Objective To assess the new edition of WHO Japanese Encephalitis (JE) Surveillance Standards (WHO Standards) based on syndrome surveillance data and to provide field evidence regarding the standards. Methods Based on syndrome surveillance data, acute encephalitis syndrome (AES) case was categorized, according to the WHO Standards. A cohort study was applied to estimate the AES definition set in the Standard and relative risk was computed to estimate the existence and intensity of statistical correlation between AES and JE cases. Percentage of attributable risk was counted to describe the coverage of AES for JE cases in the studied population. Sensitivity,specificity, Youden index and positive predictive value of AES components were calculated for the purpose of identifying the clinical values under the screening program. Results 1424 suspected cases were evaluated in the surveillance program and 1396 cases with ELISA result, of which 109 positive cases were detected. According to the "standardized" classification, a total of 706 cases in line with AES case deftuition, were categorized into 83 cases of JE, 425 cases of AES unknown and 198 cases of AES other agent. In the cohort study,a relative risk of 4.62 (95% CI:2.80-7.63 ) and the percentage of attributable risk as 78.35% (95% CI: 64.25% -86.89% ) were observed. Conclusion The AES definition for JE was significantly effecting on the screening programs and a strong correlation strength was observed in the study. AES syndrome could cover most of the JE cases. "Convulsions",with appreciative screening value, was recommended to be involved into the new version of the WHO Standards.

OBJECTIVETo investigate therapeutic effects of stiletto needle and Dichofenac Diethylammon in treating knee osteoarthritis pain at early and medium-term.

METHODSSixty-seven patients with early and medium-term knee osteoarthritis were randomly divided into two groups, 34 cases in stiletto needle group (treatment group) and 33 cases in Dichofenac Diethylammon (control group). Among them, 60 patients finished treatment and followed-up for 1 months (7 cases fall off totally, 4 cases in treatment group and 3 cases in control group). In treatment group, there were 5 males and 25 females with a mean age of (55.90 +/- 9.34) years (ranged, 40 to 68 years), the course of disease for 0.25 to 1 year, mean (0.87 +/- 0.34) years, treated by stiletto needle once a week, and 3 weeks were a course. In control group, there were 6 males and 24 females with a mean age of (58.67 +/- 7.39) years (ranged, 40 to 70 years), the course of disease for 0.25 to 2 years, mean (0.93 +/- 0.60) years, treated by Dichofenac Diethylammon, three times a day for 3 weeks. Soft tissue displacement and banana area were tested by soft tissue tension tester, pressure value were measured by pressure measuring instrument, VAS score were recorded by pain visual analog scale record, HSS scoring and effects were used to evaluated and statistical analyzed.

RESULTSThere were significant differences in tissue displacement, banana area, pressure value, VAS scoring and HSS scoring between two groups before and after treatment, and tissue displacement, pressure value, HSS scoring in treatment group were higher than control group, while banana area and VAS scoring were lower than control group. According to HSS scoring and VAS scoring, 3 cases were healed, 8 cases were markedly improved, 18 cases were effective and 1 case was effectiveless in treatment group;while 1 case was healed, 4 cases were markedly improved, 20 cases were effective and 5 cases were effectiveless in control group. There was no significant meaning in therapeutic effects.

CONCLUSIONStiletto needle for knee osteoarthritis pain can effectively reduce high tension of local soft tissue ,alleviate symptom of pain, and improve knee function.

Adult ; Aged ; Biomechanical Phenomena ; Case-Control Studies ; Diclofenac ; analogs & derivatives ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Needles ; Osteoarthritis, Knee ; complications ; Pain ; complications ; drug therapy ; pathology ; physiopathology ; Pain Management ; methods ; Treatment Outcome

OBJECTIVETo investigate the prosthesis selection, precaution and curative effect of total knee replacement in severe gonarthrosis.

METHODSFrom January 1996 to July 2009, 50 patients (58 knees) with severe gonarthrosis underwent total knee replacement, included 12 males and 38 females, aged from 46 to 80 years with an average age of 66.5 years old. Six artificial hinged knee joints, 37 general knee joints and 15 ScorpioNRG knee joints were used according to the deformity of the knees. Fifty patients (58 knees) were followed up and evaluated with the HSS (the Hospital for Special Surgery Knee Score) scoring system.

RESULTSThe duration of follow-up ranged from 6 months to 8 years (means 62 months). According to the HSS scoring system, the mean preoperative score was (38.6 +/- 8.76) points, and the postoperative score was (86.50 +/- 9.45) points. The clinical effect was excellent in 20 cases (23 knees), good in 28 cases (32 knees),and acceptable in 2 cases (3 knees).

CONCLUSIONStable and functional recovery knee joint without deformities and pains could be obtained after total knee replacement by carefully chosing of prosthetic replacements according to the different degrees of joint deformity of severe gonarthrosis.

Aged ; Arthroplasty, Replacement, Knee ; Female ; Follow-Up Studies ; Humans ; Joint Deformities, Acquired ; surgery ; Knee Joint ; pathology ; Male ; Middle Aged ; Osteoarthritis, Knee ; pathology ; surgery

ObjectiveTo establish a novel assay for HIV-1 p24 ultrasensitive detection based on Gold Nanoparticle Probe (GNP) and PCR.MethodsSandwich ELISA method was established by a pair of anti-p24 monoclonal antibodies (mAbs),1G12 and 1D4,and was used to detect recombinant HIV-1 p24 antigen.The bio-barcode DNA was 47 bp,selected from genome of Arabidopsis,and formed double-stranded DNA by hybridization with the capture DNA (complementary with bio-barcode DNA) modified with sulfhydryl.Then double-stranded DNA were conjugated on the surface of 1D4-modified gold nanoparticles by sulfhydryl,and the Gold Nanoparticle Probe was produced.1G12 was precoated in the micropaltes,and in the presence of target recombinant HIV-1 p24 protein,a sandwich immuno-complex would form by adding GNP.Then the bio-barcode DNA in the immuno-complex were released by heating as detection signal,and consequently characterized by the polymerase chain reaction (PCR) with synthesized special primers and analyzed by 4％ agar gel electrophoresis,so HIV-1 p24 antigen could be evaluated.The sensitivity comparison between the new assay and ELISA can be done.ResultsSandwich ELISA was used to quantify HIV-1 p24 antigen by monoclonal antibodies 1G12 and 1D4,and the limit of detection (LOD) was 1000 pg/ml.The new GNP assay was established by the same pair of antibodies,combined with PCR and agar gel electrophoresis,and was used to indirectly detect HIV-1 p24 antigen.The band intensity of PCR products paralleled with the quantity of HIV-1 p24 antigen,and the limit of detection (LOD) could reach down to 1 pg/ml.ConclusionThe new assay based on GNP and PCR was efficient in the detection of HIV-1 p24,which is at least 3 orders of magnitude more sensitive than traditional ELISA.

This study was aimed to investigate the expression of c-MPL in acute myeloid leukemia (AML) and the correlation of the c-MPL expression with CD34 and CD38, so as to define the expression of c-MPL in leukemic stem cells. The expression levels of CD34, CD38 and c-MPL were detected by flow cytometry in bone marrow cells from 29 newly diagnosed AML patients. The relationship of c-MPL positive cell ratio with clinical parameters and correlation of c-MPL with CD34 and CD38 expression in AML patients were analyzed. The results showed that expression level of c-MPL in AML patients was significantly higher than that of normal controls (P < 0.05), and the expression level of c-MPL did not correlate with age, sex, white blood cell count, AML1-ETO fusion gene and remission after chemotherapy, but the expression of c-MPL in M2 and M5 patients was higher than that of normal control (P < 0.05). Expression of c-MPL in CD34 positive AML patients was obviously higher than that in CD34 negative AML patients (P < 0.01). c-MPL was significantly higher expressed in CD34(+) cells than that in CD34(-) cells (P < 0.001), while c-MPL expression was not significantly different between CD34(+)CD38(-) and CD34(+)CD38(-) cell groups. Positive correlation between c-MPL and CD34 expression was observed (r = 0.380, P = 0.042). It is concluded that expression of c-MPL is higher in AML patients, and positively correlates with the expression level of CD34. The c-MPL expresses in leukemic stem cells.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Female ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Male ; Middle Aged ; Neoplastic Stem Cells ; metabolism ; Receptors, Thrombopoietin ; metabolism ; Young Adult

OBJECTIVETo investigate the inhibitory effects of intervention of the tumor necrosis factor-alpha (TNFa)/nuclear factor-kappa B (NF-kappaB) signaling pathway activation on hepatoma cell proliferation and to explore its mechanism.

METHODSA rodent hepatoma model was established by feeding N-2-fluorenylacetamide (2-N-FAA) to male Sprague-Dawley rats. Human subjects with various liver diseases were enrolled in the study, and serum and peripheral blood nuclear cells were collected for analysis. HepG2 cells were cultured in vitro and treated with anti-TNFa (monoclonal antibody, mAb) to down-regulate its expression or transfected with siRNA targeting the p65 subunit of NF-kappaB to inhibit its activation. The liver cell line L02 was used as a control. Changes in protein and gene expression levels of NF-kappaB and TNFa were analyzed by Western blotting or enzyme-linked immunosorbent assay and real-time PCR, respectively. Changes in the cell cycle or apoptosis were evaluated by flow cytometry or Annexin-V/PI double-labeling assay, respectively.

RESULTSTNFa and NF-kappaB expression showed increasing trends during the malignant transformation of rat hepatocytes, and the differential expression patterns showed association with histopathological alterations in the hepatocytes. Following treatment with the TNFa mAb, the HepG2 cells showed a higher percentage of apoptotic cells than the untreated control cells (21.45% +/- 4.07% vs. 5.63% +/- 0.93%, q =10.07, P less than 0.01).There was a significant difference in the rate of cells in the G0/G1 phase in the p65-siRNA transfected cells (66.23% +/- 1.29% vs. untreated control cells: 59.00% +/- 1.02%, q =10.98, P less than 0.01). The decreased expression of TNFa and NF-kappaB in cell culture supernatants was positively correlated with the dose of treatment (r =0.89, P less than 0.01), with the most robust decreases being achieved with the highest concentrations ( P less than 0.01). NF-kappaB expression was significantly higher in the HepG2 cells than in the L02 cells, and transfection of p65-siRNA reduced the mRNA (93%) and protein (62%) levels and increased the cell apoptosis index (to 85%).

CONCLUSIONProliferation of hepatoma cells may be significantly inhibited by intervening in the activation of the TNFa/NF-kappaB signaling pathway, which promotes cell apoptosis and blocks cell cycling.

Adult ; Aged ; Animals ; Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Cell Proliferation ; Female ; Hep G2 Cells ; Hepatocytes ; metabolism ; Humans ; Liver Neoplasms ; pathology ; Male ; Middle Aged ; NF-kappa B ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Transfection ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo measurement the vertebra morphology using multi-planar reformations method of multi-slice spiral CT (MSCT) in AIS girls, and To compare with age and gender-matched controls in order to affirm the morphology results of abnormal development of the anterior and posterior elements in AIS.

METHODSThoracic and lumbar spine multi-slice spiral CT was undertaken on 52 girls with AIS between the age of 10 and 18 years from June 2004 to May 2008 in Peking Union Medical College Hospital and Beijing Shijitan Hospital, and 54 age and gender-matched non-IS controls. Multiple measurements (including the anterior column and posterior column) of each thoracic and lumbar vertebra were obtained using the 3D-MPR technique. The patients and control were divided into 10-14 years old group and 15-18 years old group. The corresponding vertebral anterior height, vertebral posterior height, transverse distance, vertebra central width, vertebra anterior-posterior distance, area of pedicle, pedicle height, pedicle width, and upper-lower facet distance were compared.

RESULTSCompared with the controls, the 10-14 years old group girls' spine had longer anterior column height, relative shorter posterior column (there are difference from thoracic 2-11 and lumbar 1-3, P < 0.05), relative longer anterior-posterior vertebral distance, vertebral wedging changes in vertebral, distinct vertebral and pedicle asymmetry on the concave and convex side, and upper-lower facet distance asymmetry on the concave and convex side. The difference between the anterior and the posterior column ratio was significantly different from that in the controls (P < 0.01). But there hadn't the same difference in 15 - 18 years old group.

CONCLUSIONSThere are differences in coronal plane vertebra wedging changes in AIS patient. There are high vertebra height, relative shorter posterior column, relative longer anterior-posterior vertebral distance, and relative slender vertebra in 10 - 14 years old AIS patients. This may lead to the initial and progression of scoliosis.

Adolescent ; Case-Control Studies ; Child ; Female ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; Scoliosis ; diagnostic imaging ; surgery ; Thoracic Vertebrae ; diagnostic imaging ; Tomography, X-Ray Computed

OBJECTIVETo observe the expression and effect of human calcitonin gene-related peptide (hCGRP) gene mediated by recombinant adeno-associated virus (rAAV) in primary cultured corporal cavernosum smooth muscle cells of the rat and explore the possibility of using CGRP gene for gene therapy in erectile dysfunction.

METHODSThe primary cultured corporal cavernosum smooth muscle cells of the rat were randomly divided into 4 groups and infected with recombinant virus VssHGCMV-hCGRP, VssHGCMV, VssC-MV-GFP and the untreated, respectively. CGRP-like immunoreactivity was measured by protein dot blot assay in the 24 h-culture medium, and intracellular cAMP and cGMP levels in the cultured cells were also determined using radioimmunoassay to ascertain bioactivity of transduced CGRP.

RESULTSThe exogenous gene was transferred into primary corporal cavernosum smooth muscle cells by VssHGCMV-hCGRP infection and efficiently expressed. Compared with the control group, intracellular cAMP level in the cell infected by VssHGCMV-hCGRP was significantly increased (48.7 +/- 1.1 nmol/L vs 7.8 +/- 1.4 nmol/L, P < 0.01), whereas cGMP level remained unchanged in two groups, and CGRP-like immunoreactivity was also detected in the culture medium infected by VssHGCMV- hCGRP.

CONCLUSIONThe system of secretory expressing bioactive peptide rAAV mediated gene transfer may be used to express efficiently exogenous gene in corporal cavernosum smooth muscle cells and affect cAMP level in the corporal cavernosum smooth muscle cells of the rat.

Animals ; Calcitonin Gene-Related Peptide ; biosynthesis ; genetics ; Cells, Cultured ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Dependovirus ; genetics ; Male ; Muscle, Smooth ; cytology ; metabolism ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley ; Recombination, Genetic ; Transfection

OBJECTIVETo determine whether recombinant adeno-associated virus-mediated overexpression of hCGRP in the corpus cavernosum can affect the continuous production of hCGRP in the penile tissue and enhance erectile responses in STZ-induced diabetic rats.

METHODSDiabetes mellitus was induced by a single intraperitoneal injection of 60 mg/kg streptozotocin in male SD rats. VssHGCMV-hCGRP, VssCMV-GFP and rAAV solution were injected into the corporal cavernosum of STZ-induced diabetic rats, respectively. The corporal tissue was obtained from groups of 8 rats on day 5 post-injection, and the expressions of CGRP and GFP in cavernosal tissue were detected using immunohistochemistry and laser scanning confocal microscopy, respectively. Cavemosal tissue cAMP and cGMP levels were measured using radioimmunoassay. On day 5 post-injection, intracavernous pressure induced by electrostimulation of penile dorsal nerves was measured and recorded with a biological signal processing system in each group rat.

RESULTSrAAV transduction efficiency of GFP reporter gene was measured by laser scanning confocal microscopy and was observed in the penile tissue, especially in the corporal cavernosum and the vessel 5 days after transfection with VssCMV-GFP. Immunohistochemistry showed that the CGRP increased in the corporal cavernosum. In addition, both cAMP and cGMP levels in the corpora cavernosa transfected with VssHGCMV-hCGRP were significantly increased, compared with controls [(48.4 +/- 6.5) nmol/L and (21.2 +/- 13.6) nmol/L vs (16.7 +/- 2.5) nmol/L and (0.42 +/- 0.12) nmol/L, respectively]. More importantly, 5 days after administration of VssHGCMV-hCGRP,a significant increase was observed in the erectile response to penile dorsal nerve stimulation in the diabetic rat [(60.5 +/- 4.5) mm Hg vs (22. 3 +/- 1.3) mm Hg].

CONCLUSIONSThis results demonstrate that rAAV-mediated transfer of the CGRP gene can increase production of endogenous CGRP, cAMP and cGMP in corpora cavernosa of STZ-induced diabetic rats. Moreover, overexpression of CGRP enhances ICP and the erectile response to penile dorsal nerve stimulation in the diabetic rat.

Adenoviridae ; genetics ; Animals ; Calcitonin Gene-Related Peptide ; biosynthesis ; genetics ; Diabetes Mellitus, Experimental ; physiopathology ; Green Fluorescent Proteins ; biosynthesis ; Humans ; Male ; Penile Erection ; physiology ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transfection

Hepatitis B virus (HBV) infection is an important global public health concern and a major cause of chronic hepatitis, cirrhosis and liver cancer. Many studies have shown that different genotypes and subtypes have significant differences in pathogenicity, thus affecting the disease progression and prognosis of infected individuals. So far, a total of 10 HBV genotypes and more than 40 subtypes have been reported across the world, and these subtypes have shown distinct distribution characteristics. In the present review, we systematically summarized the current situation on the global distribution of HBV genotypes.