Carbon Monoxide ; analysis ; Spectroscopy, Fourier Transform Infrared ; methods

Air Pollutants, Occupational ; analysis ; Chromatography, Gas ; methods ; Indenes ; analysis ; Workplace

OBJECTIVETo evaluate the clinical results of the medial patellofemoral ligament (MPFL) reconstruction combined with the lateral retinacular release for the treatment of recurrent patellar dislocation.

METHODSFrom March 2011 to June 2013, 15 patients with recurrent patellar dislocation underwent arthroscopic MPFL reconstruction combined with the lateral retinacular release. The graft was autogenous semitendinosus and semimembranosus tendon. There were 5 males and 10 females with an average age of 19.4 years old (ranged,14 to 32 years old). The patients suffered recurrent patellar dislocation at least twice preoperatively. Preoperative conventional X-ray, CT, and MR examination were used to analyze the causes of the patellofemoral joint and MPFL injury. Preoperative Lysholm score was 69.85 ± 11.52. During operation, the arthroscopic examination was performed to evaluate the patellofemoral alignment and patellar tracking.

RESULTSAll the patients were followed up for an average of 27.6 months (ranged,12 to 36 months) with no recurrent dislocation and sub-dislocation. All the patients showed negative apprehension test at straight and 30 ° flexions of knee. The range of motion of knee returned to normal level at 12 months after operation. There were no patients with subjective discomfort of knee. Postoperative Lysholm score was improved to 92.60 ± 5.75.

CONCLUSIONThe technique of arthroscopic MPFL reconstruction combined with the lateral retinacular release is an effective surgical procedure for the treatment of recurrent patellar dislocation, which can relieve the symptom of knee and improve the patella stability and knee function.

Adolescent ; Adult ; Arthroscopy ; Female ; Humans ; Knee Joint ; surgery ; Male ; Patellar Dislocation ; physiopathology ; surgery ; Patellar Ligament ; physiopathology ; surgery ; Patellofemoral Joint ; physiopathology ; surgery ; Range of Motion, Articular ; Treatment Outcome ; Young Adult

This article aimed to study the antigenicity of nucleocapsid proteins (NPs) in six pathogenic phleboviruses and to provide theoretical evidence for the development of serological diagnostic reagents. NPs of six pathogenic phleboviruses were expressed and purified using a prokaryotic expression system and rabbits were immunized with individual recombinant NPs. Cross-reactions among NPs and rabbit sera were determined by both indirect ELISA and Western blotting analyses, and the sera titer was determined by indirect ELISA. Furthermore, sera from SFTS patients were also detected by each recombinant NP as a coating antigen using indirect ELISA. The cross-reactions and the sera titer were subsequently determined. Both the concentration and purity of recombinant NPs of six pathogenic phleboviruses met the standards for immunization and detection. The results of indirect ELISA and Western blotting showed that each anti-phlebovirus NP rabbit immune serum had potential serological cross-reactivity with the other five virus NP antigens. Furthermore, the sera from SFTS patients also had cross-reactivity with the other five NP antigens to a certain extent. Our preliminary study evaluated the antigenicity and immune reactivity of six pathogenic phleboviruses NPs and laid the foundation for the development of diagnostic reagents.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Cross Reactions ; Humans ; Nucleocapsid Proteins ; genetics ; immunology ; Phlebotomus Fever ; diagnosis ; immunology ; virology ; Phlebovirus ; classification ; genetics ; immunology ; isolation & purification ; Rabbits

BACKGROUND:Adiponectin possess functions of lowering blood glucose and blood lipids, and improve insulin sensitivity. But, controversy results about the effect of adiponectin on skeletal muscle have been reported.OBJECTIVE:To study the effects of eukaryon expressed adiponectin on the glycogen synthesis and glucose oxidation in skeletal muscle cell strain C2C12 myotubes by transfecting plasmids carrying mouse adiponectin.DESIGN: A controlled experiment.SETTING: The Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: PcDNA3.0 plasmid with mouse adiponectin cDNA, pcDNA3.0-mad (generously presented from Dr. Gong,University of Maryland), C2C12 cell strain (purchased from ATCC, GRL-1722), DMEM high glucose (Gibco), MEM (Hyclone), fetal bovine serum (Hanagzhou Sijiqing), equine serum (Hyclone), lipofectamine 2000 (Invitrogene), G418 (Gibco), rabbit anti-mouse adiponectin IgG (ACRP303-A, Alpha Diagnostic International), chemiluminescence kit (ECL+PLUS,Amersham), SABC instant immunohistochemistry kit (Boster), D-[U-14C] glucose (specific activity 9.25-13.32 GBq/mmol,NEC), scintillation fluid POP, POPOP (SIGMA), liquid scintillation counter (LS3801, Beckman, USA).METHODS:This study was carried out in the Central Laboratory of the Second Affiliated Hospital of Sun Yat-sen University from March to August, 2003. ① After extraction of plasmid, double digest with Xho Ⅰ and Xba Ⅰ and identification with HindⅢ digest were carried out. ② Plasmid pcDNA3.0-mad and pcDNA3.0 blank vector were transfected using liposome to C2C12 cells, and the stably transfected cells were screened by 500 mg/L G418 for 3 weeks, G418 resistant C2C12 cells were thereafter harvested, therefore stable transfected pcDNA3.0-mad and pcDNA 3.0 C2C12 cell strains were established.③ Adiponectin protein expression was determined by Western blot analysis and immunohistochemistry. ④ Glucose oxidation and glycogen synthesis detections were divided into control, vector and pcDNA3.0-mad (mad)group. Each group was further divided into 4 subgroups with 0, 0.5, 5 and 100 nmol/L insulin (n =6), respectively. Detection of glucose oxidation and glycogen synthesis was carried out with 14C-labeled glucose by counting radioactivity of 14CO2 or 14C labeled glycogen with scintillation, respectively.MAIN OUTCOME MEASURES:Changes of glycogen synthesis and glucose oxidation in skeletal muscle cell strain C2C12myotubes.RESULTS: ① Results of plasmid transfection and restrict digest: After plasmid extraction, double digest with Xba Ⅰ and Xho Ⅰ was carried out along with HindⅢ digest identification.Digest fragments were in accordance with expectation.Length of adiponectin cDNA fragment was 781 bp, plasmid fragment was 5 446 bp, adiponectin cDNA was inserted between digest sites (Xho Ⅰ and Xba Ⅰ ) of eukaryotic expression vector pcDNA3.0. ② Plasmid transfection of C2C12 cell and positive clone screening: On the 10th day of G418 media culture screening after transfection, most C2C12 cells died.Positive clone appeared at the 2nd week. G418 resistant C2C12 colonies were harvested at the 3rd week. ③ Western blot and immunohistochemical identifications: Both confirmed that adipoenctin gene was stably transfected into cells in the Mad group, with successful adipoenctin expression. ④ Effect of stably transfected adiponectin gene to myocyte glucose metabolism:The myocyte glycogen synthesis and glucose oxidation increased along with the increasing of insulin concentration. The linear regression analyses of measured myocyte glucose oxidation amount showed that the regression coefficients of the control group, blank vector group and mad group were 23.34, 2;3.23 and 26.06 respectively. This result indicated that in C2C12 cell stably transfected with adiponectin gene, when insulin concentration increased, the acceleration rate of glucose oxidation increasing was higher than other 2 groups. However, no significant difference could be observed in glycogen synthesis and glucose oxidation of C2C12 cells under basic status without insulin stimulus and treatment status with different insulin concentrations between control group, blank vector group and mad group (P＞ 0.05).CONCLUSION: ① We have successfully established stably adiponectin gene transfected C2C12 cell strain with adiponectin protein expression ability. ② Transfection with adiponectin cDNA had no significant effect on the glucose oxidation and glycogen synthesis of C2C12 myotubes.③ The glucose oxidation and glycogen synthesis of C2C12 myotubes increased with the increasing of insulin concentration. ④ Adipoenctin may coordinate with insulin in improving myocyte glucose oxidation and increasing myocyte glucose uptake.

BACKGROUND: Recent researches have indicated that the generation and development of retinoblastoma(RB) might also be related with other anti-oncogenes except the known Rb1 gene.OBJECTIVE: To explore the loci of other genes which possibly participated in RB generation and development and try to find and confirm the indicators for the loss of heterozygosity(LOH) with merits in surveillance and prognosis.DESIGN: A case analysis by employing RB patients as subjects SETTING: A center of gene diagnosis and therapy of a military medical university-affiliated hospitalPARTICIPANTS: The study was conducted in the center of gene diagnosis and therapy of Xinan Hospital affiliated to Third Military Medical University of Chinese PLA. Sixteen RB cases including 10 males and 6 females were the patients of the outpatient department of three-affiliated hospitals of the Third Military Medical University of Chinese PLA from May 1998 to October 2001.Inclusion criteria: in accordance with RB diagnostic criteria and younger than 3 years old; Exclusion criteria: family heredity history. Two eyes were involved in 12 cases and one eye was involved in 4 cases.METHODS: Fourteen micro-satellite DNA labels on the 13th chromosome in tumor or peripheral blood samples were separately amplified by polymerase chain reaction(PCR) to analyze the incidence of LOH of each locus. Simultaneously, the genetic source of loci loss was confirmed by genealogical analysis.MAIN OUTCOME MEASURES: Frequency of LOH incidence on the 13th chromosome.RESULTS: In 16 RB patients,LOH occurred in one or more than one locus on the 13th chromosome of 12 cases. Thereinto, the probability of LOH occurrence on three loci including D13S265,D13S263 and D13S153(in Rb1gene) was the highest. Ten loci of LOH in 12 LOH positive samples were confirmed from agnate chromosomes. The RB confirmation of LOH-positive group or LOH-negative group needed 504 days or 1086 days,which was significantly different(t=2. 357,P<0.05).CONCLUSION: RB confirmation was earlier in LOH-positive patients than LOH-negative patients. Except the confirmed Rb1 gene, LOH on two loci including D13S263(13q14.1-14.2) and D13S265 (13q31-32) also might have certain suggestive effect on early intervention and functional surveillance of RB patients.

Objective:To investigate the therapeutic effect of carbon dioxide laser tonsillectomy.Method:In this prospective,randomized study, One hundred and two patients were divided into laser group or control group. Patients of laser group were cured with carbon dioxide laser tonsillectom,and the control group was cured with routine method. All operations are executed by one person. Observation index included operation time, hemorrhage in operation, ache after operation, inflammatory reaction of raw surface, repair time of raw surface, rehaemorrhagia and scar.Result:Laser group had advantages of less operation time, less hemorrhage, less ache and less inflammatory reaction of raw surface. Laser group have hemorrhage in operation (7.2±2.1)ml, while control group have hemorrhage in operation (92.0±35.0)ml. Laser group have pseudomembrane early but desquamate late.Conclusion:Carbon dioxide laser tonsillectomy is effective to relieve pain, inflammatory reaction and with less time ,it's an safe , efficient and mini-trauma operation.

This study aims to investigate whether the nucleoprotein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV) can impact the cellular immunity of host cells. Gene segments that encode the NP and non-structural protein (NSs) of SFTSV were inserted into eukaryotic expression vector VR1012. Host proteins that interact with NP and affect immunity were identified with co-immunoprecipitation (IP), SDS-PAGE, mass spectrometry (MS), and Western blot. Co-localization of NP and the identified host proteins was confirmed by confocal microscopy. A 60kD SSA/Ro, a protein related to immunity, interacted with NP, as found by IP and MS. Confocal microscopy showed that NP and SSA/Ro were co-localized in cytoplasm. These results indicated that SFTSV NP may specifically bind to 60kD SSA/Ro and cause a series of immune responses and clinical symptoms.
Bunyaviridae Infections ; genetics ; metabolism ; virology ; HEK293 Cells ; Humans ; Nucleoproteins ; genetics ; metabolism ; Phlebovirus ; genetics ; metabolism ; Protein Binding ; Ribonucleoproteins ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

To establish a MacELISA method for the detection of IgM antibodies against Chikungunya virus (CHIKV), we prepared virus like particle (VLP) antigens of CHIKV using the whole structural protein C-E3-E2-6K-E1 encoding gene with a baculovirus expression system in Sf9 insect cells. The VLPs were purified and used to immunize Kunming mice. Then, polyclonal antibodies were purified from the samples of ascites with a protein G HiTrap SP column and labeled with horseradish peroxidase. A MacELISA method for the detection of IgM antibodies against CHIKV was assembled with goat anti-human IgM antibody, VLP antigens and an enzyme-labeled polyclonal antibody. The results were evaluated with a serum panel containing serum samples from laboratory-confirmed CHIK, HFRS patients, healthy donors, and commercially available CHIKV IgM as a quality control. It was shown that the MacELISA had a specificity of 99% (99/100), the coefficients of variation (CoV) within a plate were <10%, and the CoV of different ELISA plates in terms of the plate variation coefficient was <15%. A comparative analysis was performed to compare the current method against a commercial CHIKV IgM antibody detection kit for IIFA-IgM. The detection limit of MacELISA was significantly lower than that of the IIFA-IgM commercial kit (P< 0.0001). Here, we demonstrate that the VLP-based MacELISA is a promising tool for the early diagnosis and epidemiological investigation of CHIKV infection, with a high level of sensitivity and specificity for the detection of IgM antibodies against CHIKV.
Animals ; Antibodies, Viral ; blood ; Chikungunya Fever ; blood ; diagnosis ; virology ; Chikungunya virus ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin M ; blood ; Mice

To evaluate the adjuvant effect of recombinant enterovirus 71 (EV71) subunit vaccine formulated with chitosan, rabbits were orally immunized with recombinant VP1 (rVP1) or rVP1 mixed with chitosan adjuvant. Levels of virus-specific IgG and IgA antibodies in sera, mucosal wash buffer (intestine, nasal cavity, and lung), and feces were determined by indirect enzyme-linked immunosorbent assay (ELISA). The titers of neutralizing antibodies against EV71 were determined using cytopathic effect-based neutralizing assay, and levels of cytokines (IFN-gamma and IL-4) secreted from in vitro-cultured rabbit splenic lymphocytes under antigen stimulation were also determined by ELISA. Results showed that immunization with rVP1 alone could only induce low levels of serum IgG and mucosal IgA, while rVP1 combined with chitosan adjuvant were able to induce significantly higher levels of antibodies, rVP1 can only induce neutralizing antibodies when used in combination with chitosan. Levels of IFN-gamma and IL-4 in the group immunized with rVP1 plus chitosan were significantly higher than those in the group immunized with rVP1 only or those in the control groups. Our study lays the foundation for development of oral VP1 vaccine against EV71 infection.
Adjuvants, Immunologic ; administration & dosage ; Animals ; Antibodies, Viral ; immunology ; Chitosan ; administration & dosage ; immunology ; Enterovirus A, Human ; genetics ; immunology ; Enterovirus Infections ; immunology ; prevention & control ; virology ; Female ; Humans ; Rabbits ; Vaccination ; Vaccines, Subunit ; administration & dosage ; genetics ; immunology ; Viral Proteins ; administration & dosage ; genetics ; immunology ; Viral Vaccines ; administration & dosage ; genetics ; immunology