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

Objective To study the histopathological changes of relevant intemal organs of Macaca mulatta infected with Plasmodium knowlesi ( P. knowlesi). Methods Histopathological examination of 3 monkeys who died of P. knowlesi infection, 2 P. knowlesi infected monkeys who died of treatment failure with artesunate suppository and 1 P. knowlesi infected monkey that was cured by piperaquine phosphate (PQP) but died of trauma and necrosis of the fore limb. Results The heart, liver, spleen, lung, kidney, brain, pancreas, parathyriod, pituitary and lymph nodes showed severe pathological changes in 3 monkeys (No. 1, 7 and 12) who died of P. knowlesi infection and 1 infected monkey (No. 72) who died of treatment failure with artesunate suppository. Red blood cells containing rmalarial parasites and pigments were concentrated in the capillaries of these organs. Malarial pigments were deposited in many organs or phagocytized by macrophages in 1 monkey (No. 131 ), it was cured by piperaquine phosphate but died of trauma and necrosis of the fore limb; cellular atrophy and disappearance of pancreatic islets, parathyroid and pituitary cells were also observed. One monkey (No. 33) treated with artesunate suppository, showed that blood parasites became negative but recrudesced and pituiary later died from a gavage accident. Its organs showed a significant difference to those of the infected monkeys receiving no treatment. Only the liver Kupffer cells and cerebral matrix contained malarial parasites and pigments; many relevant intemal organs showed repair. Conclusion The pathological changes of relevant internal organs of Macaca mulatta infected with P. knowlesi were examined in detail, especially cellular atrophy and the disappearance of pancreatic islets, parathyoid and pituitary cells and myolysis of cardiac muscles. These changes have not previously been reported elsewhere.

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..

Objective To investigate the role of plasma tissue factor (TF) and tissue factor pathway inhibitor-1 (TFPI-1) level and to observe the effect of extrinsic TFPI-1 on no-reflow (NR) in a rabbit model of ischemia/reperfusion. Methods Rabbits were randomized into four groups (n = 10 each): ischemic- reperfusion group (IR, subjected to 120 minutes of coronary artery occlusion and followed by 60 minutes of reperfusion); ischemic- reperfusion TFPI-1 group (100 ng/kg bolus and 1 ng · kg~(-1) · min~(-1) infusion during reperfusion) ; ischemic group (subjected to 180 minutes of coronary artery occlusion) and sham group. The NR area and ischemic area were determined by thioflavin S and Evan's blue staining in vivo. Plasma TF and TFPI-1 levels were measured before operation, before and at 120 minutes post coronary artery ligation, 10 and 60 minutes after reperfusion by ELISA. Results Plasma TF and TFPI-1 levels before and at 120 minutes post coronary artery ligation were similar among the four groups (all P > 0.05). At 10 and 60 minutes after reperfusion, the plasma TF levels in the IR group was significantly higher than those in ischemic group and sham group [10 minutes: (20.7 ±4. 1) pg/ml vs. (13.9 ±2. 2)pg/ml(P <0. 001), (20.7±4. l)pg/ml vs. (13.2±2.6) pg/ml(P<0. 001); 60 minutes; (15.8±2.6) pg/ml vs. (13.5± 1.6) pg/ml(P<0.05), (15.8 ±2.6) pg/ml vs. (12.1 ±0.7) pg/ml (P < 0. 001)] while the plasma TFPI-1 levels were similar among IR, ischemic and sham groups at 10 minutes after reperfusion and at 60 minutes after reperfusion (all P >0. 05). TFPI-1 level [(9.7 ± 1. 6) ng/ml] was significantly lower in the IR group than in the ischemic group [(11.6 ±1.6) ng/ml, P < 0. 05] and sham group [( 10. 1 ±1.3) ng/ml, P < 0. 01] . TF mRNA expression in the NR area in IR group was significantly up-regulated compared to the ischemic group (P<0. 05) and sham group (P <0. 001 ) while TFPI-1 mRNA expression was similar between IR group and ischemic group ( P > 0. 05 ) . NR severity in the ischemic-reperfusion TFPI-1 group was significantly attenuated compared to IR group (0. 39 ±0. 11 vs. 0.54±0.06, P<0.01). Conclusion Upregulated TF mRNA expression in the NR area and increased plasma TF level during reperfusion period, reduced plasma TFPI-1 level during reperfusion period as well as attenuated NR severity by extrinsic application of human rTFPI-1 in this model suggested an important role in the pathogenesis of the NR phenomenon.

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