OBJECTIVETo prepare monoclonal antibodies of mice anti-human sperm protein 22 (SP22) and to identify their specificities.

METHODSBALB/c mice were immunized with human SP22, monoclonal antibodies were prepared by hybridoma technique and the sensitivity and specificity of SP22 McAb were investigated by ELISA and Western-blot assay, respectively. The distribution of human SP22 in sperm were shown by immunohistochemistry.

RESULTSThree strains of hybridoma cells were obtained, with the affinity constant (K) of 1.0 x 10(7) L/mol and the titers were 1:10(3) and 1:3,200 in the mixed supernatant of cell cultures and abdominal dropsy, respectively. IgG isotype of the antibody was identified as IgG1. Western blot demonstrated that there was a specific recognition between human SP22 and the obtained monoclonal antibody. Immunohistochemistry displayed that human SP22 mainly distributed on the acrosome surface of human sperm.

CONCLUSIONThe monoclonal antibodies of anti-human 22 was prepared by the technique of hybridoma cell has higher titer and specificity, which can combine specially with the SP22 protein on the surface of human sperm.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; Blotting, Western ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Hybridomas ; secretion ; Immunohistochemistry ; Male ; Mice ; Mice, Inbred BALB C ; Microtubule-Associated Proteins ; immunology ; metabolism ; Spermatozoa ; metabolism

Multifunctional sperm protein 22 (SP22) is expressed ubiquitously and related to quite a few diseases. Located on the sperm surface, SP22 has an enzymatic activity that may assist sperm in penetrating into the ovum. SP22 may be carcinogenic in conspiracy with the factor ras. Among all species SP22 is highly conservative, which demonstrates its importance to life. More and more studies indicate that SP22 is directly correlated with male infertility and Parkinsons disease. This article summarizes recent researches on SP22 in the gene structure, protein structure and functional characteristics.
Animals ; Gene Components ; Humans ; Male ; Microtubule-Associated Proteins ; chemistry ; genetics ; physiology ; Protein Conformation ; Protein Deglycase DJ-1 ; Rats ; Spermatozoa ; physiology

OBJECTIVETo investigate the epidemiological features of the pathogens responsible for prostatitis in the Changshu area, and offer some evidence for the clinical treatment of prostatitis.

METHODSThis study included 2 306 cases of prostatitis that were all clinically confirmed and subjected to pathogenic examinations in 3 hospitals of Changshu area from 2008 to 2012. Neisseria gonorrhoeae, mycoplasma urealyticum and chlamydia trachomatis were detected by nucleic acid amplification ABI 7500, the bacterial data analyzed by VITEK-2 Compact, the drug-resistance to antibacterial agents determined using the WHONET 5.6 software, and the enumeration data processed by chi-square test and curvilinear regression analysis using SPSS 19.0.

RESULTSThe main pathogens responsible for prostatitis were found to be Staphylococcus haemolyticus (30%), Staphylococcus epidermidis (12%), Enterococcus faecalis (9%), Escherichia coli (6%), Staphylococcus warneri and Staphylococcus aureus (3%), Mycoplasma urealyticum (8%), chlamydia trachomatis (5%) and Neisseria gonorrhoeae (6%). Statistically significant increases were observed in the detection rates of Escherichia coli (chi2 = 17.56, P<0.05), Mycoplasma urealyticum (chi2 = 8.73, P<0.05), Chlamydia trachomatis (chi2 = 8.73, P<0.05) and Enterococcus (chi2 = 8.22, P<0.05), but not in other pathogens. The resistance rates of Gram-positive bacteria to erythromycin and benzylpenicillin G were both above 45%, but with no significant difference between the two, those of Oxacillin (chi2 = 10.06, P<0.05) and Cefoxitin (chi2 = 9.89, P<0.05) were markedly increased, but those of quinolones, gentamycin and clindamycin remained low, except rifampicin (chi2 = 11.09, P<0.05). The resistance rates of Gram-negative bacteria to cefazolin and ampicillin were relatively high (mean 57.3%), and those to ceftriaxone (chi2 = 11.26, P<0.05) and trimethoprim sulfamethoxazole (chi2 =11.00, P< 0.05) significantly high; those to amikacin, cefepime, piperacillin/tazobactam and imipenem remained at low levels with no significant changes. However, the resistance rates of mycoplasma urealyticum to ciprofloxacin (chi2 = 11.18, P<0.05) and azithromycin (chi2 = 9.89, P<0.05) were remarkably increased.

CONCLUSIONGram-positive bacteria are the major pathogens responsible for prostatitis, but Escherichia coli, enterococcus and sexually transmitted disease pathogens are found to be involved in recent years. Quinolones and aminoglycosides are generally accepted as the main agents for the treatment of Gram-positive bacterial infection. However, rational medication for prostatitis should be based on the results of pathogen isolation and drug sensitivity tests in a specific area.

Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Bacterial ; Gram-Negative Bacteria ; drug effects ; Gram-Positive Bacteria ; drug effects ; Humans ; Male ; Prostatitis ; epidemiology ; microbiology

ObjectiveTo investigate the distribution of the human papilloma virus (HPV) and its genotypes in the male outpatients at the clinics of sexually transmitted diseases (STD) in Changshu and analyze its association with the primary clinical symptoms so as to provide some evidence for the prevention and treatment of HPV infection in men.

METHODSWe collected exfoliated cell samples from the external genitals of 602 male outpatients at the STD clinics in Changshu from February 2016 to February 2018, extracted and amplified nucleic acids from the samples, and detected the HPV genotypes using the gene chip technique. We performed statistical analyses on the types of symptoms in clinical diagnosis and their correlation with the genotypes of HPV using the chi-square test.

RESULTSThe HPV positive rate in the male STD clinics was 48.2%, of which 47.2 % fell into the low-risk type, 30.0% with multiple infections. The main genotypes included HPV types 6, 11, 39, and 52, and the main HPV-related clinical symptoms were verruca (43.1%) and erythra (41.0%). Low-risk types 6 and 11 accounted for a significantly higher percentage than the high-risk types in the verruca patients (60.0% vs 15.0%, , P < 0.05), but showed no statistically significant difference from the latter in the erythra patients (38.7% vs 38.7%, P > 0.05). The incidence of low-risk infection was remarkably higher than that of high-risk infection in the acrobystitis and balanitis patients (P < 0.05), while the high-risk types constituted a markedly higher percentage than the low-risk and high- and low-risk mixed types in the asymptomatic men at physical examination (84.6% vs 0.0% and 15.4%, P < 0.05).

CONCLUSIONSThe HPV positive rate was as high as 48.2% in the males at the STD clinics in Changshu, and the main infection type was low-risk genotype single infection. The clinical symptoms of low-risk infection were mainly verruca and prepuce balanitis, and the high-risk type was mostly asymptomatic at physical examination.

OBJECTIVE: To explore the mechanism of catgut embedding at back- points on nonalcoholic steatohepatitis (NASH) in rats based on IKK/IKB/NF-κB signaling pathway and downstream inflammatory factors. METHODS: Eighty SPF SD rats were selected, among them 10 rats were selected divided into a normal group (group A), and the remaining 70 rats were fed with high-fat diet to establish NASH model. At the end of 12 weeks, 10 rats were randomly selected to verify whether the model establishment was successful. Then the remaining 60 rats were randomly divided into a model group (group B), a catgut embedding at back- points group (group C), a catgut embedding at abdominal points group (group D), an acupuncture at back- points group (group E), a sham catgut embedding group (group F) and a western medication group (group G), 10 rats in each group. The rats in the group C were treated with catgut embedding at "Ganshu" (BL 18), "Pishu" (BL 20), "Weishu" (BL 21) and "Shenshu" (BL 23); the rats in the group D were treated with catgut embedding at "Daheng" (SP 15), "Fujie" (SP 14), "Huaroumen" (ST 24) and "Tianshu" (ST 25); the rats in the group E were treated with acupuncture at the same acupoints as the group C; the rats in the group F were treated with catgut embedding at back- points but the needle did not enter subcutaneous tissue gamma; the rats in the group G were treated with intragastric administration of vitamin E capsule. All the treatment was given for 4 weeks. The rats in the group A were fed with normal diet until the end of 16 weeks without any intervention. The rats in the group B continued to be fed with high-fat diet until the end of 16 weeks. After the intervention, the liver index was calculated; the liver histomorphology was observed by HE staining; the liver function [alanine aminotransferase (ALT), gamma glutamyl transferase (γ-GGT), alkaline phosphatase (ALP)] and blood lipid [serum total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL)] were measured by serum biochemistry. The serum levels of TNF-α, IL-6 and IL-1βwere detected by ELISA, and the expressions of IKK-α, NF-κBp65, IL-6, IL-1β and TNF-α proteins in liver tissue were detected by Western blot. The temperature of the conception vessel and the governor vessel was measured by infrared thermography. RESULTS: Compared with the group A, the obvious steatosis and inflammatory cell infiltration were observed in the group B, and the body weight, liver wet-weight and liver index were all increased (<0.01). Compared with the group B, the liver tissue morphology in the group C, the group D, the group E and the group G was improved in varying degrees, and the liver index was decreased (<0.05), which was the most significant in the group C (<0.05). Compared with the group A, the ALT, γ-GGT, ALP, TG, TC, LDL, TNF-α, IL-6 and IL-1β were all increased in the group B (<0.01); compared with the group B, the ALT, γ-GGT, ALP, TG, TC, LDL, TNF-α, IL-6 and IL-1β in all intervention groups were all decreased in varying degrees (<0.01, <0.05), which was the most significant in the group C (<0.01). Compare with the group A, the expressions of IKK-α, NF-κBp65, TNF-α, IL-6 and IL-1βproteins in the group B were all increased (<0.01); compared with the group B, the expressions of IKK-α, NF-κBp65, TNF-α, IL-6 and IL-1βproteins in all intervention groups were decreased in varying degrees (<0.05), which was the most significant in the group C (<0.01). Compared with the group A, the temperature of the conception vessel and governor vessel was decreased in the group B (<0.01). Compared with the group B, the temperature of the conception vessel and governor vessel was all increased in the group C, the group D and the group E (<0.01); the temperature of the conception vessel in the group C was similar to that in the group D (>0.05), while the temperature of the governor vessel in the group C was superior to that in the group D (<0.05). CONCLUSION The catgut embedding at back- points might inhibit the activation of IKK/IKB/NF-κB signaling pathway to interrupt the inflammatory cascade, and reduce the "second hit" of inflammatory factors on liver, which could slow down NASH progress and prevent and treat NASH.
Acupuncture Points ; Animals ; Catgut ; NF-kappa B ; Non-alcoholic Fatty Liver Disease ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

OBJECTIVETo explore the effect and mechanism of norcantharidin (NCTD) on hematopoiesis function in leucopenia model rat induced by cyclophosphamide (CTX).

METHODSLeucopenia model was replicated in SD rat with cyclophosphamide(CTX) and model animal was treated with NCTD. Peripheral blood and bone marrow tissue samples were collected from the rats in each experimental group. Peripheral white blood cells (WBC) were counted and analyzed by automatic blood cell analyzer. Histopathologic changes of the biopsied bone marrow tissues were observed by histopathological techniques. The cell cycle and apoptosis rate of bone marrow cells were detected by flow cytometry. Immunohistochemical method was applied to observe the expression of apoptosis-related proteins BCL-2 and BAX in bone marrow.

RESULTSAfter NCTD treatment in model rats, the WBC count of peripheral blood obviously increased, the cell structure of bone tissue significantly recovered, NCTD could promote the cell proliferation and cycle changes of bone marrow cells, inhibit the bone marrow cell apoptosis and necrosis induced with CTX, up-regulate the expression of apoptosis-related protein BCL-2 and downregulated the BAX.

CONCLUSIONNCTD can stimulate the bone marrow hematopoiesis and promote recovery of peripheral white blood cell level in the leukopenia model induced by CTX, and its mechanism may be related with NCTD regulating bone marrow cell cycle and with NCTD inhibiting cell apoptosis.

Animals ; Apoptosis ; Bone Marrow ; Bone Marrow Cells ; Bridged Bicyclo Compounds, Heterocyclic ; Cell Cycle ; Cell Proliferation ; Cyclophosphamide ; Disease Models, Animal ; Flow Cytometry ; Hematologic Diseases ; Hematopoiesis ; Rats ; Rats, Sprague-Dawley

Objective: To understand the basic characteristics of previously reported patients with hepatitis C and analyze the related factors affecting their antiviral treatment. Methods: A convenient sampling method was adopted. Patients who had been previously diagnosed with hepatitis C in the Wenshan Prefecture of Yunnan Province and Xuzhou City of Jiangsu Province were contacted by telephone for an interview study. The Andersen health service utilization behavior model and related literature were used to design the research framework for antiviral treatment in previously reported hepatitis C patients. A step-by-step multivariate regression analysis was used in previously reported hepatitis C patients treated with antiviral therapy. Results: A total of 483 hepatitis C patients, aged 51.73 ± 12.06 years, were investigated. The proportion of male, agricultural occupants who were registered permanent residents, farmers and migrant workers was 65.24%, 67.49%, and 58.18%, respectively. Han ethnicity (70.81%), married (77.02%), and junior high school and below educational level (82.61%) were the main ones. Multivariate logistic regression analysis results showed that married patients with hepatitis C (OR = 3.19, 95% CI: 1.93-5.25, compared with unmarried, divorced, and widowed patients) with high school education or above (OR = 2.54, 95% CI: 1.54-4.20, compared with patients with junior high school education or below) were more likely to receive antiviral treatment in the predisposition module. Patients with severe self-perceived hepatitis C in the need factor module (compared with patients with mild self-perceived disease, OR = 3.36, 95% CI: 2.09-5.40) were more likely to receive treatment. In the competency module, the family's per capita monthly income was more than 1,000 yuan (compared with patients with per capita monthly income below 1,000 yuan, OR = 1.59, 95% CI: 1.02-2.47), and the patients had a high level of awareness of hepatitis C knowledge (compared with patients with a low level of knowledge, OR = 1.54, 95% CI: 1.01-2.35), and the family members who knew the patient's infection status (compared with patients with an unknown infection status, OR = 4.59, 95% CI: 2.24-9.39) were more likely to receive antiviral treatment. Conclusion: Different income, educational, and marital statuses are related to antiviral treatment behavior in hepatitis C patients. Family support of hepatitis C patients receiving hepatitis C-related knowledge and their families knowing the infection status is more important in promoting the antiviral treatment of patients, suggesting that in the future, we should further strengthen the hepatitis C knowledge of hepatitis C patients, especially the family support of hepatitis C patients' families in treatment.
Humans ; Male ; Antiviral Agents/therapeutic use* ; China ; Hepatitis C/drug therapy* ; Hepacivirus ; Logistic Models