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2.
Zhonghua Er Ke Za Zhi ; 61(7): 637-641, 2023 Jul 02.
Article in Chinese | MEDLINE | ID: mdl-37385808

ABSTRACT

Objective: To summarize the clinical characteristics and treatments of chronic non-bacterial osteomyelitis with autoimmune hepatitis in children. Methods: A child who had chronic non-bacterial osteomyelitis with autoimmune hepatitis was admitted to the Department of Gastroenterology of the Children's Hospital Capital Institute of Pediatrics at April 2022. The clinical data was retrospectively analyzed. Using the keywords of "chronic non-bacterial osteomyelitis""autoimmune hepatitis" in Chinese and English, the literature from database establishment to December 2022 in CNKI, Wanfang, China Biomedical Literature Database and Pubmed was searched. Combined with this case, the clinical characteristics and treatment of chronic non-bacterial osteomyelitis combined with autoimmune hepatitis were analyzed. Results: A 5 years and 3 months girl was admitted to the Department of Gastroenterology of Children's Hospital, Capital Institute of Pediatrics for "transaminase elevated for 1 year and swelling of right maxillofacial area for half a year". The physical examinations at admission found a 4.0 cm × 4.0 cm swelling area with tenderness before the right ear, abdominal distention with visible abdominal wall vein, firm and enlarged liver (10.0 cm below the xiphoid and 4.5 cm below the right ribs), and splenomegaly (Line Ⅰ 10.0 cm, Line Ⅱ 11.5 cm, and Line Ⅲ 25.0 cm). There was no redness, swelling or restriction of the limbs. Laboratory examination found abnormal liver function with alanine aminotransferase 118 U/L, aspartate aminotransferase 227 U/L, γ-glutamyltransferase 360 U/L, and positive direct anti-human globulin test; immunology test found immunoglobulin G 41.60 g/L and a homogeneous type of antinuclear antibody of 1∶1 000; the autoimmune hepatitis antibody test found a positive anti-smooth muscle antibody (1∶100). Liver biopsy showed moderate interfacial inflammation and the patient was diagnosed with autoimmune hepatitis (International Autoimmune Hepatitis Group 19). The imaging findings showed extensive involvement of the bilateral mandible, while the right side was severe. There were expansile bone changes, thinning of the bone cortex, and significant swelling of the surrounding soft tissue in the mandibular body, mandibular angle, and mandibular ramus. After treatment of glucocorticoid, the swelling of the right maxillofacial region disappeared and the transaminase returned to normal. Only one case was reported before in English and none in Chinese. The two cases were both girls whose main clinical features were joint pain and swelling. The previous case started with pain in both knee joints, and developed liver injury during treatment while this case had liver injury as the initial clinical presentation. Besides, the affected sites and degrees of arthritis in the 2 cases were different. After glucocorticoid treatment, the clinical symptoms were alleviated, and transaminases returned to normal. Conclusions: Chronic non bacterial osteomyelitis may involve the liver and manifest as autoimmune hepatitis. Glucocorticoids therapy is effective.


Subject(s)
Hepatitis, Autoimmune , Osteomyelitis , Female , Humans , Child , Glucocorticoids , Retrospective Studies , Hepatitis, Autoimmune/complications , Hepatitis, Autoimmune/diagnosis , Hepatitis, Autoimmune/drug therapy , Alanine Transaminase , Osteomyelitis/diagnosis , Osteomyelitis/drug therapy
3.
Zhonghua Yi Xue Za Zhi ; 103(20): 1538-1545, 2023 May 30.
Article in Chinese | MEDLINE | ID: mdl-37246003

ABSTRACT

Objective: To analyze high-risk factors affecting BK polyomavirus (BKPyV) infection and to construct a prediction model for BKPyV infection in children after renal transplantation. Methods: The clinical data of 332 children who received allogeneic kidney transplantation in the First Affiliated Hospital of Zhengzhou University from January 2014 to March 2022 were retrospectively collected. According to the BKPyV load level, the dynamic change process of lymphocytes at different time points were analyzed. The factors that have potential influence on BKPyV infection were screened by Cox regression analysis, and the receiver operating characteristic curve (ROC) was used to evaluate the sensitivity and specificity of the predictive model of infection. Results: Among the 332 children, there were 215 males and 117 females; the age of transplantation was (12.2±3.9) years old; 37 cases were preschool (1-5 years old), and 295 cases were post-school age (6-18 years old). BKPyV load in 224 urine samples and 30 blood samples of children were detected. There were 9 cases of BKPyV-associated viruria and 3 cases of BKPyV associated viremia in pre-school children, 76 cases BKPyV associated viruria and 14 cases of BKPyV associated viremia in post-school children. Multivariate Cox regression analysis showed that higher body mass index (BMI) (HR=1.105, 95%CI: 1.020-1.197), antithyroglobulin (ATG) application (HR=2.196, 95%CI: 1.335-3.613), and higher tacrolimus concentration (HR=2.484, 95%CI: 1.298-4.753), higher natural killer (NK) lymphocyte count (HR=1.193, 95%CI: 1.009-1.411), higher CD14++CD16-cell count (HR=1.096, 95%CI: 1.024-1.173) were independent risk factors for BKPyV associated viruria in post-school children. Delayed graft function (DGF) (HR=4.993, 95%CI: 1.555-16.038), Acute rejection (AR) (HR=6.021, 95%CI: 1.930-18.787), higher CD14++CD16-cell count (HR=1.227, 95%CI: 1.081-1.392) were independent risk factors for BKPyV associated viremia in post-school children. The results of ROC curve analysis showed that combined BMI, immune induction drugs, tacrolimus concentration, NK cell count, and CD14++CD16-cell count predicted the occurrence of BKPyV associated viruria in post-school children after kidney transplantation at 0.5, 1, 2, and 5 years with area under curve (AUC) of 0.712 (95%CI: 0.626-0.798), 0.708 (95%CI: 0.612-0.804), 0.754 (95%CI: 0.668-0.840) and 0.767 (95%CI: 0.685-0.849). The sensitivity and specificity of the model were 64.9%, 61.4%, 61.6%, 55.8% and 70.9%, 72.4%, 76.0%, 84.0%, respectively. Combined with DGF, AR, and CD14++CD16-cell counts predicted the occurrence of BKPyV-associated viremia at 0.5, 1, 2, and 5 years after renal transplantation in post-school children with AUC of 0.791 (95%CI: 0.631-0.951), 0.744 (95%CI: 0.547-0.936), 0.786 (95%CI: 0.629-0.946) and 0.812 (95%CI: 0.672-0.948). The sensitivity and specificity of the model were 76.1%, 67.1%, 75.0%, 77.9% and 88.9%, 89.0%, 89.9%, 88.0%, respectively. Conclusions: The postoperative CD14++CD16-cell level can be used as an independent predictor of BKPyV infection in post-school children after renal transplantation. Combined BMI, immune induction drugs, tacrolimus concentration, NK cell count, CD14++CD16-cell count and combined DGF, AR, CD14++CD16-cell count show good fitting effect in predicting the occurrence of BKPyV-associated viruria and viremia after transplantation in post-school children respectively.


Subject(s)
BK Virus , Kidney Diseases , Kidney Transplantation , Polyomavirus Infections , Tumor Virus Infections , Male , Female , Humans , Child, Preschool , Child , Adolescent , Infant , Kidney Transplantation/adverse effects , Retrospective Studies , Tacrolimus , Viremia/etiology , Polyomavirus Infections/epidemiology , Tumor Virus Infections/epidemiology
4.
Zhonghua Yu Fang Yi Xue Za Zhi ; 57(12): 2220-2223, 2023 Dec 06.
Article in Chinese | MEDLINE | ID: mdl-38186179

ABSTRACT

The use of cosmetics in the crowd has the long-term characteristics. The adverse reactions of cosmetics reported in other country in the world suggest that human patch tests and short-term human using test may not be sufficient to evaluate the safety of high-risk new cosmetic raw ingredients, and long-term human using test should be conducted for evaluation. Therefore, this article reviews the key factors that affect long-term human trial trials, such as site of use, single-use amount, frequency of use, duration of use, and subject conditions, providing supportive evidence for standardized safety evaluation standards for long-term human using test of cosmetics.


Subject(s)
Cosmetics , Human Experimentation , Humans
5.
Eur Rev Med Pharmacol Sci ; 24(3): 1041-1053, 2020 02.
Article in English | MEDLINE | ID: mdl-32096174

ABSTRACT

OBJECTIVE: Ovarian cancer is a highly invasive type of cancer. A previous study demonstrated that E-cadherin expression was upregulated in a human ovarian cancer cell line with a high expression of WW domain-containing oxidoreductase (WWOX), which is a tumor suppressor. Also, the migration and invasion ability of these cells was reduced. Snail family members are involved in the epithelial-to-mesenchymal transition (EMT) of ovarian cancer cells, and the expression of Snail family members is regulated by the transcription factor Elf5. The aim of the present research was to elucidate the role of WWOX in EMT of ovarian carcinoma cells through the Elf5/Snail pathway by gain and loss of function approaches in in vitro experiments. MATERIALS AND METHODS: First, a WWOX gene expressing plasmid was transfected into CD133+CD117+ HO8910 ovarian carcinoma cells, and an Elf5 shRNA plasmid was transfected into these cells to assess the changes in EMT-related factors, including Snail1, and the invasive ability of tumor cells ability. Second, the human ovarian carcinoma cell lines HO8910 and SKOV3 were divided into six groups to detect the same indicators. RESULTS: The results demonstrated that the high expression of WWOX resulted in an increased E-cadherin expression, decreased Snail1 activity, and decreased invasion ability in CD133+CD117+ HO8910 cells. Elf5 shRNA transfection did not affect the WWOX expression; however, it decreased the expression of E-cadherin and Elf5 activity, while increasing Snail1 activity and invasion ability in CD133+CD117+ HO8910 cells. It was also observed that WWOX overexpression in HO8910 and SKOV3 cells inhibited the expression of EMT-related proteins and inhibited cell migration and invasion. CONCLUSIONS: Taken together, the results of the present report suggest that WWOX can decrease Snail1 activity by enhancing the activity of Elf5, thus upregulating E-cadherin expression and eventually inhibiting EMT of ovarian carcinoma.


Subject(s)
Carcinoma, Ovarian Epithelial/metabolism , DNA-Binding Proteins/biosynthesis , Epithelial-Mesenchymal Transition/physiology , Ovarian Neoplasms/metabolism , Snail Family Transcription Factors/biosynthesis , Transcription Factors/biosynthesis , Tumor Suppressor Proteins/biosynthesis , WW Domain-Containing Oxidoreductase/biosynthesis , Carcinoma, Ovarian Epithelial/genetics , Cell Line, Tumor , DNA-Binding Proteins/genetics , Female , Gene Expression Regulation, Neoplastic , Humans , Ovarian Neoplasms/genetics , Signal Transduction/physiology , Snail Family Transcription Factors/antagonists & inhibitors , Transcription Factors/genetics , Tumor Suppressor Proteins/genetics , WW Domain-Containing Oxidoreductase/genetics
6.
Zhonghua Er Ke Za Zhi ; 57(2): 150-152, 2019 Feb 02.
Article in Chinese | MEDLINE | ID: mdl-30695893
7.
Sci Rep ; 7(1): 9491, 2017 08 25.
Article in English | MEDLINE | ID: mdl-28842675

ABSTRACT

Grazing is the primary land use in the Hulunber meadow steppe. However, the quantitative effects of grazing on ecosystem carbon dioxide (CO2) fluxes in this zone remain unclear. A controlled experiment was conducted from 2010 to 2014 to study the effects of six stocking rates on CO2 flux, and the results showed that there were significant differences in CO2 fluxes by year, treatment, and month. The effects of light and intermediate grazing remained relatively constant with grazing year, whereas the effects of heavy grazing increased substantially with grazing duration. CO2 flux significantly decreased with increasing grazing intensity and duration, and it was significantly positively correlated with rainfall, soil moisture (SM), the carbon to nitrogen ratio (C/N ratio), soil available phosphorus (SAP), soil NH4+-N, soil NO3-N, aboveground biomass (AGB), coverage, height, and litter and negatively correlated with air temperature, total soil N (TN) and microbial biomass N (MBN). A correspondence analysis showed that the main factors influencing changes in CO2 emissions under grazing were AGB, height, coverage, SM, NH4+-N and NO3-N. Increased rainfall and reduced grazing resulted in greater CO2 emissions. Our study provides important information to improve our understanding of the role of livestock grazing in GHG emissions.

8.
Zhonghua Er Ke Za Zhi ; 54(10): 767-772, 2016 Oct 02.
Article in Chinese | MEDLINE | ID: mdl-27784480

ABSTRACT

Objective: To evaluate the utility of magnetic resonance imaging (MRI) in diagnosis of juvenile dermatomyositis and polymyositis (JDM-PM) in children. Method: Fifty-four patients with JDM-PM in the active stage were enrolled in the study group. Twelve patients with benign acute childhood myositis and forty patients with juvenile idiopathic arthritis (JIA) complicated with myositis were enrolled as controls. MRI imaging of thighs was performed in all patients, fast spin echo T1WI, T2WI, and STIR were obtained in all patients.Muscle biopsy was performed in 41/54 patients with JDM-PM. We compared the value of MRI in diagnosis of JDM-PM with muscle biopsy, electromyography and serum aspartate transaminase (AST), alanine transaminase (ALT), creatine kinase (CK), isoenzyme of creatine kinase (CKMB), lactate dehydrogenase (LDH), hydroxybutyrate dehydrogenase (HBDH) levels. Continuous normally distributed variables were reported as means and continuous non-normally distributed variables as median. Chi-square test and Fisher exact test were used to test differences between MRI and other categorical variables. Result: A total of 54 patients were included. Twenty-seven patients were male and the others were female. Average age of the patients was (7.1±3.5) years (2-13 years); 45(83%) paitests were JDM cases and 9(17%) patients had JPM. All patients had MRI examination. Of the 54 patients, 53 had multiple myositis; 10 out of 50 (19%) patients received second MRI after treatment, 6 out of 10 patients had normal findings, 4 patients showed obviously improved images; 41 out of 54 patients underwent muscle biopsy; 22 out of 41 patients had inflammatory cells infiltration and muscle fiber degeneration. The results of the muscle enzyme tests are as follows: 27 (50%) patients had elevated AST, 24 (44%) patients had elevated ALT, 22 (41%) patients had elevated CK, 18(33%) patients had elevated CKMB, and LDH rose in 30 (56%) patients, HBDH rose in 28(52%) patients. These results suggested that muscle MRI was more sensitive than muscle biopsy and muscle enzyme tests in diagnosis of JDM-PM. Conclusion: Patients with JDM-PM showed diffuse patchy hyperintense signals on T2WI of their thighs. MRI may be a sensitive, reliable, and noninvasive tool for clinical diagnosis and theraputic evaluation of JDM-PM.


Subject(s)
Dermatomyositis/pathology , Myositis/pathology , Polymyositis/pathology , Adolescent , Alanine Transaminase , Biopsy , Chi-Square Distribution , Child , Child, Preschool , Creatine Kinase , Electromyography , Female , Humans , Magnetic Resonance Imaging , Male
9.
Zhonghua Er Ke Za Zhi ; 54(6): 446-50, 2016 Jun 02.
Article in Chinese | MEDLINE | ID: mdl-27256233

ABSTRACT

OBJECTIVE: To investigate the value of brain magnetic resonance imaging (MRI) in evaluating the intracranial injuries in patients with juvenile onset systemic lupus erythematosus (SLE). METHOD: Data of brain MRI, CT, electroencephalogram (EEG), cerebrospinal fluids analysis and clinical features of the central nervous system of 44 patients from March 2007 to March 2015 with juvenile onset SLE who were not treated with glucocorticoids (Gcs) and immunosuppressive agents (Is) were retrospectively analyzed and compared. RESULT: Twenty-seven out of 44 patients demonstrated abnormal signs on brain MRI, including encephalatrophy, cerebral infarction, demyelination, encephalorrhagia, vertebral arteriostenosis and abnormal signals on the brain diffusion-weighted imaging (DWI). Sixteen patients had clinical features of the central nervous system involvement, fifteen had continuous headache, nine had continuous dizziness, seven had convulsions, three had hemiplegia, one had blurred vision. Physical examination of the nervous system: ten patients had abnormal signs, all had cervical rigidity, five showed pyramidal sign, three showed loss of muscle tone, two with cranial neuropathies and one had paresthesia. EEG: Ten patients showed abnormal waves on EEG, all showed diffused slow-waves, and five showed sharp waves and spikes. Cerebrospinal fluids analysis: six patients had abnormal results, five of them had cell count elevation and one had cell count and protein elevation, while there was glucose and chloride degression. Brain CT: Eight patients received CT scan, two showed cerebral infarction. χ(2) test was used to compare the differences among head MRI, EEG, cerebrospinal fluid analysis, physical examination of the nervous system, clinical features of the nervous system, the difference was significant(χ(2)=12.055, P=0.001; χ(2)=19.627, P=0.001; χ(2)=3.859, P=0.049; χ(2)=12.055, P=0.001). CONCLUSION: Brain MRI may be a better method in early diagnosis of intracranial injuries than CT, EEG, cerebrospinal fluid analysis and physical examination of the nervous system. Patients with juvenile onset SLE should receive brain MRI after diagnosis in order to investigate the intracranial injuries. Abnormal signals on the DWI are the signs of active disease.


Subject(s)
Brain/diagnostic imaging , Lupus Vasculitis, Central Nervous System/diagnostic imaging , Cerebral Infarction , Cranial Nerve Diseases , Diffusion Magnetic Resonance Imaging , Early Diagnosis , Electroencephalography , Headache , Humans , Retrospective Studies , Seizures
10.
Lupus ; 19(10): 1195-203, 2010 Sep.
Article in English | MEDLINE | ID: mdl-20504833

ABSTRACT

Glomerular microthrombosis (GMT) is a common vascular change in patients with lupus nephritis (LN). The mechanism underlying GMT is still unknown. In our previous study, we found that the level of IgG anti-beta2 glycoprotein I (beta2GPI) antibodies was higher in the LN-GMT group than in the LN-non-GMT group, which indicated that anti-beta2GPI antibodies may play a role in GMT formation. Many studies have demonstrated that the activation of the classical complement pathway may play a critical role in fetal loss and aPL-induced thrombosis formation. To investigate whether complement activation plays a role in GMT formation and to evaluate its relationship with aPL, we prospectively investigated deposition of C4d in 155 renal biopsy specimens of LN patients. The results revealed a strong relationship between the intensity of glomerular C4d staining and the presence of microthrombi (p < 0.001). The detection rate of IgG anti-beta2GPI antibodies was higher in the LN-GMT group than in the LN-non-GMT group (p < 0.05). Further, the intensity of glomerular C4d staining was significantly related with IgG anti-beta2GPI antibodies (p < 0.05). The results of our study suggest that anti-beta2GPI antibodies may play a role in GMT formation, and this process might involve complement activation.


Subject(s)
Complement C4b/metabolism , Lupus Nephritis/complications , Peptide Fragments/metabolism , Thrombosis/physiopathology , beta 2-Glycoprotein I/immunology , Adult , Autoantibodies/immunology , Biopsy , Female , Humans , Immunoglobulin G/immunology , Kidney Glomerulus/immunology , Kidney Glomerulus/pathology , Lupus Nephritis/immunology , Male , Middle Aged , Prospective Studies , Thrombosis/etiology , Thrombosis/immunology , Young Adult
11.
Arch Androl ; 48(5): 369-78, 2002.
Article in English | MEDLINE | ID: mdl-12230823

ABSTRACT

The functional relationship between GABA(A) and GABA(B) receptors in regulating acrosome reaction (AR) of rat spermatozoa was demonstrated by studying the differential effects of a GABA(B) agonist and an antagonist on the process. AR rates were determined using the chlortetracycline staining assay. The induction of AR in rat sperm by GABA was found to be a biphasic phenomenon; i.e., AR rates increased with increasing GABA concentrations up to <5 micro M and at higher concentrations of the neurotransmitter (>5 micro M), there was a reductionin the AR rates. This biphasic phenomenon is apparently due to the differential interaction of the neurotransmitter with GABA receptor subtypes in a dose-dependent manner; i.e., GABA(A) receptors (stimulatory) are primarily activated at low concentration of GABA, while GABA(B) receptors (inhibitory) become activated at higher concentrations. This hypothesis is supported by the present findings that treatment with saclofen, a GABA(B) receptor antagonist, did not influence the AR rates effected by GABA at low concentrations; while the AR rates were maintained at the maximum level at higher concentrations of GABA, resulting in the elimination of the biphasic phenomenon. Baclofen, a GABA(B) receptor agonist, blocks the AR activating action of GABA at both low and high concentrations. It would appear that the induction of AR in rat sperm by GABA is regulated by the proportionality of activated GABA(A) and GABA(B) receptors acting as a yin-yang control.


Subject(s)
Acrosome Reaction/physiology , Acrosome/physiology , Baclofen/analogs & derivatives , Receptors, GABA-A/physiology , Receptors, GABA-B/physiology , gamma-Aminobutyric Acid/physiology , Animals , Baclofen/pharmacology , GABA Agonists/pharmacology , GABA Antagonists/pharmacology , In Vitro Techniques , Male , Progesterone/pharmacology , Rats , Rats, Wistar
12.
Biochem Biophys Res Commun ; 283(1): 243-7, 2001 Apr 27.
Article in English | MEDLINE | ID: mdl-11322795

ABSTRACT

gamma-Aminobutyric acid (GABA) can mimic and potentiate the action of progesterone in initiating the acrosome reaction (AR) of mammalian sperm, indicating that sperm contain receptors for GABA. This contention was validated by identifying the receptor (R) subtype, GABA(A)R, in mammalian sperm. In the present study a second subtype, GABA(B)R, was identified in rat testis and sperm. Total RNAs of rat testis and sperm were prepared and used as template to synthesize the respective cDNAs by the RT-PCR method. Two splice variants of the cDNA coding GABA(B)R1 (GABA(B)R1a and GABA(B)R1c) and GABA(B)R2 were identified. Extracts of rat testis, spermatogenic cells and sperm contained two proteins with estimated molecular sizes of 130 and 100 kDa, corresponding to GABA(B)R1a and GABA(B)R1c/lb, respectively, determined by Western blot using polyclonal anti-GABA(B)R1 antibody. By an indirect immunofluorescence technique, GABA(B)R1 was located on the head of rat sperm. The present finding is the first direct demonstration that mammalian sperm contain GABA(B)R.


Subject(s)
Receptors, GABA-B/metabolism , Spermatozoa/metabolism , Testis/metabolism , Alternative Splicing , Animals , Blotting, Western , DNA, Complementary/genetics , Fluorescent Antibody Technique , Male , Molecular Weight , Protein Isoforms/genetics , Protein Isoforms/metabolism , RNA, Messenger/metabolism , Rats , Receptors, GABA-B/genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Spermatozoa/cytology , Testis/cytology
14.
Arch Androl ; 45(3): 169-80, 2000.
Article in English | MEDLINE | ID: mdl-11111865

ABSTRACT

The HSD-I gene codes a human sperm membrane protein (hSMP-1) and has been assigned the accession number U12978. The gene is located on human chromosome 9, region p12-p13. When the 1.7-kb cDNA of HSD-I was digested sequentially with EcoRI, BamHI, and HindIII, a 550-bp cDNA fragment was formed, which codes for the extracellular domain. This fragment was cloned into the asd+ vector pYA3149 to construct pYA3149R. The recombinant plasmid was used to transform an avirulent deltacva, deltacrp, deltaasd vaccine strain of Salmonella typhimurium chi4550. The hSMP-1 component was localized on the surface of the head of mature rat spermatozoa by an immunofluorescence technique using polyclonal anti-hSMP-1 antibodies. Since rat sperm contain hSMP-1, this rodent can be used to assay the immunogenicity of pYA3149R. Female Wistar rats were immunized by oral administration of the recombinant Salmonella. Anti-hSMP-1 antibodies in blood and vaginal washes of immunized animals were determined. Both body fluids contained significant amounts of the antibodies, showing that the recombinant Salmonella is an effective oral immunogen in rats.


Subject(s)
Membrane Proteins/immunology , Salmonella typhimurium/immunology , Spermatozoa/immunology , Vaccines, Attenuated , Vaccines, Synthetic , Administration, Oral , Animals , Antibody Formation , Antigens/immunology , Antigens, Surface , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunoglobulin A/analysis , Male , Membrane Proteins/analysis , Membrane Proteins/genetics , Molecular Sequence Data , Rats , Rats, Wistar , Restriction Mapping , Salmonella typhimurium/genetics , Spermatozoa/cytology , Vagina/immunology
15.
Arch Androl ; 45(3): 239-46, 2000.
Article in English | MEDLINE | ID: mdl-11111873

ABSTRACT

Serum was obtained from an infertile woman having antibodies with sperm agglutinating activity. The antibodies interacted with a human sperm membrane protein (hSMP-1) with an estimated Mr of 55 kD. The gene (HSD-1) coding hSMP-1 was isolated from a human testis cDNA expression library and assigned the accession number U12978. The cDNA was conjugated to a prokaryotic expression vector to construct the recombinant vector, pRSET-HSD-I, which was expressed in Escherichia coli. The recombinant hSMP-1 was isolated and used to immunize rabbits to raise polyclonal antibodies. Usingan immunocytochemical technique, hSMP-1 protein was immunolocalized in germ cells of human testis at all stages of spermatogenesis. mRNAs were prepared from 16 different human tissues and analyzed by Northern blot using HSD-1 as probe. A positive reaction was elicited only with testis mRNA. The present findings suggest that the expression of hSMP-1 gene is testis-specific and occurs during the early stages of germ cell differentiation. In a comparative study, the location of the hSMP-I protein in sperm and in germ cells of the seminiferous tubules of rats was determined. The target antigen was immunolocated on the head and tail of rat sperm and in late spermatids and spermatozoa of rat testis. These results suggest that, in the rat, the HSD-1 gene is expressed during spermiogenesis.


Subject(s)
Gene Expression Regulation, Developmental , Germ Cells/cytology , Membrane Proteins/genetics , Spermatozoa/cytology , Spermatozoa/physiology , Animals , Antigens, Surface , Cell Differentiation , Female , Humans , Immunohistochemistry , Male , Membrane Proteins/analysis , Molecular Sequence Data , Rabbits , Rats , Testis/cytology , Testis/physiology
16.
Sheng Wu Gong Cheng Xue Bao ; 16(1): 42-5, 2000 Jan.
Article in Chinese | MEDLINE | ID: mdl-10883274

ABSTRACT

Plasmid pRL-B1 was constructed from detoxifying gene(called B1) of pesticide resistant Culex and from plasmid pRL-439 containing the strong promoter PpsbA. E. coli-cyanobacteria shuttle expression plasmid pDC-B1 was constructed from shuttle vector pDC-8 and from recombinant plasmid pRL-B1, then it was transferred into Synechococcus sp. PCC7942 by triparental conjugative transfer. The existence of B1 was detected by Southern analysis, and the expression of B1 was confirmed by enzyme activity analysis of detoxification of transgenic cyanobacteria. Experimental results indicated that the transgenic cyanobacteria could degrade beta-naphthyl acetate(beta-NA), a specific substrate of esterase. The enzyme activity of transgenic strain was higher than that of the wild type. It may be the first report on transformation of detoxify gene of pesticide resistant culex into Synechococcus strain.


Subject(s)
Culex/genetics , Cyanobacteria/genetics , Genes, Insect , Pesticides/metabolism , Animals , Cloning, Molecular , Drug Resistance , Plasmids
17.
Cell Res ; 10(1): 51-8, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10765983

ABSTRACT

Some recent studies indicated that GABAergic system is involved in mammalian sperm acrosome reaction (AR), but direct evidence pertaining to the expression of gat1 in mammalian sperm is not yet demonstrated. In this study, we evaluated the presence of 67kDa GAT1 protein and mRNA in rat testis by Western blotting and reverse transcription-polymerase chain reaction. Meanwhile, immunohistochemical and immunofluorescent analyses also identified GAT1 protein on the elongated spermatid and sperm. These results indicated that rat testis is a novel site of gat1 expression. Further studies should be taken to explore the role of GAT1 protein on sperm acrosome reaction.


Subject(s)
Carrier Proteins/genetics , Carrier Proteins/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolism , Membrane Transport Proteins , Organic Anion Transporters , Spermatozoa/metabolism , Animals , GABA Plasma Membrane Transport Proteins , Immunohistochemistry , Male , RNA, Messenger/metabolism , Rats , Reverse Transcriptase Polymerase Chain Reaction , Testis/metabolism , gamma-Aminobutyric Acid/metabolism
18.
Biochem Biophys Res Commun ; 270(2): 581-7, 2000 Apr 13.
Article in English | MEDLINE | ID: mdl-10753667

ABSTRACT

We have previously reported an association of 14-3-3epsilon isoform with calmodulin. Using the voltage-clamp technique, the present study investigated the potential role of 14-3-3 in modulating the Ca(2+)-activated Cl(-) channel (CaCC) endogenously expressed in Xenopus oocytes. Injection of 14-3-3epsilon antisense oligodeoxynucleotides resulted in potentiation of the ionomycin-induced Cl(-) current, while 14-3-3 peptide and calmodulin inhibitor, W13, suppressed the antisense-potentiated current. The data suggest that 14-3-3epsilon plays an inhibitory role in modulating the CaCC by interacting with the calmodulin-dependent pathway. The potential role of 14-3-3epsilon in other tissues and its therapeutic potential for cystic fibrosis are discussed.


Subject(s)
Calcium/metabolism , Chloride Channels/metabolism , Proteins/physiology , Tyrosine 3-Monooxygenase , 14-3-3 Proteins , 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid/pharmacology , Animals , Base Sequence , Calcium-Calmodulin-Dependent Protein Kinase Type 2 , Calcium-Calmodulin-Dependent Protein Kinases/metabolism , Chloride Channel Agonists , Chloride Channels/antagonists & inhibitors , Humans , Ionomycin/pharmacology , Oligonucleotides, Antisense/genetics , Oligonucleotides, Antisense/pharmacology , Proteins/genetics , Xenopus
19.
Cell Res ; 9(3): 201-8, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10520602

ABSTRACT

Primordial germ cells (PGCs), as precursors of mammalian germ lineage, have been gaining more attention as a new resource of pluripotent stem cells, which bring a great possibility to study developmental events of germ cell in vitro and at animal level. EG4 cells derived from 10.5 days post coitum (dpc) PGCs of 129/svJ strain mouse were established and maintained in an undifferentiated state. With an attempt to study the differentiation capability of EG4 cells with a reporter protein: green fluorescence protein, and the possible application of EG4 cells in the research of germ cell development, we have generated several EG4-GFP cell lines expressing enhanced green fluorescence protein (EGFP) and still maintaining typical characteristics of pluripotent stem cells. Then, the differentiation of EG4-GFP cells in vitro as well as their developmental fate in chimeric embryos which were produced by aggregating EG4-GFP cells to 8-cell stage embryos were studied. The results showed that EG4 cells carrying green fluorescence have a potential use in the research of germ cell development and other related studies.


Subject(s)
Cell Lineage , Germ Cells/metabolism , Luminescent Proteins/metabolism , Animals , Chimera , Fibroblasts/cytology , Fibroblasts/metabolism , Green Fluorescent Proteins , Intestines/embryology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred ICR , Mice, Inbred Strains , Mice, Transgenic , Plasmids/metabolism , Transfection
20.
J Med Invest ; 46(1-2): 19-28, 1999 Feb.
Article in English | MEDLINE | ID: mdl-10408153

ABSTRACT

Antisperm antibodies can cause infertility by interacting with spermatozoa through immunoglobulin binding protein thereby blocking their penetrance of cervical mucus and/or by interfering with sperm-egg interaction. However, these antibodies appear not to be cytotoxic to embryos since a high implantation rate and consequently high pregnancy rate were achieved by IVF-ET treatment of women with antisperm antibodies. Also the finding that these antibodies do not appear to cause any deleterious clinical symptoms and have yet be associated with infertility suggested that sperm antigens are promising candidates in the development of immunocontraceptives. Some synthetic peptides corresponding to segments of human sperm antigens have effectively induced infertility in female rats when administered as an immunogen. Different peptides, adjuvants and routes of administration should be studied to determine the optimum conditions for inducing high antisperm antibody titers in the host. Moreover, identification of various steps and factors that are involved in regulating the production of antisperm antibodies such as immunoglobulin binding factor may open new paths in the treatment of immunological infertility and at the same time lead to a more effective immunocontraceptive.


Subject(s)
Antibodies/immunology , Infertility, Female/immunology , Spermatozoa/immunology , Animals , Female , Fertilization in Vitro , Humans , Infertility, Female/therapy , Male , Pregnancy , Rats , Sperm-Ovum Interactions/immunology
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