Objective:To obtain monoclonal antibodies(McAb) against Staphylococcal enterotoxin C2(SEC2) and establish method for detecting SEC2.Methods:The secreted SEC2 from staphylococcus aureus was used as antigen to immune BALB/c mice. Monoclonal antibodies against SEC2 were prepared by normal hybridoma technique. By identifying the characters of McAbs, the quantitative detection ELISA test method were established and were preliminarily applied.Results:Four hybridmas producing antibodies against SEC2 were obtained. IgG isotypes of four McAbs were IgG1. Their binding site was different except two McAbs that shared the same binding site.The McAbs were proved to be specific for SEC2.The sandwich ELISA method had good specificity, sensitivity and reproducibility, and it was founded to be able to detect SEC2 at concentration from 0.5 to 20 ng/ml. Its recovery ranged between 97.8% and 101%,and CV value ranged between 2% and 5%.Conclusion:The prepared McAb against SEC2 can be used for SEC2 immunoassay. This work provides a basis for controlling the quality of JINPUSU and researching staphylococcal enterotoxin.

OBJECTIVE: To investigate the psychological statuses of Chinese adults with moderate-to-severe persistent allergic rhinitis, and evaluate the effects of nasal symptoms on their psychological statuseses. METHOD: The Symptom Checklist-90 (SCL-90) or self-reporting Inventory was employed to analyze the psychological statuses of 539 adults with moderate-to-severe persistent allergic rhinitis. RESULT: The SCL-90 scores of the adults with moderate-to-severe persistent allergic rhinitis were statistically higher than those of non-allergic adults in terms of somatization, depression, anxiety and hostility. No statistical discrepancies existed in gender or age. The course of disease contributed to somatization and compulsion. The effects of nasal symptoms included two aspects: nasal obstruction had a conspicuous impact on somatization, compulsion, interpersonal sensitivity, depression, anxiety and psychosis, while nasal itching contributed to somatization, depression and anxiety. CONCLUSION The psychological statuses of adults with moderate-to-severe persistent allergic rhinitis is evidently worse than that of non-allergic adults. Symptoms such as nasal obstruction and itching had an obvious impact on outpatients' psychological statuses.
Adult ; Humans ; Nasal Obstruction ; Rhinitis, Allergic, Perennial ; diagnosis ; psychology

Objective: To investigate the prevalence and influencing factors of depressive symptoms among grassroots healthcare workers in Zhejiang Province, so as to provide insights into improving their mental health. Methods: Grassroots healthcare workers of community health service centers and township health centers were sampled from one county (city, district) in each of 11 cities in Zhejiang Province using a convenience sampling method from December 2022 to January 2023. Participants' gender, age, educational level and average daily sleep duration in the past week were collected through questionnaires, and depression symptoms was investigated according to Self-rating Depression Scale. Factors affecting the depressive symptoms were identified using a multivariable logistic regression model. Results: A total of 1 946 questionnaires were distributed, and 1 945 valid questionnaires were recovered, with an effective response rate of 99.95%. There were 444 boys, accounting for 22.83%, and 1 501 girls, accounting for 77.17%. The median age was was 36 (interquartile range, 44) years. There were 786 healthcare workers detected with depressive symptoms, with a prevalence rate of 40.41%, and the prevalence rates of mild, moderate and severe depressive symptoms were 26.94%, 10.49% and 2.98%, respectively. Multivariable logistic regression analysis identified age (50 years and older, OR=0.572, 95%CI: 0.386-0.846), annual income (100 000 to 149 999 Yuan, OR=0.780, 95%CI: 0.635-0.958; 150 000 to 199 999 Yuan, OR=0.463, 95%CI: 0.282-0.760; 200 000 Yuan and above, OR=0.303, 95%CI: 0.098-0.937), vocation (nurse, OR=1.593, 95%CI: 1.252-2.027) and sleep duration (less than 7 hours, OR=2.164, 95%CI: 1.768-2.648) as factors affecting depressive symptoms among grassroots healthcare workers in Zhejiang Province. Conclusions The prevalence of depressive symptoms among grassroots healthcare workers in Zhejiang Province is 40.41%. Age, annual income, job and sleep duration may affect the development of depressive symptoms among grassroots healthcare workers.

OBJECTIVEWe aimed to construct a recombinant adenovirus Ad-APN-EGFP and to evaluate the transfection efficiency and effect of the adenovirus promoting osteogenesis around implants.

METHODSRecombinant adenovirus Ad-APN-EGFP containing adiponectin (APN) was constructed by DNA sequencing and restriction enzyme digestion. Rat bone marrow stem cells (BMSCs) were transfected in vitro. Transfection efficiency and APN mRNA expression were tested. Animal models of rat femoral epiphysis and hydroxyapatite (HA)-coated implants were established. The Ad-APN-EGFP adenovirus at 10 μL was injected into the defect around HA-coated implants in the experimental group, whereas the same volume of phosphate-buffered saline was injected into the defect in the control group. Osteogenesis around implants was observed by hematoxylin-eosin staining four weeks after implantation.

RESULTSAd-APN-EGFP was successfully constructed by DNA sequencing and restriction enzyme digestion. The transfection efficiency of Ad-APN-EGFP to BMSCs was up to ≥ 90%. APN mRNA expression of BMSCs transfected with Ad-APN-EGFP was higher than that of the control group. Osteogenesis in the experimental group was more evident than that in the control group in vivo.

CONCLUSIONAd-APN-EGFP could be transfected into BMSCs and express APN mRNA at a high level. Ad-APN-EGFP could improve osteogenesis around implants in vivo.

Adenoviridae ; Animals ; Dental Implants ; Durapatite ; Osteogenesis ; Rats ; Transfection

China ; Dental Casting Technique ; Titanium

The cochlea of children with congenital sensorineural hearing loss with normal inner ear structure was measured and analyzed by high-resolution temporal bone CT(HRCT) imaging technique,its application value before cochlear implantation was evaluated and the appropriate electrode was selected.We collected temporal bone HRCT images of 120 patients with congenital sensorineural hearing loss,according to gender divided into two groups,including 60 males and 60 females.We used the PACS software to measure the distance A(the largest distance from the round window to the lateral wall) and the distance H(height of the cochlea) and calculate the cochlear duct length. Reproducibility of these data were evaluated and the results between the different groups were compared.Measurement of parameter values between the intraobserver and interobserver showed great reproducibility. In the male children group,the measured values are shown as distance A［(8.55±0.31)mm］,distance H［(4.57±0.28)mm］and the cochlear duct length(CDL)［(27.59±1.23)mm］; and in the female children group, the measured values are shown as distance A［(8.45±0.32)mm］,distance H［(4.42±0.34)mm］and the cochlear duct length(CDL)［(27.20±1.17)mm.The A,H,and CDL of the male cochlea were greater than those of the female, the difference was statistically significant(<0.05).Measuring the distance A and distance H of the cochlea and calculating the cochlear duct length CDL can be used to select a suitable length of electrode or to customize a personalized electrode. This is a simple and effective assessment method before cochlear implantation..

OBJECTIVETo investigate the reasons for the rarity of metastases in skeletal muscle.

METHODSBy injecting tumor cells (Walker256 rat carcinosarcoma) through the iliac artery (experimental group) and the tail vein (control group), animal models of blood-borne metastases were established. The quadriceps femoris muscle and lungs were observed grossly and microscopically. Immunohistochemistry was applied to investigate the expression of vascular cell adhesion molecule-1 (VCAM-1) in the microvascular endothelium of these organs. Primary culture of rat skeletal muscle cells was established and conditioned medium (MCM) was collected. Effects of MCM on several tumor cell lines and the biochemical characteristics of skeletal muscle delivered tumor factor(s) were tested by MTT assay. Apoptosis and morphological examination were carried out to investigate the antitumor mechanisms of MCM.

RESULTSIn the experimental group, there were no definite metastases observed in muscle cells. In the control group, lung metastases were present in the lungs of all rats that were sacrificed at the 14th day or died spontaneously (17 rats in all). There was no significant difference between the increase in VCAM-1 in quadriceps femoris muscle 7 days after iliac artery injection and that in lungs 7 days after tail vein injection (P > 0.05). In vitro studies showed that the proliferation of tumor cell lines of mouse SP2/0 myeloma, rat Walker256 carcinosarcoma or human chronic granulocytic leukemia K562, human acute lymphatic leukemia HL-60, LS-174-T colon adenocarcinoma, PC3-M prostatic carcinoma and lung giant cell carcinoma with different metastatic potency (PLA801-C with low metastatic potency, PLA801-D with high metastatic potency) was significantly inhibited when cultured with MCM (P < 0.01 - 0.05). Proliferation of malignant cells showed a dose-dependent decrease, to a certain degree. Proliferation of normal rabbit joint epiphysial disk cells (RGP-2) were not affected by MCM. Proliferation of lung giant cell carcinoma cells with high metastatic potency showed a significant decrease even when cultured in highly diluted MCM (6.25% of primary MCM), when compared with the strain of low metastatic potency. Following ultrafiltration, boiling at 100 degrees C, and treatment with trypsin, skeletal muscle delivered tumor factor(s) were found to be a low molecular weight (MW

CONCLUSIONSThe rarity of metastases in skeletal muscles, generally accepted in the clinical setting, can be reproduced in an animal model. It does not seem to be related to VCAM-1 expression in the microvessels of these organs. Skeletal muscle delivered factor(s) play a key role in the mechanism of the rarity of metastases in skeletal muscle.

Animals ; Cell Division ; Humans ; Immunohistochemistry ; Muscle Neoplasms ; pathology ; secondary ; Muscle, Skeletal ; physiology ; Rats ; Rats, Wistar ; Tumor Cells, Cultured ; Vascular Cell Adhesion Molecule-1 ; analysis

Heparinase III (HepIII) is a 73-kDa polysaccharide lyase (PL) that degrades the heparan sulfate (HS) polysaccharides at sulfate-rare regions, which are important co-factors for a vast array of functional distinct proteins including the well-characterized antithrombin and the FGF/FGFR signal transduction system. It functions in cleaving metazoan heparan sulfate (HS) and providing carbon, nitrogen and sulfate sources for host microorganisms. It has long been used to deduce the structure of HS and heparin motifs; however, the structure of its own is unknown. Here we report the crystal structure of the HepIII from Bacteroides thetaiotaomicron at a resolution of 1.6 Å. The overall architecture of HepIII belongs to the (α/α)₅ toroid subclass with an N-terminal toroid-like domain and a C-terminal β-sandwich domain. Analysis of this high-resolution structure allows us to identify a potential HS substrate binding site in a tunnel between the two domains. A tetrasaccharide substrate bound model suggests an elimination mechanism in the HS degradation. Asn260 and His464 neutralize the carboxylic group, whereas Tyr314 serves both as a general base in C-5 proton abstraction, and a general acid in a proton donation to reconstitute the terminal hydroxyl group, respectively. The structure of HepIII and the proposed reaction model provide a molecular basis for its potential practical utilization and the mechanism of its eliminative degradation for HS polysaccarides.
Amino Acid Sequence ; Bacteroides ; enzymology ; Catalytic Domain ; Crystallography, X-Ray ; Heparitin Sulfate ; metabolism ; Kinetics ; Models, Molecular ; Molecular Sequence Data ; Polysaccharide-Lyases ; chemistry ; metabolism ; Substrate Specificity

Objective: To analyze the characteristics and influencing factors of the newly diagnosed cases locally infected with HIV through sexual contact in Jinhua,so as to provide reference for AIDS prevention and control. Methods: An epidemiological survey was conducted among the HIV/AIDS cases diagnosed in 2017 in Jinhua to collect the information about demographic characteristics,local infection and sexual behaviors. The logistic regression model was used to analyze the related factors for local HIV infection. Results: A total of 438 HIV/AIDS cases infected through sexual contact were recruited,with 272（62.10%）cases infected in Jinhua. The proportion of local infection was 86.67%,79.47%,63.04%,69.09%,77.46% and 77.97%,respectively,among those people aged 60 years or over,permanent residents in Jinhua,employees / students,farmers,those who had lived in Jinhua for more than five years,and those who had a HIV testing in one year. The Results of the multivariate logistic regression analysis showed that heterosexual transmission cases who were permanent residents in Jinhua（OR=3.437,95%CI：1.250-9.451）,who had lived in Jinhua for more than five years（OR=3.609,95%CI：1.403-9.284）,who had commercial heterosexual behaviors in Jinhua（OR=5.463,95%CI：2.529-11.803）were more likely to be infected with HIV in Jinhua;homosexual transmission cases who were permanent residents in Jinhua（OR=4.812,95%CI：1.744-13.275）and who had non-commercial,temporary homosexual behaviors in Jinhua（OR=10.641,95%CI：4.369-25.916）were more likely to be infected with HIV in Jinhua. Conclusion Among the HIV/AIDS cases diagnosed in 2017 infected through sexual contact in Jinhua,having permanent residence,long-term residence,commercial heterosexual behaviors and non-commercial,temporary homosexual behaviors were risk factors for local infection.

OBJECTIVETo observe the changes in the activity of dendritic cells (DCs) after carcino-embryonic antigen (CEA) gene transfection mediated by recombinant adeno-associated virus type2 (rAAV) and tumor cell lysate.

METHODSImmature DCs isolated from peripheral blood monocytes of HLA-A11-positive healthy volunteers were infected with the rAAV carrying CEA gene or loaded with tumor cell lysate. The surface markers of the DCs such as CD40, CD 1alpha, and CD86 were analyzed by flow cytometry. Interleukin-12 (IL-12) in the supernatants of DCs and interferon-gamma (IFN-gamma) released by the cytotoxic T lymphocytes (CTLs) were determined by ELISA detection kit. The specific killing activity of CTL against LoVo cells was assessed by MTT assay.

RESULTSThe DCs following antigen loading with the two methods both highly expressed CD40, CD86 and IL-12, and induced specific CTL that specifically recognized and killed LoVo cells, but the killing effect resulting from rAAV infection of the DCs was much better than that induced by tumor cell lysate loading.

CONCLUSIONBoth methods of antigen loading can induce mature DCs from peripheral blood monocyte cells, but rAAV infection of the DCs can be more effective than tumor cells lysate loading. DCs infected with rAAV may have the potential to serve as an adjuvant immunotherapy for patients with colorectal carcinoma.

B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Cancer Vaccines ; biosynthesis ; immunology ; Carcinoembryonic Antigen ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; therapy ; Dendritic Cells ; immunology ; metabolism ; Dependovirus ; genetics ; Genetic Vectors ; Humans ; Interleukin-12 ; metabolism ; Transfection