Effect of experimental varicocele on structure and function of epididymis in adolescent rats

<abstract>Aim: To study the effect of experimental left varicocele (ELV) on epididymal structure and function in adolescent Sprague-Dawley rats. Methods: ELV was induced by partial ligation of the left renal vein. Sham-operated animals served as the controls. Four and 8 weeks after the operation, the histological, ultrastructural and biochemical (alpha-glucosidase activity and carnitine content) changes in different segments of the epididymis were observed. Results: In the treated animals, there were degeneration of the epididymal epithelium and edema of the interstitial tissue; numerous shedding cells, residual bodies, deformed sperm and macrophages appeared in the epididymal lumen. Morphometric measurement indicated a significant reduction in the epididymal tubular diameter (P<0.05) and a significant increase in the epididymal interstitial area (P<0.05) compared with the controls. Ultrastructural study showed sparse microvilli of the columnar epithelium, increased and enlarged lysosomes in the principal cells with defected organelles and the presence of large cytoplasmic vacuoles. The protein and carnitine contents and the alpha-glucosidase activity in the caput, corpus and cauda epididymis of the ELV rats were lower than those of the controls (P< 0.05). Conclusion: There were structural and functional changes in the epididymis of adolescent ELV rats, which may contribute to the infertility caused by varicocele.

Quantitative (stereological) study of incomplete spermatogenic suppression induced by testosterone undecanoate injection in rats

Aim: To evaluate the key lesions in spermatogenesis suppressed partially by testosterone undecanoate (TU) treatment. Methods: Adult male SD rats were treated with vehicle or TU (19 mg/kg) injection (i.m.) every 15 days for 130 days. The numbers of all types of cells (nuclei) in the seminiferous tubules and the interstitial tissue were estimated using a contemporary stereological tool, the optical disector. Results: In response to TU treatment, the numbers of non-type B spermatogonia, type B spermatogonia and late elongated spermatids per testis were reduced to 51 %, 66 % and 14 % of the controls, respectively. The conversion ratios from type B spermatogonia to early spermatocytes and pachytene spermatocytes were not significantly affected and the ratios to the later germ cell types fell to 51 % - 65 % of the controls. Less than 1.0 % of immature round spermatids were seen sloughing into the tubule lumen, 4.0 % of elongated spermatids retained in the seminiferous epithelium, and about half of the elongated spermatid nuclei appreciably malformed. Leydig cells were atrophied but their number and the peritubular myoid cell number per testis were unchanged. Conclusion: Double inhibition of spermatogenesis (i.e. inhibition at spermiation and spermatogonial conversion to type B spermatogonia), a scenario seen in the monkey and human following gona-dotrophin withdrawal, was not sufficiently effective for a complete spermatogenic suppression in the rat after TU treatment, probably due to ineffective inhibition of the Leydig cell population and therefore the intra-testicular testosterone levels.

Reversible effect of testosterone undecanoate injection on spermatogenesis in rats

Aim: To study the effect of testosterone undecanoate (TU) injection on spermatogenesis in rats. Methods:Twenty adult SD rats received vehicle or TU (8 mg/kg, 19 mg/kg or 625 mg/kg) injection, im, every 15 days for 60days, and another 38 animals received similar treatments for 130 days with half of them undergoing a recovery phase of120 days (5 rats for each treatment). At the end of the treatment, testes were removed and the diameter of the seminif-erous tubules and the number of late elongated spermatids (steps 15-19) per testis were estimated with stereologicalmethods as a measure of the spermatogenic efficiency. Results: Low dose (8 mg/kg) TU treatment virtually hadno effect on spermatogenesis. A dose of 19 mg/kg slightly suppressed spermatogenesis 60 days after treatment, and se-vere suppression occurred after another 70 days of dosing. Spermatogenesis was completely recovered at the end of therecovery phase. Large dose (625 mg/kg) TU treatment did not significantly affect spermatogenesis and was well toler-ated by animals. Conclusion: TU injection reversibly suppresses spermatogenesis in rats.

Analysis of Change in Sperm Quality of Chinese FertileMen druing 1981～1996

By literature search, 114 papers for fertile m ale sperm quality including 256 set data w ere collected from 9 292 personsand 11 726 assaysinvolving 39 citiesand coun- ties. Results of analysis show ed a decrease in m ean concentration of sperm from 103.02×106 /m l (1983) to 83.84×106/m l (1996), sperm m otility w as decreased from 75.11(1982) to 67.27(1996) and percentage of sperm w ith norm alm or- phology w asreduced from 85.02(1983) to 77.89(1996), thusshow ing thedefi- nite negative correlation and being statistically significant (P< 0.05). Total sperm countsw ere decreased from 355.34×106(1984) to 262.26×106(1996) and the m ean sem inalvolum ew asdecreased from 3.31 m l(1981) to 2.97 m l(1996), both tending to decline butnotbeing statistically significant(P> 0.05). Itisinteresting to notethat although Chinese sperm quality is better, itdeclines significantly faster than thatof w estern countriesatthesam eperiod. Itispossible thatthe decline of sperm quality is due to problem of environm entalquality. Theauthorssuggesttheemphasisof basicre- search in relevantfields.

Expression and localization of VCX/Y proteins and their possible involvement in regulation of ribosome assembly during spermatogenesis

Variable Charge X/Y (VCX/Y) is a human testis-specific gene family that localized on X and Y chromosomes. In this study, VCY protein was expressed in E. coli in the form of glutathione-S-transferase (GST) fusion protein. With the purified fusion protein as antigen, the anti-GST-VCY antibody was generated and the localization of VCY protein in human testis was determined by immunohistochemistry. In the testis seminiferous epithelium. VCY proteins were highly expressed in nuclei of germ cells. Using propidium iodide staining and green fluorescent protein (GFP) tag technologies, VCY and VCX-8r proteins were mainly localized in the uucleoli of COST cells. In addition, the colocalization for VCY and VCX-8r in COS7 cells was also observed. With VCY cDNA as bait, a cDNA fragment of acidic ribosomal protein PO was obtained using yeast two-hybrid system. All the information above indicates that VCX/Y protein family might be involved in the regulation of ribosome assembly (hiring spermatogenesis.

Long-term Safety Observations on Side Effects and Com -plications of Non-surgical Sterilization by Chem ical In-stillation of the Fallopian Tubal

The present objective is to observe the side effects and complications of fem ale tubalsterilization by phenol-atabrinepaste (PAP) and phenolm ucilage (PM). 1 705 eligible w om en w ere divided random ly into tw o groups, 871 usinig PAP and 834 w om en using PM. Results: Thefeverratesw ere8.0and 4.4( P< 0.01 ) respec- tively in group PAP and group PM. Infection rates of the appendages w ere 2.3 and 1.2 respectively. The incidence of chem ical peritonitis, w ithin the lim its of pelvic cavity, w ere 1.1and 2.4respectively. Perforation of uterus, pelvic ab- scessand ectopicpregnancy w ere notfound during thisobservation. Slightabdom inal pain after the proceduresw ere 33.6and 30.8respectively. Slightvaginalbleed- ing after the procedure w as 23.2 and 18.0 respectively. Menstruation of the w om en w as not affected by the instillation. This five-year follow -up study neither show ed a positivePap testnor a potentialdeseasecaused by thesterilizing agents. This technique isa safe and efficientm ethod for fem ale sterilization.

Identification of differentially expressed genes of primary spermatocyte against round spermatid isolated from human testis using the laser capture microdissection technique

The method of laser capture microdissection (LCM) combined with suppressive subtractive hybridization (SSH) was developed to isolate specific germ cells from human testis sections and to identify the genes expressed during differentiation and development. In the present study, over 10,000 primary spermatocytes and round spermatid cells were successfully isolated by LCM. Using the cDNAs from primary spermatocytes and round spermatids, SSH cDNAs library of primary spermatocyte-specific was constructed. The average insert size of the cDNA isolated from 75 randomly picked white clones was 500 bp, ranging from 250 bp to 1.7 kb. Using the dot-blot method, a total of 421 clones were examined, resulting in the identification of 390 positive clones emitting strong signals. Partial sequence of cDNAs prepared from each clone was determined with an overall success rate of 84.4%. Genes encoding cytochrome c oxidase II and the rescue factor-humanin were most frequently expressed in primary spermatocytes, suggesting their roles involved in meiosis.

Similarity between Spatial-temporal Expression Patterning of HOXAll and Progesterone Receptor in Human Endometrium

Objective To study HOXAll expression and its correlation with that of the progesterone receptor (PR) gene in human endometriumMethods In situ hybridization and semi-quantitative RT-PCR were used. Results HOXAll mRNA was detected in both stromal and glandular cells of normal endometrium by in situ hybridization. But the expression levels in the glandular cells had a dramatic decline or even disappearance at mid-secretory stage. Semi-quantitative RT-PCR analysis, however, demonstrated that the total expression levels of HOXAll mRNA were markedly increased in the mid-secretory endometrium, which suggested that there was an increased expression in stromal cells. Similar results were obtained for PR gene expression in human endometrium by in situ hybridization and semi-quantitative RT-PCR analysis.Conclusion HOXAll gene spatial and temporal expression patterns were similar to that of PR gene in endometrium across menstrual cycle, and HOXAll was closely related to the endometrial proliferation and differentiation during menstrual cycle, especially the establishment of receptive status in implantation.

The Effect of Proteins From Defined Stages of Seminiferous Tubules on Secretory Function of Sertoli Cell in Rats

The influence of the proteins secreted by defined stages of seminiferous tubules of rat on the production of total protein and transferrin secreted by Sertoli cell of 10 days' rat was investigated in the bicameral chamber system. The results indicated that the proteins secreted by stage Ⅱ-Ⅵ Ⅶ-Ⅷ,Ⅸ-Ⅻ，ⅩⅢ-Ⅰ of seminiferous tubules stimulated the production of total protein secreted by Sertoli ceils by an increase of 290±117%, 253±59%. 268±27%, 290±25% (X±SD) respectively. The autoradiography either from electrophoresis of(^35S)-methionine labeled total protein or transferrin seereted by Sertoli cells exhibited the stimulatory, effect from defined stage of serainiferous tubules. Most de novo proteins synthesized by Sertoli cells were secreted into the apical chamber. However, no significant differenee in stimulatory effeet from different stages of seminiferous tubules has been observed.

Preliminary Identification of Human Nonserum Oviduct Specific Proteins by Using Electrophoresis and Immunoblotting Analysis

The present paper reported the preliminary results of identification of humannonserum oviduct specific proteins. The 1D-SDS-polyacrylamide gel electrophoresis (PAGE) and 2D-SDS-PAFE in conjunction with the immunoblotting assay were used in the present study. The results showed that the nonserum oviduct specific proteins with MW130, 100 and 80 kD existed in human oviduct fluid or oviductal extract. In addition, the antibody against pig oviduct antigens could more strongly cross-react with human oviduct antigens, mainly recognizing 130,116 and 100 kD proteins from human oviduct. It is suggested that in human oviduct there are some specific antigens possessing some similar epitopes to those in pig oviducts. This result seems to be consistent with predominant cross reactivity existing in antigens of porcine and human zona pellucida.

Quantitative and qualitative changes in serum luteinizing hormone after injectable testosterone undecanoate treatment in hypogonadal men

Aim: To clarify the immuno-active LH (i-LH) and bioactive LH (b-LH) responses and qualitative changes in the cir-culating LH to testosterone undecanoate (TU) injection. Methods: Eight men with Klinefelter's syndrome were re-cruited for the study. They received crossover injections of TU at doses of 500 and 1000 mg. Serum i-LH and b-LHlevels before and at various time intervals after TU injection were measured and the serum i-LH, b-LH, b-LH/i-LH(B/I) and testosterone/sex hormone-binding globulin (T/SHBG) ratio in LH-responders and LH non-responders werecompared. Results: A parallel suppression of serum i-LH and b-LH was consistent with their overall high correlationbetween each other ( r = 0.84, P < 0. 001). Mean serum i-FSH levels were decreased by TU injection at both doseswithout dose-response effects. LH-responders had lower baseline serum i-LH and b-LH, and higher E_2 levels and T/SHBG ratio. There was a quantitative change in serum LH as induced by TU without qualitative change within LH-re-sponders os LH-non-responders. Conclusion: A high loading dose (1000 mg) of TU is important for the initial sup-pression of LH. With the lower dose (500 mg), repeated injections will be required to attain such LH suppression forthe purpose of fertility regulation. The lower baseline serum i-LH level may be an intrinsic characteristic of LH-respon-ders.

NATURAL CONCEPTIVE PROBABILITY IN NORMAI CHINESE MARRIED WOMEN

The purpose of this study is related with not only the fertility of newly married couple but also the time required for conception after marriage or cessation of contraception and their influential factors. A total number of 1974 couples were investigated from January

Characterization of rtSHSp13 gene encoding a development protein involved in vesicular traffic in spermiogenesis

A cDNA, designated as rtSH3p13, was isolated from a rat testis cDNA library. It consists of 1463 bp nuclear acids,which encodes a protein of 312 amino acids and was assigned the GenBank accession number AF227439. The deduced rtSH3p13 protein is a truncated isoform of SH3p13 as a result of mRNA alternative splicing. It is mainly expressed in the rat testis, detected in spermatids at the steps 8-19 of spermiogenesis, and found around the acrosome. During postnatal development, rtSH3p13 appears on day 18 and reaches maximum on day 60. Further experimental results suggested that rtSH3p13 forms a complex with activated epidermal growth factor receptor (EGFR) and interacts with synaptojanin I. Surprisingly, similar to SH3 domain, the V region of rtSH3p13 also inhibits endocytosis in CHO cells.Our results reveal a link between an rtSH3p13-synaptojanin-clathrin complex-mediated formation of pits and the process of spermiogenesis.

A Five-year Follow-up Study on the Efficacy and Safety of Female Tubal Sterilization by Chemical Instillation

A multi center randomized clinical trial was adopted in the present study. 1705 eligible women with identical demographic and gynecological characteristics were divided into two groups: 871 using phenol atabrine paste (PAP) and 834 using phenol mucilage (PM). The five year follow up rates for the two groups were 97.5% and 98.1%, respectively. The multiple decrement life table analysis showed that there was a significant difference in the gross cumulative failure rates between the two groups. The 60th month gross cumulative failure rates for PAP group and PM group were 4.61% and 11.87%, respectively, thus indicating the efficacy of PAP to be significantly higher than that of PM. 97.7% users had cervical smear examinations, but no suspected cancer cells or cancer cells were found. Meanwhile, there was no known diseases related to the chemical reagent. This follow up study indicates that tubal sterilization by chemical instillation is a simple,safe and efficient female method. It suggested that this method would be introduced to a wider use on the basis of modification in the composition of chemical agents, standardized operational procedure, and improved administrative regulation in the use of this technology.

Randomized Multi-center Study of Baofuxin for Treatment of Bleeding Side-effect Induced by IUD

To observe the efficacy of Baofuxin for treatment of bleeding side effect induced by IUD. Method The study is a multi-center trial. The subjects were randomly allocated into two groups, Baofuxin group (90 cases) and Indomethacin group (90 cases). In the Baofuxin group, the subjects took the medicine on the first day of menses, once a bag, twice a day for 10 days. In the Indomethacin group,only one capsule was taken once a time, twice a day for 7 days. The treatment was given for three menstrual cycles. The subjects were asked to record their bleeding/spotting by using menstrual diary card not only during the treatment cycles but also during the three months previous and after the treatment cycles. The menstrual profile was analyzed by using MDSv2. 3 program that was provided by WHO. Results Within each 90-day reference period of treatment and post-treatment cycles, the number of bleeding/spotting days decreased obviously and bleeding/spotting free days were greatly increased. Both medicines have little effect on number of episode of bleeding/spotting. The subjects who thought the treatment were highly effective were 81. 1% in the Baofuxin group and 56.2% in the Indomethacin group respectively (P <0.01). Conclusion Both Bat,fuxin and indomethacin are highly effective on treatment of bleeding side effect induced by IUD, but Baofuxin had longer effects and was more acceptable.

Regulation of C－Fos mRNA Expression in Sertoli Cells by cAMP，Ca^（＋＋），and protein Kinase C－mediated Pathways

The role of second messenger pathways, cyclic AMP, calcium,and protein and protein kinase C(PKC) in the transcriptional regulation of c-fos proto-oncogene expression in rat Sertoli cells was investigated, c-fos expression wasmonitored by Northern blot analysis.Although the action of FSH on Sertolicells is considered to be mediated by cAMP, dibutyryl cAMP(db cAMP), apotent membrane permeable analog of cAMP,induced much less c-fos mRNA expression than FSH(<50%), suggesting that additional cAMPindependent mechanisms may mediate the effect of FSH on c-fos. Specificintracellular inhibitors of PKC decreased c-fos induction in response to FSHby more than 50%. Ionomycin, which increases intracellular free calciumconcentration, induced c-fos expression significantly. These datademonstrate that Sertoli cell c-fos mRNA expression in under multifactorial regulation by cAMP, calcium, and PKC.

Advances of Contraception in China

Family planning is an important component of reproductive health. Reduction in total fertility rate is closely related to the decrease of maternal and infant mortality rate and the advances in maternal and child health care. In the last two decades the reduction of total fertility rate from 5.6% in the 70s to 2. 5% in average at present occurs along with the decrease in maternal and infant mortality. This achievement should be attributed partly to the success in the development of

An Epidemiologic Survey of Infertility in Dalian District of China

An epidemiologic survey of infertility was carried out in Dalian district from September to December 1989. A two-stage random sampling was used to select subjects. Date were collected by household, interview with standardized questionnaire.Quality control was strictly executed. The respondent rate was 96%. A total of 5 918 couples were included in the analysis. The prevalence of primary infertility was 1.01% in this district. There was no significant difference between urban and