU,tnp513,IS26,ISAba1 ).Conclusion Clinical isolated pandrug-resistant Acinetobacter baumannii carries several acquired drug resistance-related genes and mobile genetic elements,and there may be a close association between them.

Objective To explore the prevalence of 11 efflux pumps in isolates of multidrugresistant Escherichia coli(MDR-ECO). Methods Efflux pumps emrB, emrD , emrE, mdfA, sugE, mdtl,tehA, oqxA, qacE△1, qacE and smr-2 were detected by polymerase chain reaction(PCR)in 20 MDR-ECOs isolated from clinical samples. Results Efflux pumps emrB, emrD, emrE, mdfA, sugE, mdtI, qacE △1, tehA and oqxA were detected, and 8 efflux pumps were found in the same strain. Conclusion Multidrug- resistance in Escherichia coli may be related with efflux pumps.

BACKGROUND: Antigene RNAs (agRNAs) could be a useful tool to downregulate beta arrestin 2 (Arrb2) gene expressions, and realize gene knock-out effect in cell levels. OBJECTIVE: To observe the effects of agRNAs on DAMGO-induced mu-opioid receptor (MOR) desensitization in C17.2 cells by using agRNAs complementary to transcription start sites of beta arrestin 2 (Arrb2) to downregulate the gene expression in mouse C17.2 cells. METHODS: Mouse neural stem cells C17.2 was cultured in Dulbecco's minimal essential medium containing 10% fetal bovine serum, 10 mg/L penicillin and 10 mg/L ampicillin with 5% CO_2 at 37 ℃. The cells were passaged every 5 to 6 days after digestion with 0.25% trypsin when cells were 80% confluent. The expression of MOR on mouse C17.2 cells was detected by RT-PCR and immunocytochemical method. AgRNAs which could silence the expression of Arrb2 was transfected into C17.2 cells with Lipofectamine. The expression of green fluorescent protein gene was observed by fluorescence microscopy 24 hours after transfection. [~(35)S]GTPγS binding was assessed by autoradiography to examine the ability of the MOR to couple to G proteins on stimulation with selective agonist DAMGO.RESULTS AND CONCLUSION: The expression of MOR on C17.2 cells was confirmed by RT-PCR. The receptors were expressed on cell membrane and in plasma determined by immnocytochemistry. The expression of green fluorescent protein gene could be observed in C17.2 cells transfected with plasmid pGPU6/GFP/Arrb9 using fluorescent microsocpe. The results of [~(35)S]GTPγS binding showed that the stimulation ratio in cells with and without DAMGO stimulation or transfected with agRNAs were (113±14)%, (253±17)% and (239±15)% respectively. This indicated that agRNAs could downregulate the expression of beta-arrestin 2 and inhibit the desensitization of MOR.