Autoimmune Diseases ; diagnosis ; immunology ; pathology ; therapy ; Child ; China ; Gastroenterology ; Hepatitis B ; diagnosis ; pathology ; therapy ; Humans ; Infant

BACKGROUND: As a biochemical marker of cytoplasm, the increased activity of creatine kinase (CK) in human spermatozoa is correlated with both the residual cytoplasma and the ratio of sperm with abnormal func tion. It is a marker of mature sperm and associated with the potential of in semination. OBJECTIVE: To investigate the differences of CK activity in sperm and the relative contents of sperm CK-MM and CK-BB isoenzymes between normal fertile males and infertile males, and evaluate its significance in clinical diagnosis of male infertility. DESIGN: Case-control observation. SETTING: Laboratory of Biochemistry, Family Planning Research Insti tute of Sichuan Province. PARTICIPANTS: Ninety-four male infertile patients between January 1999 and October 2000 were selected from the Department of Family Planning Sciences of Sichuan Province, who had no aspermatism with their wives proved to be fertile. The average age of subjects were 31 years. Eighty subjects with the sperm count ＞ 2×1010 L-1 were taken as normal sperm group and 14 subjects with the sperm counts ＜ 2×1010 L-1 were considered as oligospermia group. Semen obtained from 27 healthy males who were normal in routine examinations and with children was taken as the healthy control group.METHODS: Semen sample collected by masturbation after abstinence of 3 to 5 days was incubated at 37 ℃ for liquefication and routinely analyzed.Total activity of CK in sperm was determined by using a kinetic spectrophotometry and the relative contents of CK isozyme was determined by agarose gel electrophoresis followed by density scanning of CK isozyme.MAIN OUTCOME MEASURES: Sperm counts, percentages of viability and motility of sperm, total CK activity and the relative contents of CK-MM and CK-BB isozyme in spermatozoa.RESULTS: A total of 94 enrolled patients and 27 normal controls were involved in the analysis of results. ①Sperm counts, percentage of viability and motility in oligospermia group ( Ⅱ + Ⅲ, WHO method) were obviously lower than those in the healthy control group, and those in the normal sperm group, except the sperm counts, were remarkably lower than the healthy control group [the sperm counts in healthy control group, normal sperm group and oligospermia group was (6.05 ±0.81 )×1010 L-1, (7.76±1.37)×1010 L-1 and (1.46±0.19)×1010 L-1 respectively (P ＜ 0.01). The survival rate in healthy control group, normal sperm group and oligospermia group was 85.1%,56.8% and 58.2%, P ＜ 0.01, and the sperm motility was 62.9% ,34.6% and 29.5% respectively, P ＜ 0.01].②Total activity of sperm CK in oligospermia group was significantly higher than the healthy control group [respectively (9.000±6.117) and (1.933 ±0.943) kat/108 sperm,P＜ 0.05],although that in the normal sperm group (2.800±0.862) kat/108 sperm was a little higher than the healthy control group, while there were no significant difference between the two groups (P ＞ 0.05).③The relative contents of sperm CK-MM isozyme in the normal sperm group and oligospermia group were obviously decreased (that in the healthy control group, normal sperm group and oligospermia group was 30.5%, 19.0% and 18.0% respectively,P ＜ 0.05), which implied a remarkable difference in sperm differentiation between healthy control group and the latter two groups.CONCLUSION: The determination of sperm CK is meaningful for the diagnosis and pathogenesis of oligospermia. More work should be done on the distribution of sperm CK-MM isozyme in different infertile population as well as its importance in the diagnosis of infertility.

Objectives:Using IE 1 gene promoter of Autograph californica nuclear polyhedrosis virus(AcNPV),a transfer vector with an immediately early gene promoter was constructed.　Methods:Transfer vector pAcPIneo which contains neomycin resistance gene(neo)coding sequence downstream of IE 1-promoter was constructed and cotransfected with the wild type of AcNPV DNA into Sf9 insect cells.Recombinant virus was selected by G418 resistance since the neo gene can be expressed in Sf9 cells.　Results:Northern blot hybridzation with 32　P labeled neo gene fragement as probe showed that the neogene was integrated in plogene of AcNPV genome.　Conclusions:Transfer vector with an immediately early gene promoter of baculovirus was constructed successfully,the neo gene was transcribed from the immediately early phase to the very late phase in infected cells.

Objective To evaluate the efficacy of combination therapy with finasteride and doxazosin in benign prostatic hyperplasia/ lower urinary tract symptoms (BPH/LUTS) patients with intravesical prostatic profusion (IPP).Methods A total of 322 BPH/LUTS patients who accepted combination therapy with finasteride and doxazosin were enrolled in this study.Patients were divided into 4 groups according to the degree of IPP:group Ⅰ(IPP＞10 mm),group 2 (IPP between 5 mm and 10 mm),group 3 (IPP＜5 mm),control group (without IPP).All patients were received inasteride 5 mg once per day and doxazosin 4 mg once per day for 6 months.International prostate symptom score (IPSS),prostatic specific antigen(PSA),ultrasonographic and urcdynamic evaluation were compared before and after treatment.The correlations between the above factors and IPP were estimated by Logistic regression analysis.Results After 6 month of treament,the changes of IPP degree and the maximal urinary flow rate (Qmax) had no significant differences in group 1,group 2 and group 3 as compared with before treatment (all P＞0.05).The IPSS in group 1 was not significantly different before and after treatment (P＞0.05).There were significant differences in the PSA level,IPSS,total prostate volume (TPV),transition zone volume (TZV),residual urine volume (PVR) in the 4 groups before and after treatment (all P＜0.05).Logistic regression analysis showed that PVR and Qmax had positive and negative correlations with IPP (P＜0.001 and P=0.024),respectively.Conclusions Combination therapy with finasteride and doxazosin can significantly improve the symptoms of LUST and reduce the total prostate volume in patients with BPH/LUTS,but for BPH patients with IPP,the combination therapy can not effectively alleviate the degree of IPP.The increase of residual urine volume and decrease of Qmax may enhance the risk of bladder outlet obstruction in BPH patients with IPP.

Objective To establish a PCR-RFLP method for detection of CD26 mutation of hemoglobin E(HbE)gene.Methods The genome DNA from the family members who were suspected of having HbE combined with beta-thalassemia was amplified using PCR.The PCR products were digested by restriction endonuclease Mnl I,and then separated on PAGE.The electrophoretical patterns were finally analyzed to confirm if CD26 mutation was present or not.Results Four cases with CD26 mutation in two families were successfully screened out.Conclusion Although the incident frequency of CD26 mutation in Sichuan population is not high,it must not be neglected since it may present alone or combined with other type of thalassemia.PCR-RFLP method described in this study is available in screening mutated HbE gene and treating the disease owing to its simplicity,rapidity and specificity.

Objective To explore the migration of transplanted neural stem cells labeled with SPIO with 4.7T MR system and study cell differentiation by immunohistochemistry in ischemic rats. Methods Rat neural stem cells(NSCs)co-labeled with SPIO mediated by poly-L-lysine and bromode- oxyuridine(BrdU)were transplanted into the middle cerebral artery occlusion(MCAO)rats after 14 days when MCAO model was made successfully.4.7T MR scanner was used to monitor the migration of transplanted cells after 1.2,3,4,5 and 6 weeks post-transplantation.After MRI examination at 6th week,the rats were killed and Prussian blue staining and immunohistochemistric staining were per- formed to study migration and functional differentiation of NSCs.Results Three weeks after trans- plantation,linear hypointensity area derived from migration of labeled NSCs was observed in the corpus callosum adjacent to the injection site.Six weeks after transplantation,linear hypointensity area was moved toward the midline along the corpus callosurn.MRI findings were confirmed by Prussian blue staining and immunohistochemical straining.Co-labeled immunohistochemical methods demonstrated transplanted NSCs could differentiated into astrocytes and neurons.Conclusion MRI is very useful in demonstrating the migration path of labeled NSCs.Transplanted NSCs into ischemic rats can differen- tiated into astrocytes and neurons during the process of migration.

10 mU/L (GroupⅡ,71 cases) when they were one year old.There were 40 healthy children in control group. The genomic DNA from the peripheral blood was extracted and polymerase chain reaction (PCR) and sequencing were employed to detect the IL-12B gene 3′UTR+1188 SNP.Results The frequency of AA,AC and CC genotype in GroupⅠwere 25.7%,44.3% and 30.0% respectively,and 36.6%, 47.9% and 15.5% in GroupⅡ,48.8%,39.0%,12.2%,in control group,respectively.The differ- ence of frequency of CC genotype and non-CC genotype between GroupⅠand GroupⅡwas significant (x~2=17.078,P

Objective To investigate the effect of promoter hypermethylation of O 6 methylguanine DNA methyltransferase (MGMT) gene on colorectal tumorigenesis and progression. Methods The promoter hypermethylation of O 6 methylguanine DNA methyltransferase gene was assayed in 27 sporadic colorectal adenomas, 62 sporadic colorectal carcinomas and 20 normal colorectal mucosa tissues by methylation specific PCR. At the same time, the expression of MGMT protein was studied in the same samples using immunohistochemistry. Results None of the normal colorectal mucosa tissues showed methylated bands. Promoter hypermethylation was detected in 40.7%(11/27) of adenomas and 43.5% (27/62) of carcinomas, respectively. MGMT proteins were expressed in nucleus and cytoplasm of normal colorectal mucosa tissues. Loss of MGMT expression was found in 22.2% (6/27) of adenomas and 45.2% (28/62) of carcinomas, respectively. There were significant difference among them ( P =0.041). Methylation was detected in 5 of the 6 adenomas( P =0.027) and 24 of the 28 carcinomas( P

0.05).The methyla tion rates were higher in tumor tissue than those in normal and adjacent mueosa(P