BACKGROUNDVolatile anesthetics (VAs) may affect varied and complex physiology processes by manipulating Ca(2+)-calmodulin (CaM). However, the detailed mechanism about the action of VAs on CaM has not been elucidated. This study was undertaken to examine the effects of VAs on the conformational change, hydrophobic site, and downstream signaling pathway of CaM, to explore the possible mechanism of anesthetic action of VAs.

METHODSReal-time second-harmonic generation (SHG) was performed to monitor the conformational change of CaM in the presence of VAs, each plus 100 µmol/L Ca(2+). A hydrophobic fluorescence indicator, 8-anilinonaphthalene-1-sulfonate (ANS), was utilized to define whether the VAs would interact with CaM at the hydrophobic site or not. High-performance liquid chromatography (HPLC) was carried out to analyze the activity of CaM-dependent phosphodiesterase (PDE1) in the presence of VAs. The VAs studied were ether, enflurane, isoflurane, and sevoflurane, with their aqueous concentrations 7.6, 9.5, 11.4 mmol/L; 0.42, 0.52, 0.62 mmol/L; 0.25, 0.31, 0.37 mmol/L and 0.47, 0.59, 0.71 mmol/L respectively, each were equivalent to their 0.8, 1.0 and 1.2 concentration for 50% of maximal effect (EC50) for general anesthesia.

RESULTSThe second-harmonic radiation of CaM in the presence of Ca(2+) was largely inhibited by the VAs. The fluorescence intensity of ANS, generated by binding of Ca(2+) to CaM, was reversed by the VAs. HPLC results also showed that AMP, the product of the hydrolysis of cAMP by CaM-dependent PDE1, was reduced by the VAs.

CONCLUSIONSOur findings demonstrate that the above VAs interact with the hydrophobic core of Ca(2+)-CaM and the interaction results in the inhibition of the conformational change and activity of CaM. This in vitro study may provide us insight into the possible mechanism of anesthetic action of VAs in vivo.

Adenosine Monophosphate ; analysis ; Anesthetics, Inhalation ; pharmacology ; Anilino Naphthalenesulfonates ; Calmodulin ; antagonists & inhibitors ; chemistry ; physiology ; Cyclic Nucleotide Phosphodiesterases, Type 1 ; analysis ; Fluorescence ; Humans ; Hydrophobic and Hydrophilic Interactions

The GATA family proteins are a group of zinc finger transcription factors that are expressed in human and mammalian animals and play an important role in mammalian organ morphogenesis, cell proliferation and sex differentiation. GATA-4 and GATA-6 have been identified in the ovaries and testes of humans, mice, pigs and chickens. GATA-4 contributes to fetal male gonadal development by regulating the genes that mediate Müllerian duct regression and the onset of testosterone production. GATA-4 and GATA-6 are localized in and regulate the function of the ovarian and testicular somatic cells of fetal mice, especially granulosa cells, thecal cells, Sertoli cells and Leydig cells. GATA-4 is also present in the germ cells of fetal and prepubertal mice.
Animals ; Chickens ; Female ; GATA4 Transcription Factor ; metabolism ; GATA6 Transcription Factor ; metabolism ; Humans ; Male ; Mice ; Ovary ; embryology ; Reproduction ; Swine ; Testis ; embryology ; Transcription Factors ; classification

OBJECTIVETo explore the characteristics and distribution of GATA-4 in the testis of male mice.

METHODSParaffin sections were obtained from the testes of 24 male B6SJLF1/J mice, aged 0 day (n = 6), 2 weeks (n = 6), 4 weeks (n = 6) and 6 weeks (n = 6), and the expressions of GATA-4 in the testis were observed by the immunohistochemical ABC method and DAB visualization at different times.

RESULTSPositive expressions of GATA4 were found in the Sertoli cells and Leydig cells of all the mice, but significantly higher in the 4- and 6-week-old than in the 0-day and 2-week-old groups (P < 0.01). And they were also observed in the germ cells of the 4- and 6-week-old mice, significantly higher in the latter than in the former (P < 0.01).

CONCLUSIONGATA-4 exists in the testis of male mice, which has provided a morphological base for sex determination and differentiation and hormone regulation in the testis.

Animals ; Cell Differentiation ; GATA4 Transcription Factor ; metabolism ; Germ Cells ; metabolism ; Leydig Cells ; metabolism ; Male ; Mice ; Sertoli Cells ; metabolism ; Testis ; cytology ; metabolism

OBJECTIVETo investigate the effects of experimental left varicocele (ELV) on the vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase-1 (Flt-1) proteins in the testis and epididymis of adolescent rats, and to find out the correlation of the two proteins with varicocele-induced male infertility.

METHODSWe established the ELV model in adolescent male SD rats, and detected the expressions of VEGF and Flt-1 proteins in the testis and epididymis by immunohistochemistry at 2 and 4 weeks after surgery.

RESULTSCell- and region-specific expressions of VEGF and Flt-1 were observed in the testis and epididymis of the ELV and control groups. Statistical analysis showed that, in comparison with the corresponding control groups, the 2- and 4-week ELV groups exhibited a notable increase in the VEGF protein expression in the hibateral testis and epididymis (P < 0.01, P < 0.05); the Flt-1 expression was obviously upregulated in the hibateral testis and epididymis of the 2-week ELV group (P < 0.01, P < 0.01), but remarkably reduced in the hibateral testis and left epididymis of the 4-week ELV group (P < 0.01, P < 0.05), with no statistic difference in the right epididymis (P > 0.05).

CONCLUSIONELV can cause changes in the expressions of VEGF and Flt-1 proteins in the testis and epididymis of adolescent rats, and consequently affect spermatogenesis and spermiotelcosis, which may be one of the causes of varicocele-induced male infertility or subfertility.

Animals ; Disease Models, Animal ; Epididymis ; metabolism ; Gene Expression ; Male ; Rats ; Rats, Sprague-Dawley ; Testis ; metabolism ; Varicocele ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism

OBJECTIVETo study the expressions of the vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase-1 (Flt-1) in the testis, epididymis and epididymal sperm of adolescent rats and explore the functions of both the proteins in the male reproductive system.

METHODSThe expressions of VEGF and Flt-1 were detected in 20 adolescent SD rats, immunohistochemical staining used for both the testis and the epididymis and immunofluorescent staining for sperm.

RESULTSVEGF and Flt-1 proteins were specifically present in the testis, epididymis and sperm. In the testis, VEGF immunoreactive particles were localized in the cytoplasm of spermatogenic cells, the developing acrosome of spermatids, Sertoli cells and Leydig cells, while Flt-1 expressed mainly in the developing acrosome of spermatids and Leydig cells. In the epididymis, the cell-specific and region-specific expressions of VEGF and Flt-1 proteins were observed in the principal cells of epididymal epithelia, VEGF in the whole epididymis, while Flt-1 only in the caput and cauda segments. Both VEGF and Flt-1 were localized in the acrosome of the sperm head as well as in the neck, middle and principal segments of the sperm tail.

CONCLUSIONThe specific expression patterns of VEGF and Flt-1 in the rat testis, epididymis and sperm indicate that they may independently or collectively affect spermatogenesis and spermiotelcosis in either an autocrinological or a

Animals ; Epididymis ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Sexual Maturation ; Spermatozoa ; metabolism ; Testis ; metabolism ; Vascular Endothelial Growth Factor A ; biosynthesis ; Vascular Endothelial Growth Factor Receptor-1 ; biosynthesis

Objective To evaluate the efficacy and safety of short-term treatment including fluoroquinolones anti-tuberculosis drugs for rifampicin resistant pulmonary tuberculosis(TB)in those areas carrying out the 'TB control project'.Methods TB cases involved in this study were from TB drug resistance surveillance in Heilongiiang province,Zhejiang province and Shenzhen city from 2004 to 2006.TB cases with rifampicin resistant were randomly divided into the treatment group(including fluoroquinolones anti-tuberculosis drugs group)and the control group(re-treatment regimen group).The treatment group was treated wim 3RFT AM ofx Pto PAS-INH/5RFT ofx Pto PAS.INH while the control group was treated with 3 H3R323E3S3/5 H3R3E3.Efficacy of short-term treatment was analyzed by per-protocol analysis(PP analysis)and intention-to-treat analysis(ITT analysis)while drug adverse reactions was also observed.Results (1)154 patients with rifampicin resistant pulmonary tuberculosis were recruited among them,25(16.2%)were only resistant to rifampicin,114(74.0%)to MDR-TB and 15(9.8%)to others(resistant R+S,resistant R+E and resistant R+E+S).114 TB cases completed the fuIl course of treatment,with 71 in the treatment group and 43 in the control group.(2)Sputum negative conversion rate of the treatment group and the control group were 78.9%and 65.1%(X2CMH=4.558,P=0.011)respectively,by per-protocol analysis.Sputum negative conversion rate of the treatment group and the control group were 65.9%and 40.6%(X2CMH=0.272,P=0.001)respectively,by intention-to-treat analysis.The sputum negative conversion rate of the treatment group was higher than in the control group when treating rifampicm resistant pulmonary tuberculosis and MDR-TB patients.(3)Three patients withdrew in each of the two groups because ofadverse effects to the drugs.Rates of adverse reaction to drugs appeared to be 23.9%(17/71)and 18.6%(8/43)in the treatment and in the control groups,with no statistically significant difference between the two groups.Conclusion The efficacy of treatment including fluoroquinolones anti-tuberculosis drugs group seemed beaer than the re-treatment regimen group in treating patients with rifampicin resistant pulmonary tuberculosis and those MDR-TB patients.