OBJECTIVE To design the detection system of the leaky surface acoustic wave(LSAW) biosensor in(liquid) phase and detect human papilloma virus(HPV) target sequence with the LSAW biosensor.METHODS A detection delay line and a reference delay line were constructed,the phase of the two delay line changes was(evaluated) in liquid phase,and real time monitoring of HPV reaction was performed by using the LSAW biosensor system.RESULTS The phase of the two delay lines had different changes in liquid phase;HPV could lead to a great change of the phase in the detection delay line,and reference delay line had no change.CONCLUSIONS The(detection) system of the LSAW biosensor in liquid phase is successfully constructed and real time monitoring of(detection) of HPV is realized.

Objective To assess the efficacy of mycophenolate mofetil (MMF) combined with prednisone in the treatment of idiopathic membranous nephropathy (IMN) patients with positive serum phospholipase A2 receptor (PLA2R) antibody.Methods An open prospective study was performed on twenty-four biopsy-proven IMN patients with positive serum PLA2R antibody in Guangzhou First People''s Hospital from June 2012 to June 2016.The 24 patients were divided into two groups: MMF group in which MMF combined with prednisone was given for 12 months and CTX group in which intravenous cyclophosphamide (CTX) was monthly given combined with oral prednisone.Results After 6 months of immunosuppressive therapy,complete remission and partial remission rates were 25.0% vs.16.7% and 25.0% vs.25.0% in the MMF group and CTX group,respectively (P>0.05).In the MMF group and CTX group,serum PLA2R antibody in the same amount (8/12,66.7%) of patients turned negative.At the end of twelve-month treatment,all patients with negative PLA2R antibodies achieved complete or partial remission.Clinical remission (including complete and partial remission) rates in the MMF group and CTX group were both 66.7%.After immunosuppressive therapy,the levels of proteinuria and serum albumin in the two groups were significantly improved,but no significant difference were found between the two groups (proteinuria:F within-grouP=98.688,P<0.01;F between-grouP=0.133,P=0.719;F cross-grouP=1.223,P=0.304;serum albumin:F within-grouP=30.629,P<0.01;F between-grouP=0.137,P=0.715;F cros-grouP=0.455,P=0.565).At the end of six and twelve months of treatment,the proteinuria (after six months,MMF group: (2 893±2 515) mg/g vs.(6 236±2 117) mg/g,t=-3.522,P=0.002;CTX group: (2 690±2 254) mg/g vs.(5 386±2 447) mg/g,t=-2.808,P=0.010;after twelve months,MMF group:1 025(99-4 635) mg/g vs.(6 236±2 117) mg/g,Z=-3.291,P<0.0005;CTX group: (775(41-3 517) mg/g vs.(5 386±2 447) mg/g,Z=-3.118,P=0.001) and serum albumin levels (after six months,MMF group: (28.5±9.7) g/L vs.(19.8±4.4) g/L,t=2.841,P=0.012;CTX group: (29.0±7.6) g/L vs.(22.3±4.1) g/L,t=2.690,P=0.016;at the end of twelve months of treatment,,MMF group: (32.4±8.5) g/L vs.(19.8±4.4) g/L,t=4.570,P<0.0005;TX group: (32.2±7.9) g/L vs.(22.3±4.1) g/L,t=3.862,P=0.001) of the two groups were better than those prior to treatment.Conclusion For the IMN patients with positive serum PLA2R antibody,MMF combined with prednisone was as effective as conventional CTX combined with prednisone in the negative conversion of PLA2R antibody and the remission.The negative conversion of PLA2R antibody after 6 months of immunosuppressive treatment was an important indicator of predicting the remission.

Objective To study the preventive effects of vitamin K2 on tumor recurrence in patients with hepatocellalar carcinoma (HCC) after radical resection. Methods The clinical data of 50 patients with HCC who received radical resection from March 2006 to March 2007 in No. 181 Hospital of PLA were analyzed retrospec-tively. All the patients were divided into 2 groups according to the random number table. Twenty-six patients in vitamin K2 group were administered with menatetrenone (45 mg per day), and the rest 24 pateints were in the control group. The accumulative and tumor-free survival rates, differences between the 2 groups, multivariate factors for prognosis were analyzed by Kaplan-Meier curve, Log-rank test and Cox regression model, respectively. Results During a period of 36 month follow-up, 10 patients died and 28 had tunor recurrence. The 1-, 2-, 3-year accumulative survival rates were 96%, 92% and 83% in vitamin K2 group, and 96%, 82% and 63% in control group (χ2 = 3.61, P > 0.05). The 1-, 2-, 3-year tumor-free survival rates were 92%, 60% and 38% in vitamin K2 group, and 75%, 42% and 12% in control group, with significant difference between the 2 groups (χ2 =5.61, P <0.05). Univariate and multivariate Cox proportional hazard analysis showed that without taking menate-trenone, the preoperative level of alpha fetoprotein (AFP) ≥800 μg/L and vascular invasion were the indepen-dent risk factors for tumor recurrence. Conclusions Vitamin K2 has a suppressive effect on tumor recurrence of HCC, while patients with AFP≥800 μg/L before operation or with vascular invasion have poor prognosis.

Objective To observe the effects of psychological supportive therapy on the activities of daily living (ADL) and depression after cerebral hemorrhage. Methods 60 cases with mild or medium depression according to 17 items Hamilton Rating Scale for Depression (HAMD-17) after cerebral hemorrhage were divided into treatment group (n=30) and control group (n=30). All of them accepted physical exercise, and those in the treatment group accepted psychological supportive therapy in addition. They were assessed with Functional Independence Measure (FIM) and HAMD-17 before and after the treatment. Results After 2 months of treatment, the score of FIM increased more in the treatment group than in the control group (P<0.001), and the score of HAMD-17 decreased (P<0.001). Conclusion Psychological supportive therapy may promote the recovery of ADL and depression for patients post cerebral hemorrhage.

Objective To explore the effectiveness of a respiratory function training instrument with stable chronic obstructive pulmonary disease (COPD) patients.Methods Sixty-seven COPD patients in the stable period were randomly divided into a treatment group of 36 and a control group of 31 using a random number table.Both groups were given conventional pulmonary rehabilitation,including half-closed lip respiration,abdominal respiration and upper limb training.The treatment group was additionally provided with 30 minutes of respiratory training using a respiration function training instrument 5 times per week for 6 months.Both groups were assessed for their mobility,life quality and pulmonary function using the 6-minute walk test (6 MWT),a COPD assessment test (CAT),the BODE index,forced vital capacity (FVC),forced expiratory volume in one second (FEV1) and surface electromyography (SEMG) of the respiratory muscles before and after the 6-month intervention.Results Before the treatment there were no significant differences between the two groups in terms of any of the measurements.After the treatment,significant improvement was observed in the average 6 MWT,CAT,BODE index and SEMG results in both groups,but with significantly greater improvement in the treatment group.The average FVC and FEV1 results did not improve significantly,so after the intervention there was still no significant difference between the groups.Conclusions Respiratory training using the pulmonary function training instrument can improve the mobility,life quality and the functioning of the respiratory muscles of COPD patients in the stable period.

ObjectiveTo investigate the relationship between metastasis-associated gene 1 ( MTA1 )expression and invasive and metastatic ability of cervical cancer cell. MethodsThree kinds of plasmids pcDNA3( control group), pcDNA3-MTA1 ( MTA1 group) and pSilencer3. 1-MTA1-siRNA ( MTA1-siRNAgroup) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability. ResultsCompared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P＜0. 05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ±6, 169 ± 10 and 57 ±5,respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group(P ＜ 0. 05 ). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69. 3 ± 3. 6) ％, ( 80. 4 ± 5. 6 ) ％ and ( 39. 2 ± 7.4 ) ％ separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P ＜0. 05 ). In the migration assay, the number of cells migrated to the bottom side of the membrane in control,MTA1 and MTA1-siRNA group were 153 ± 17,247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19,354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA 1-siRNA group showed lower cell migration and invasion ability (P ＜ 0. 05 ). In cell cycle experiment, no significant differences of cell proportions including G1, S and G2 stage were found among three groups (P ＞ 0.05).In animal experiment, compared with control group,MTA1 group showed accelersted tumor formation and growth,whilethe MTA1-siRNA group showed the reverse effect ( P ＜ 0. 05 ). ConclusionsMTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.

OBJECTIVETo evaluate the efficacy and safety of Yigan Fupi Decoction (YFD) in the treatment of irritable bowel syndrome with diarrhea (IBS-D) patients.

METHODSA randomized controlled clinical trail was carried out in patients with IBS-D. All patients were randomly assigned to the treatment group (58 cases, treated with YFD) and the control group (58 cases, treated with Pinaverium Bromide Tablet). The treatment course was 4 weeks for all patients. The total effective rate, the stool property and state, the quality of life (QOL), and TCM syndrome efficacy were assessed by IBS bowel symptom severity scale (IBS-BSS), IBS defecation state questionnaire (IBS-DSQ), IBS quality of life questionnaire (IBS-QOL), and traditional Chinese medicine pattern curative effect scoring system (TCM-PES) before and after treatment.

RESULTSThere was no statistical difference in the total effective rate between the two groups (82.76% vs. 77.59%, P > 0.05). The treatment group was superior in the total IBS-BSS integral to the control group (P < 0.05). The total effective rate of improving the stool property was better in the treatment group than in the control group (81.03% vs. 72.41%, P < 0.05). Besides, the number of days for emergent defecation among 10 days was less in the treatment group than in the control group (P < 0.05). The improvement of the total IBS-QOL integral and the total integral of TCM syndrome were better in the treatment group than in the control group (P < 0.01). The total effective rate of TCM-PES was better in the treatment group than in the control group (84.48% vs. 70.69%, P < 0.05).

CONCLUSIONYFD was effective in the treatment of IBS-D patients of Gan-qi invading Pi syndrome, and could effectively relieve bowel symptoms, improve the stool property and the defecation frequency, elevate their QOL, and attenuate Gan-qi invading Pi syndrome with favorable safety and compliance.

Adult ; Diarrhea ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Irritable Bowel Syndrome ; drug therapy ; Male ; Middle Aged ; Morpholines ; therapeutic use ; Quality of Life ; Treatment Outcome

Objective To investigate mechanisms underlying the prevention of experimental autoimmune encephalomyelitis (EAE) in mice by intraperitoneal infusion of myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) (MOG i.p.).Methods C57BL/6 mice were immunized with MOG35-55 to establish the model of EAE and then were intraperitoneally injected daily with MOG35-55 or ovalbumin (OVA, served as control) from day 6 to day 16.EAE was evaluated daily using a general clinical scoring system and histological analysis.Numbers of lymphocytes in peripheral blood and central nervous system (CNS) were detected at different time points.Effects of MOG i.p.on the migration of MOG-T cells in vivo were analyzed by an adoptive transfer experiment.Maturation of splenic antigen-presenting cells (APCs) and migration of MOG-T cells in vitro were examined by fluorescence activated cell sorting (FACS) and a Transwell system, respectively.Results MOG i.p.protected the mice from development of EAE by blocking the lymphocyte recruitment to CNS.More effector T cells were trapped in the periphery of EAE and naive mice in adoptive transfer experiment after MOG i.p.treatment.MOG i.p.induced the maturation of splenic APCs and enhanced the expression of CD80, CD86 and major histocompatibility complex class Ⅱ (MHCⅡ) molecules.Mature APCs blocked the recruitment of effector T cells to CNS.Conclusion MOG i.p.protects mice from EAE by inducing the maturation of splenic APCs.

The Coat protein and Maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32aCP.The conservative sequence of FMDV internal ribosome entry site(IRES) was cloned into the downstream of pET32aCP bacteriophage gene to construct the prokaryotic expression vector pCPES.The recombinant plasmid pCPES transformed into E.coli strain BL21(DE3) was induced to express with 1mmol/L IPTG.The expression products were purified by sucrose density gradient centrifugation.The expression products observed by TEM were circular viruslike particles,and the diameter of these particles was about 26nm.The stability of viruslike particles was detected,and the viruslike particles was identified by RTPCR.The results showed that the viruslike particles contain the FMDV IRES RNA and have good stability.The viruslike particles have great prospect as the standard and quality control in the area of RNA virus detection.

Objective: To study the factors effecting the expression of the reporter green fluorescent protein (GFP) gene in the mouse embryo stem cell line R1. Methods: Three different kinds of GFP euko-expression vectors were constructed, and the expression efficiency was contrasted both at mRNA and protein levels after they were integrated into the chromosomes of host cells. Results: At protein level, the GFP expression level of the colonies transfected by the expression vector-pEF-GFP with the promoter of the peptide elongation factor (EF) were significantly higher than that of the colonies transfected by pCMV-GFP with CMV promoter and by pdCMV-GFP with double copies of CMV-GFP expression unit. There was no significant difference between the colonies transfected by pCMV-GFP and pdCMV-GFP. The detection results on mRNA level of GFP had the same tendency as that at protein level. Conclusion: (1) GFP gene expression efficiency controlled by EF promoter is distinctly higher than that by CMV promoter in NIH3T3 and R1 ES cell line.(2) A slight increase of the copy number of the foreign gene expression units in the host chromosome can not make obvious increase of its expression efficiency. (3) The vector express GFP in R1 ES cell line efficiently and stablely is obtained.