Quality Evaluation of Branches of Juglans mandshurica by HPLC-DAD Fingerprint Combined with HPLC-Q-TOF-MS/MS Characterization of Constituents / 中国实验方剂学杂志

Objective:To establish a high performance liquid chromatography （HPLC） fingerprint of branches of <italic>Juglans mandshurica</italic> and to evaluate the quality of the samples from different producing areas and in different harvest periods. Method:Chromatographic separation was performed on an Agilent Poroshell 120 SB-C₁₈ column （2.1 mm×100 mm， 2.7 μm） for gradient elution with mobile phase of 0.2% formic acid solution （A）-0.2% formic acid acetonitrile solution （B） （0-5 min， 5%-10%B； 5-25 min， 10%-16%B； 25-40 min， 16%-22%B； 40-45 min， 22%-45%B； 45-50 min， 45%-65%B； 50-52 min， 65%-100%B； 52-55 min， 100%B） at a flow rate of 0.3 mL·min^-1. The column temperature was 30 ℃ and the detection wavelength was 270 nm. The quality of branches of <italic>Juylans mandshurica</italic> was evaluated by similarity evaluation， cluster analysis， principal component analysis and partial least squares-discriminant analysis. The chemical constituents of the samples were identified by HPLC coupled with quadrupole time-of-flight mass spectrometry （HPLC-Q-TOF-MS/MS）. The mass spectrometry was conducted in negative ion mode with electrospray ionization（ESI）. Data were acquired over a range of <italic>m</italic>/<italic>z</italic> 100-1 700 for MS and <italic>m</italic>/<italic>z</italic> 50-1 700 for MS/MS. Result:A total of 19 common peaks were confirmed in 40 batches of samples， and the similarity ranged from 0.430 to 0.995， of which the similarity of samples collected in spring and winter seasons （April， May and December） was greater than 0.90， while the similarity of most samples collected in summer （July to September） was low. Multivariate statistical analysis showed that the samples were divided into two groups according to the harvest time， but there was no obvious classification rule for the samples from different producing areas. The contents of most constituents in the samples collected in spring and winter were higher than those collected in summer. The result illustrated that different harvest periods had great influence on the quality of branches of <italic>J</italic>.<italic> mandshurica</italic>. Compared with the samples collected in summer， the quality of samples collected in spring and winter was better. A total of 22 peaks were proved to be the main constituents that contributed to the difference between samples collected in different seasons. A total of 83 chemical components were identified by HPLC-Q-TOF-MS/MS， including 49 tannins， 7 organic acids， 14 naphthalene derivatives， 1 flavonoid， 6 anthracene derivatives， 2 lignans， 3 diarylheptanoids and 1 saccharide. Totally 13 common peaks were identified. Of the peaks that contributed to discriminate samples collected in different season， 19 peaks were identified and most of them were tannins. Conclusion:The established HPLC fingerprint can provide useful information for the quality evaluation of branches of <italic>J</italic>.<italic> mandshurica</italic>. Tannin is the main constituents in the samples. Harvest period has great influence on the quality of branches of <italic>J</italic>.<italic> mandshurica</italic>.

Gujing Maisiha Tablets combined with natural vitamin E for the treatment of idiopathic asthenospermia / 中华男科学杂志

Objective@#To investigate the therapeutic efficacy and safety of Gujing Maisiha Tablets combined with natural vitamin E in the treatment of idiopathic asthenospermia.@*METHODS@#This study included 135 outpatients with idiopathic asthenospermia received in our hospital from February 2015 to January 2016. We randomly divided them into a treatment group (n = 65, aged 22－44 ［mean 32.8］ yr) and a control group (n = 55, aged 21－43 ［mean 33.7］ yr) to be treated with Gujing Maisiha Tablets combined with natural vitamin E or natural vitamin E only, both for 90 days. We obtained total sperm motility and the percentage of progressively motile sperm (PMS) from the patients before and after medication and evaluated the clinical effects by comparing the collected parameters and pregnancy rates between the two groups.@*RESULTS@#The baseline total sperm motility and PMS were (25.23 ± 5.57)% and (17.53 ± 5.78)% in the treatment group, with no statistically significant differences from (26.05 ± 6.77)% and (15.11 ± 6.55)% in the control (P >0.05). After 90 days of medication, both the treatment and the control groups showed remarkable increases in total sperm motility (［48.73 ± 8.66］% and ［36.54 ± 8.09］%, P <0.05) and PMS (［32.77 ± 6.04］% and ［26.99 ± 6.87］%, P <0.05). However, both total sperm motility and PMS were significantly higher in the treatment than in the control group after medication (P <0.05), and so was the total rate of clinical effectiveness (73.85% vs 54.55%, P <0.05). No adverse reactions were observed in either of the two groups during the treatment.@*CONCLUSIONS@#Gujing Maisiha Tablets combined with natural vitamin E is safe and effective for the treatment of idiopathic asthenospermia.

Microbial community in a full-scale drinking water biosand filter.

To remove turbidity and minimize microbiological risks, rapid sand filtration is one of main drinking water treatment processes in the world. However, after a long-term operation, sand particles will be colonized by microorganisms which can remove biodegradable organic matters and nitrogen compounds. In this study, 16S rRNA gene clone library analysis was applied to characterize the microbial community in a full-scale biosand filter used for drinking water treatment. The results indicate that phylum Nitrospirae and class Alphaproteobacteria were the dominant bacterial groups in the biosand sample collected from the upper filter layer. The dominance of Sphingomonas species might pose a microbiological risk. This work could provide some new insights into microbial community in drinking water biofilter.

One-hole high ligation of internal spermatic vein by embryonic natural orifice transumbilical laparoscopy: a recommendable treatment of varicocele / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the one-hole method for high ligation of the internal spermatic vein by embryonic natural orifice transumbilical laparoscopy.</p><p><b>METHODS</b>We used the one-hole method for high ligation of the internal spermatic vein by embryonic natural orifice transumbilical laparoscopy in the treatment of 15 cases of varicocele, 13 in the left and 2 in the right side, and appraised the treatment results by follow-up 1 month after the surgery.</p><p><b>RESULTS</b>All the operations succeeded and no complications developed. The average operation time was 28 minutes and the mean hospital stay was 4 days. Symptoms were significantly relieved in all the patients and the scars were inconspicuous at follow-up.</p><p><b>CONCLUSION</b>The one-hole method is a novel option for high ligation of the internal spermatic vein by embryonic natural orifice transumbilical laparoscopy in the treatment of varicocele. It is recommendable for its advantages of simple procedure, less pain, few complications, quick recovery, mini-invasiveness and cosmetic acceptability.</p>

Testis homotransplantation: a report of 12 cases / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of testis homotransplantation in the treatment of androgen deficiency and infertility.</p><p><b>METHODS</b>We retrospectively analyzed 12 cases of testis homotransplantation.</p><p><b>RESULTS</b>Surgical success was achieved in 11 cases, all with a significantly increased level of serum testosterone, and markedly improved secondary sex characteristics and sexual function.</p><p><b>CONCLUSION</b>Testis homotransplantation is highly effective for the treatment of androgen deficiency in males, but has little effect on spermatogenesis.</p>

Construction and expression of eukaryotic recombinant of mouse fertilin beta subunit in HEK293 cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To construct the eukaryotic expression recombinant pSG. SS. C3d3. YL-Fbeta and analyze the expression of mouse fertilin beta subunit in HEK293 cells.</p><p><b>METHODS</b>The cDNA fragment expressing the disintegrin domain of mouse fertilin beta was obtained by PCR, and then inserted into the eukaryotic plasmid pSG. SS. C3d3. YL to get recombinant plasmid pSG. SS. C3d3. YL-Fbeta, which was transfected into the HEK293 cell line to express the target protein Fbeta after identified by restriction enzyme digestion. And then Fbeta was detected by indirect immunofluorescence through confocal laser scanning microscopy, Western blot, immunohistochemistry staining and flow cytometry assay.</p><p><b>RESULTS</b>The recombinant vector pSG.SS.C3d3. YL-Fbeta could express Fbeta in HEK293 cells.</p><p><b>CONCLUSION</b>The expression of Fbeta in eukaryotic cells provides a foundation for further researches on the effect of high F, expression on fertilization process.</p>

Long-term survival of human spermatogonial stem cells in vitro and its functional identification / 中华男科学杂志

<p><b>OBJECTIVE</b>The culture of human spermatogonial stem cells (SSC) has not been studied in detail yet. Here we tried to explore the optimized culture method of human SSC by using several different co-culture systems.</p><p><b>METHODS</b>The alpha6 +Thy-1 +c-kit- cells acquired by the immunomagnetic beads sorting technique were cultured in different co-culture systems. Their morphological, biological characteristics and survival rates were intensively observed by microscopic or immunocytochemical assay. The long-term survival rate of human SSC during culture period was evaluated by germ cell transplantation technique.</p><p><b>RESULTS</b>The alpha6 +Thy-1 +c-kit- cells could stably survive in the DMEM and DMEM/F12 mediums with fetal bovine serum (FBS) on feeder layer. The survival rates within 1 week were more than 90%. The long-time culture showed the cells were gradually attached on the surface of Sertoli cells by the manner of scattered single cell or accumulated masses. Part of the SSC became more tightly attachment with Sertoli cells or mounted among the Sertoli cells. They could survive or even proliferate for more than 3 months in vitro. Germ cells transplantation study showed that some alpha6 +Thy-1 +c-kit- cells labeled by PKH26 could resided on the basal membrane of seminiferous tubule of nude mice, appearing as single or coupled cells 2 months later after transplantation. The function evaluation of the cultured cells by counting the fluorescent cells in the seminiferous tubule showed 54.9% and 9.2% of SSC in the alpha6 +Thy-1 +c-kit- cells were still remained after cultured for 2 and 4 weeks, respectively.</p><p><b>CONCLUSION</b>Human SSC could maintain survival in vitro for more than 3 months, but it was still need to seek for a more optimized and successful culture system for its efficient expansion and proliferation. Thus it will open up a wide prospect for the understanding of the biology of human SSC and the treatment of male sterility.</p>

Preliminary screening of specific surface markers of human spermatogonial stem cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the specific surface markers for the isolation and purification of human spermatogonial stem cells (SSC).</p><p><b>METHODS</b>Specific markers of human SSC were screened and identified in fetal and adult testes by immunohistochemical assay, using HSC markers c-kit, Thy-1 and human ES integrins.</p><p><b>RESULTS</b>In human adult testes, the alpha6 integrin extensively and significantly expressed on the surface of most of the germ cells in the seminiferous tubule, and beta1 integrin mainly expressed on the surface of the germ cells residing on or near the basal membrane in the seminiferous tubule. Thy-1 scattering expressed on the surface of some cells of the basal membrane, and on some Leydig cells as well. The three antigen markers expressed on the SSC of human adult testes specifically to some extent. SSEA-1 specifically expressed on the surface of the gonocytes in the fetal testes.</p><p><b>CONCLUSION</b>The alpha6 and beta1 integrins and Thy-1 may be used for the SSC isolation as positive markers. SSEA-1 can be used as an identification marker for the fetus SSC.</p>

Proliferative response of human prostate cancer cell to hormone inhibited by androgen receptor antisense RNA / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The failure of endocrine treatment for advanced prostate cancer might be related to aberrant activation of androgen receptor (AR). Prostate cancer cell line LNCaP contains AR that can be activated by androgen, estrogen and progesterone. This study was set to investigate the effects of antisense AR RNA on growth of LNCaP cultured in medium containing varied concentrations of R1881, 17beta-estradiol, and progesterone, respectively.</p><p><b>METHODS</b>LNCaP cells transfected with antisense AR RNA retroviral vector pL-AR-SN were designated as LNCaPas-AR. LNCaP cells containing empty vector pLXSN served as LNCaPNeo. LNCaP and LNCaPNeo were taken as controls. In vitro cell growth assay, proliferative cells of LNCaP and tranfected LNCaPs were counted by typan staining when they cultured with synthetic androgen R1881, 17beta-estradiol, and progesterone, respectively.</p><p><b>RESULTS</b>Growth of LNCaPas-AR was inhibited significantly (P < 0.05) compared with that of LNCaP and LNCaPNeo at 1 nmol/L R1881, 10 nmol/L 17beta-estradiol, and 1 nmol/L progesterone, respectively. No difference was seen between LNCaP and LNCaPNeo (P > 0.05). Microscopic observation showed that LNCaP and LNCaPNeo cells grew well, but only few LNCaPas-AR cells were alive.</p><p><b>CONCLUSIONS</b>Our observations indicate that antisense AR RNA retroviral vector pL-AR-SN could change androgen-independent characteristics of LNCaP cells, which might shed some novel insights into the treatment of androgen-independent prostate cancer.</p>