Objective To establish RT-PCR-RFLP method for studying the genotype of wild mea-sles virus strains isolated from Tianjin area from 2002 to 2008. Methods Isolations of measles virus were carried out by tissue culture method from urine and throat swab specimens collected from suspected cases. RNA were extracted from the virus specimens. The 594 bp fragment of C terminal of the N (nucleoprotein) gene was amplified by one-step RT-PCR, then the PCR products were digested with Bcn I , separated on agarose gel electrophoresis and then analyzed by the method of RFLP (restriction fragment length polymor-phism). In addition, above results were compared with DNA sequencing. Phylogenetic tree was plotted based on the results for the genetic relationship and distance analysis. Results Sixty-nine measles virus strains were isolated from 189 specimens from 2002 to 2008, of which the C terminals of N gene were all de-tected positive. Among the 69 strains of measles virus isolates, 98.55% (68/69) belonged to Hla sub-geno-type which was the predominant sub-genotype, and only one strain (1.45%) belonged to H1b sub-genotype by RFLP analysis which was in accordance with the results by DNA sequencing method. Phylogenetic tree analysis indicated the H1a sub-genotype measles virus strains should be further divided into 2 clades, and the variation fluctuated between 0.2% and 3.8%. There were transmission chains caused by different virus strains co-cireulation. Conclusion A genotype, H1a and H1b sub-genotype can be identified by RT-PCR-RFLP assay specically based on the restriction enzyme Bcn I .The RT-PCR-RFLP assay can be a rapid, simple, accurate and efficient method for large-scale surveillance of measles virus strains in China.

This study examined the effect of GHRP-6, a known GHSs receptor agonist, on the phosphorylation of cAMP-responsive element-binding protein (CREB) and the underly mechanism. GH3 cells were cultured and subjected to different treatments as follows: GHRP-6, GHRP-6 plus GHRH, phorbol ester (PMA), an activator of PKC, alone or in combination with GHRP-6, Gö6983, a general inhibitor of PKCs, in the presence or absence of GHRP-6, rottlerin, an inhibitor of PKCs, alone or plus GHRP-6. The cells were transiently transfected with PKCsigma-specific siRNA and then treated with GHRP-6. GH level was measured by enzyme-linked immunosorbent assay (ELISA). The expression of phosphor-CREB, PKCsigma, PKCtheta and phosphor-PKCsigma was determined by Western blotting. The results showed that GHRP-6 stimulated GH secretion in both time- and dose-dependent manners and enhanced the effect of GHRH on GH secretion. GHRP-6 was also found to induce CREB phosphorylation. Moreover, GH secretion was enhanced by the PKC activator PMA and reduced by the PKC inhibitors (Gö6983, rottlerin) and knockdown of PKCsigma. PKCsigma could be activated by GHRP-6. It is concluded that PKC, especially PKCsigma, mediates CREB phosphorylation and GHRP-6-induced GH secretion.

Objective To study the value of the diameters of fetal umbilical artery and umbilical vein in early prenatal screening program for thalassemia by ultrasound.Methods A total of 120 cases of singleton pregnant with thalassemia at 12- 28 gestational weeks were detected by ultrasound,328 cases of normal singleton pregnant at 12 - 28 gestational weeks were selected as controls.The diameters of fetal umbilical artery and umbilical vein were tested and compared between the two groups.The diagnostic values were evaluated by ROC curve.Results At 12 - 15+6 gestational weeks,the diameters of umbilical artery and umbilical vein of thalassemia group were larger than those of control group(P ＜0.05).At 16 - 19+6 gestational weeks, the diameter of umbilical vein of thalassemia group was larger than that of control group,the differences were significant (P ＜0.05).The cutoff threshold of the diameter of fetal umbilical artery was 1.80 mm,the diameter of fetal umbilical vein was 2.95 mm at 12 - 15+6 gestational weeks,the diameter of fetal umbilical vein was 3.80 mm at 16- 19+6 gestational weeks,which were obtained by ROC analysis.The sensitivity of this cutoff threshold to identify thalassemia were 83.3％, 83.3％, 47.8％,respectively, while the specificity were 90.5 ％, 95.2 ％, 85.4％, respectively.Conclusions The diameters of fetal umbilical artery and umbilical vein detected by ultrasound have great value on early prenatal screening of thalassemia, these indexes can be used as an important factors in early prenatal screening of thalassemia.

OBJECTIVE To discover diversity of X gene sequences of hepatitis B virus isolates in hepatitis B induced liver cirrhosis patients and HBV carriers.METHODS DNA fragment including X gene sequences of hepatitis B virus was amplified,sequencing PCR products was preformed.The PCR products of three liver cirrhosis patients and three HBV symptomless carriers were cloned into pGEMT Easy vectors.Positive clones with target sequences were selected out for sequencing.Sequence comparison was made to find the identity.RESULTS A comparison of T1762/A1764,G1719T,T1727G/A,G1730C and T1753C mutations in a core promoter between the liver cirrhosis patients and the HBV carriers showed that the HBV isolates from the former had higher frequencies of mutation than the latter.The X promoter region of the HBV isolates from the liver cirrhosis patients showed higher frequencies of mutation than the isolates from the HBV carriers.Additionally,the homology between clones of X gene from one individual with liver cirrhosis averages 91.3-99.7%,that of HBV carriers averages 96-100%.CONCLUSIONS The core and X promoter region of the HBV isolates from the liver cirrhosis patients show the higher frequencies of mutation than the isolates from the HBV carriers.There are HBV quasispecies which possess great variation in the liver cirrhosis patients.

OBJECTIVESTo evaluate the specificity of three-color flow cytometry in childhood acute lymphoblastic leukemia (ALL) immunophenotyping.

METHODSImmunophenotyping was performed by three-color flow cytometry analysis using CD(45)/SSC gating.

RESULTSThe percentage of blasts was correlated better with leukemic cell count compared with that of FSC/SSC, and the false positive results were low. Among eighty six cases of ALL, 95.3% was B-ALL, in which common-ALL and Pro-B-ALL were 76.8% and 6.1%, respectively, and 2.3% was T-ALL. CD(34)(+) and myeloid-associated antigen expression were observed in 57.0% and 34.9% of the cases, respectively, among which Pro-B-ALL was the commonest. CD(33) was more commonly expressed than CD(13) in Pro-B-ALL cases, but no difference in the expression between these two antigens in other subtypes.

CONCLUSIONGating of CD(45)/SSC eliminated effection of normal cells to blasts in bone marrow, with which the immunophenotyping results were more reliable.

Antigens, CD ; analysis ; Child ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology

Objective To determine if there is any difference in neuronal and glial(astrocytic and microglial)activation in the spinal cord in three rat models of neuropathic pain.Methods Twenty-four SD rats weighing 150-200 g were randomly divided into 4 groups(n=6 each):Ⅰ control group;Ⅱ chronic constrictive injury group(CCI);Ⅲ spinal nerve ligation group(SNL)and Ⅳ spared nerve injury(SNI).No operation was performed in control group.In CCI group left sciatic nerve was exposed and loosely ligated with catgut.In SNL group the L_5 spinal nerve was exposed and ligated with silk suture and cut.In SNI group tibial nerve and common fibular nerve were ligated and cut.Pain threshold was measured using plantar tactile stimulator(Ugo,Basile Co. Italy)every other day from 3 days before until 15 days after operation.50% paw withdrawal threshold was measured using up-and-down sequential mechanical stimulation of different intensity(0.45,0.70,1.20,2.00, 3.63,5.50,8.50,15.10 g)applied to the plantar surface of the injured paw.On the 15~(th) day after operation after pain threshold was measured the animals were anesthetized with intraperitoneal 10% chloral hydrate 400 mg? kg~(-1).The L_(5,6) segment of the spinal cord was isolated.Neuronal,astrocytic and microglial activation was determined by immuno-histochemistry with antibodies of c-Fos(a proto-oncogene protein),GFAP(an astrocyte marker)and OX-42(a microglial marker).Results The 50% paw withdrawal threshold reached the lowest level on the 7~(th) day after operation.The lowest level was maintained until the 15~(th) day after operation in group CCI,SNL and SNI.The 50 % paw withdrawal threshold was(14.1+1.5)g in control group,(2.5+0.5)g in CCI group, (1.5?0.6)g in group SNL and(0.8?0.4)g in group SNI.The number of c-Fos positive neurons in laminae Ⅳ-Ⅵ of dorsal horn was significantly greater in group CCI,SNL and SNI than in control group,but there was no significant difference among the 3 peripheral nerve injury groups.The activation of astrocytes and microglias in laminae Ⅰ-Ⅳ of dorsal horn was significantly increased in group CCI,SNL and SNI than in control group but there was no significant difference among the 3 peripheral nerve injury groups.Condusion There is no significant difference in activation of neurons and astrocytes and microglias in the ipsilateral dorsal horn among the 3 pain models.

Animals ; Dynorphins ; metabolism ; Parkinson Disease ; drug therapy ; metabolism ; Peptides ; pharmacology ; Rats ; Scorpion Venoms ; pharmacology

OBJECTIVETo assess the influence of different hinge positions on clinical results of expansive open-door laminoplasty (EOLP) for cervical spondylotic myelopathy (CSM).

METHODSA total of 102 CSM patients who underwent EOLP from February 2006 to February 2007 were enrolled in this randomized controlled trial. Using a random digits table, 57 patients with the hinge located at the inner margin of the lateral mass were classified as wide-open group, while 45 patients with the hinge positioned at the lamina margin served as narrow-open group. All patients were observed over 24 months, and the clinical and radiological results were analyzed statistically.

RESULTSThere were no significant differences in operation duration, intraoperative bleeding volume, Japanese Orthopaedic Association (JOA) scores, cervical curvature index, range of motion and neural function recovery rate. The neural functions were satisfactorily improved after surgery in both groups, while the severity of axial symptoms was significantly lower in the narrow-open group than in the wide-open group (P equal to 0.003). The incidence of C(5) palsy in the wide-open group was higher than that in the narrow-open group (5.3% vs 0), even though the difference did not reach statistical significance (one tailed Fisher's exact test, P equal to 0.17).

CONCLUSIONSProper inward shift of the hinge can ensure effectiveness of surgical decompression, avoid an excessive backward shift of the spinal cord, decrease the incidence of C(5) palsy and alleviate the severity of axial symptoms.

Aged ; Female ; Humans ; Laminectomy ; methods ; Male ; Middle Aged ; Spondylosis ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed

OBJECTIVETo evaluate the efficacy of Chinese medicine (CM) combined adjuvant chemotherapy in postponing relapse and metastasis of radical resected Ib-IIIa stage non-small cell lung cancer (NSCLC) patients, and to explore its effect in improving their quality of life (QOL) and clinical symptoms.

METHODSWe designed a cohort study of 336 radical resected Ib-IIIa NSCLC patients by analyzing disease free survival (DFS) using Log-rank test. They were randomly assigned to the control group (155 cases, treated by adjuvant chemotherapy group) and the test group (181 cases, treated by adjuvant chemotherapy combined CM). By using controlled method, 60 radical resected NSCLC patients undergoing NP/NC program in 2012 (vinorelbine 25 mg/m2, combined with cisplatin 75 mg/m2 on day 1 and day 8/on day 1 or on day 1, 2, and 3; or carboplatin AUC = 5 on day 1) were assigned to the control group (29 cases) and the test group (31 cases). QOL scores (using EORTC QLQ-LC43 questionnaire) and TCM symptoms scores were compared between the two groups before chemotherapy, peri-chemotherapy (one day before the 2nd course of chemotherapy) , and after chemotherapy (20 days after ending the 4th course of chemotherapy).

RESULTS(1) The median DFS was longer in the test group than in the control group, but with no statistical difference between the two groups (42.73 months vs 35.57 months , P = 0.179). In the subgroup analysis, there was statistical difference in IIIa stage DFS. The median IIIa stage DFS of was longer in the test group than in the control group with statistical difference (27.87 months vs 19. 93 months, P = 0.047). (2) In the control study, repeated measured data indicated there was significant difference in physical functions between the two groups (P < 0.05). Total scores for health states decreased more in the test group than in the control group, but with no statistical difference (P > 0.05). Scores for constipation and CM syndrome scores were higher in the test group than in the control group (P < 0.05).

CONCLUSIONSCM had advantages in postponing DFS of radical resected NSCLC patients, especially in IIIa stage. CM could improve their QOL and clinical symptoms during adjuvant chemotherapy.

Adjuvants, Immunologic ; Adjuvants, Pharmaceutic ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carboplatin ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Chemotherapy, Adjuvant ; Cisplatin ; therapeutic use ; Cohort Studies ; Disease-Free Survival ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lung Neoplasms ; Quality of Life ; Vinblastine ; analogs & derivatives ; therapeutic use

Objective To explore the correlated factors and clinical features of cognitive impariment in parkinson's disease (PD).Methods A total of 419 patients with PD were collected from Xiangya Hospital of Centre-South University during Mar 1st,2017 to Nov 30th,2017.The cognitive functions of patients were assessed with the Mini-Mental State Examination (MMSE),and the basic information and the motor symptoms of 419 PD patients were selected at the same time.The PD patients were classified into three groups according to the MMSE score:PD with no cognitive impairment (PD-NC),mild cognitive impairment in PD (PD-MCI),and Dementia in PD (PD-D).The data were analyzed by SPSS 20.0.Results There were 156 patients with PD-MCI (37.2％) and 64 patients with PD-D (15.3％).The difference of sex and disease duration among three groups were not statistically significant (P ＞ 0.05).The significant difference was found among PD-D,PD-MCI,and PD-NC groups in age of onset,age,educational attainment,Unified Parkinson's disease Rating Scale (UPDRS)-Ⅱ score,UPDRS-Ⅲ score and Hoehn-Yahr stage (P ＜ 0.05).There were significant differences among three groups in MMSE score and its items (P ＜ 0.01).Logistics regression analysis found that the age of onset,educational attainment,and Hoehn-Yahr stage were the risk factors of cognitive impairment in PD patients (P ＜ 0.05).Conclusions Cognitive impairment is common in PD patients,and it is relevant to the age of onset,educational attainment and the severity of illness of PD patients.