Exploration on Determination and Quantity Transfer of Index Components in Benchmark Samples of Erdongtang / 中国实验方剂学杂志

ObjectiveTo establish the determination for index components in benchmark samples of Erdongtang， and clarify the content and transfer rate rages of index components in 15 batches of benchmark samples， and to explore the quantity transfer of index components of decoction pieces to benchmark samples. MethodFifteen batches of benchmark samples were prepared， the contents of mangiferin， baicalin and glycyrrhizic acid were determined by high performance liquid chromatography （HPLC）-diode array detector （DAD）， the mobile phase was acetonitrile （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-10 min， 10%-17%A； 10-25 min， 17%-19%A； 25-28 min， 19%-25%A； 28-45 min， 25%-33%A； 45-46 min， 33%-45%A； 46-60 min， 45%-55%A）， detection wavelength was set at 254 nm. Contents of timosaponin BⅡ and the sum of protoneodioscin and protodioscin were determined by HPLC-evaporative light scattering detector （ELSD）， the mobile phase was acetonitrile （A）-water （B） for gradient elution （0-20 min， 24%A； 20-25 min， 24%-27%A； 25-33 min， 27%-28%A； 33-36 min， 28%-90%A； 36-41 min， 90%-24%A）. ResultThe methodological verification of the established method was good， which could be used for determination of five index components in benchmark samples. The content ranges of mangiferin， baicalin， glycyrrhizic acid， timosaponin BⅡ， and the sum of protoneodioscin and protodioscin in 15 batches of benchmark samples of Erdongtang were 0.14%-0.23%， 2.40%-3.37%， 0.07%-0.44%， 0.43%-0.95%， and 0.15%-0.47%， the transfer rate ranges of them were 33.90%-52.15%， 84.46%-105.61%， 22.59%-93.86%， 38.07%-61.43%， and 53.28%-96.11%， respectively. ConclusionThe consistencies of transfer rate of mangiferin， baicalin， timosaponin BⅡ and the sum of protoneodioscin and protodioscin （except glycyrrhizic acid） between decoction pieces and benchmark samples of Erdongtang are good， indicates that the transfer rates of 4 index components are stable during the preparation process of benchmark samples， which can provide data support for research and development of the compound preparation of this formula.

The mechanism of androgen independent signaling pathway in castration-resistant prostate cancer and the research progress on related drugs / 药学学报

Prostate cancer is one of the common malignant tumors of male urogenital system, and the incidence of prostate cancer in China has increased significantly in the past decade. At present, endocrine therapy based on androgen blockade is the main method of clinical treatment except radical surgery and radiotherapy/chemotherapy for prostate cancer. However, the clinical benefit can only be obtained in the early stage of treatment, and nearly 90% of patients will develop to the castration resistance, and among them, nearly 90% of patients will have bone metastasis. The quality of life decreases sharply with the progression of disease for patients. In addition to the androgen signal pathway, studies have shown that many other oncogenic signal pathways have involved in the development of castration resistance, including classic cancer signaling pathways, immune and inflammatory signaling pathways, etc. Understanding the mechanism of androgen independent signal pathway in the formation of castration resistance will help to understand the off-target effect of androgen blocking therapy and introduce new treatment targets or strategies to get rid of the "no drug available" dilemma for clinical treatment of castration resistance.

Association of Overlapped and Un-overlapped Comorbidities with COVID-19 Severity and Treatment Outcomes: A Retrospective Cohort Study from Nine Provinces in China / 生物医学与环境科学（英文）

Objective@#Several COVID-19 patients have overlapping comorbidities. The independent role of each component contributing to the risk of COVID-19 is unknown, and how some non-cardiometabolic comorbidities affect the risk of COVID-19 remains unclear.@*Methods@#A retrospective follow-up design was adopted. A total of 1,160 laboratory-confirmed patients were enrolled from nine provinces in China. Data on comorbidities were obtained from the patients' medical records. Multivariable logistic regression models were used to estimate the odds ratio ( @*Results@#Overall, 158 (13.6%) patients were diagnosed with severe illness and 32 (2.7%) had unfavorable outcomes. Hypertension (2.87, 1.30-6.32), type 2 diabetes (T2DM) (3.57, 2.32-5.49), cardiovascular disease (CVD) (3.78, 1.81-7.89), fatty liver disease (7.53, 1.96-28.96), hyperlipidemia (2.15, 1.26-3.67), other lung diseases (6.00, 3.01-11.96), and electrolyte imbalance (10.40, 3.00-26.10) were independently linked to increased odds of being severely ill. T2DM (6.07, 2.89-12.75), CVD (8.47, 6.03-11.89), and electrolyte imbalance (19.44, 11.47-32.96) were also strong predictors of unfavorable outcomes. Women with comorbidities were more likely to have severe disease on admission (5.46, 3.25-9.19), while men with comorbidities were more likely to have unfavorable treatment outcomes (6.58, 1.46-29.64) within two weeks.@*Conclusion@#Besides hypertension, diabetes, and CVD, fatty liver disease, hyperlipidemia, other lung diseases, and electrolyte imbalance were independent risk factors for COVID-19 severity and poor treatment outcome. Women with comorbidities were more likely to have severe disease, while men with comorbidities were more likely to have unfavorable treatment outcomes.

Classification of uterine fibroids and strategy of clinical treatment / 中国实用妇科与产科杂志

Uterine fibroid is a common gynecological disease,patients with which would present no symptoms or severe symptoms based on the location and size of focus.According to their relationship with uterine cavity and serosa,uterine fibroids can be classified into several types.In clinical practice,different measures should be taken depending on the type and size of fibroids as well as the age,fertility desire,reproductive function and symptoms of patients.This paper elaborates the classification of uterine fibroids and corresponding strategy of clinical treatments.

Reproductive outcomes of fertility-sparing treatment for gynecologic malignancies / 中国实用妇科与产科杂志

Cervical cancer,endometrial cancer and ovarian cancer are common gynecologic malignancies.Recently,with the younger onset age and delay of childbearing,the fertility-sparing treatment has been increasingly used.In the fertility-sparing treatment for gynecologic malignancies,we should protect the fertility of patients without increasing tumor recurrence,which requires the multidisciplinary cooperation to develop individualized treatment strategy,provide optimal reproductive guidance and finally improve the reproductive outcomes.

Network analysis of microRNAs, transcription factors, and target genes involved in axon regeneration / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Axon regeneration is crucial for recovery from neurological diseases. Numerous studies have identified several genes, microRNAs (miRNAs), and transcription factors (TFs) that influence axon regeneration. However, the regulatory networks involved have not been fully elucidated. In the present study, we analyzed a regulatory network of 51 miRNAs, 27 TFs, and 59 target genes, which is involved in axon regeneration. We identified 359 pairs of feed-forward loops (FFLs), seven important genes (Nap1l1, Arhgef12, Sema6d, Akt3, Trim2, Rab11fip2, and Rps6ka3), six important miRNAs (hsa-miR-204-5p, hsa-miR-124-3p, hsa-miR-26a-5p, hsa-miR-16-5p, hsa-miR-17-5p, and hsa-miR-15b-5p), and eight important TFs (Smada2, Fli1, Wt1, Sp6, Sp3, Smad4, Smad5, and Creb1), which appear to play an important role in axon regeneration. Functional enrichment analysis revealed that axon-associated genes are involved mainly in the regulation of cellular component organization, axonogenesis, and cell morphogenesis during neuronal differentiation. However, these findings need to be validated by further studies.

Effect of laser artificial shrinkage on pregnancy outcome in vitrification of human expanded blastocysts / 西安交通大学学报(医学版)

Objective To investigate the effect of laser artificial shrinkage(LAS)on pregnancy outcome in vitrification of human expanded blastocysts.Methods We selected 3859 frozen-thawed blastocyst-stage embryo transfers from January 2014 to December 2015.The transfers were divided into LAS group(n=3 176)and non-LAS group(n=683),which were then subdivided into ＜36 y subgroup and ≥36 y subgroup according to their age.Main outcomes measures were thawing rate,implantation rate and clinical pregnancy rate.Results Thawing rate, clinical pregnancy rate and implantation rate were 97.32%(5 453/5 603),66.81%(2 118/3 170),and 53.55%(2 912/5 438)in LAS group.In non-shrink group,they were 95.13%(1 173/1 233),62.70%(427/681),and 49.74%(582/1 170),which did not significantly differ from those in the former group(P＜0.05).Further analysis of the subgroups showed that thawing rate was significantly higher in LAS group than in non-shrink group of patients＜36 y(97.27% vs.95.33%;P＜0.05).Thawing rate and biochemical pregnancy rate were significantly higher in LAS group than in non-shrink group in patients ≥36 y(97.75% vs.93.66%;65.45% vs.50.65%,P＜0.05). Cancellation rate was not significantly different between the two groups(0.19% vs.0.29%, P ＞ 0.05). Conclusion LAS technique can increase thawing rate,clinical pregnancy rate and implantation rate before cryopreservation of blastocysts.

The rate of morphologically normal sperm does not affect the clinical outcomes of conventional IVF in patients with one retrieved oocyte / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the influence of the rate of morphologically normal sperm (MNS) on the clinical outcomes of conventional in vitro fertilization (IVF) in patients with one retrieved oocyte.</p><p><b>METHODS</b>From January 2013 to January 2015, a total of 256 couples with one retrieved oocyte underwent conventional IVF in our center. According to the rate of MNS, the patients were divided into two groups: MNS < 4% (134 cycles) and MNS ≥ 4% (122 cycles). We compared the rates of no transferrable embryo cycles, fertilization, cleavage, normal fertilization, abnormal fertilization, high-quality embryo and transferrable embryo between the two groups. A total of 75 fresh embryo transfer cycles were performed, 43 in the MNS < 4% group and the other 32 in the MNS ≥ 4% group. We also compared the rates of implantation, clinical pregnancy and abortion between the two groups.</p><p><b>RESULTS</b>There were no statistically significant differences between the two groups in the rates of no transferrable embryo cycles, fertilization, cleavage, normal fertilization, abnormal fertilization, high-quality embryo and transferrable embryo (P > 0.05). The rates of implantation, clinical pregnancy and abortion exhibited no remarkable differences either in the fresh embryo transfer cycles between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>The rate of MNS does not affect the clinical outcomes of conventional IVF in patients with one retrieved oocyte.</p>

Key nodes of a microRNA network associated with the integrated mesenchymal subtype of high-grade serous ovarian cancer / 癌症

Metastasis is the main cause of cancer mortality. One of the initiating events of cancer metastasis of epithelial tumors is epithelial-to-mesenchymal transition (EMT), during which cells dedifferentiate from a relatively rigid cell structure/morphology to a flexible and changeable structure/morphology often associated with mesenchymal cells. The presence of EMT in human epithelial tumors is reflected by the increased expression of genes and levels of proteins that are preferentially present in mesenchymal cells. The combined presence of these genes forms the basis of mesenchymal gene signatures, which are the foundation for classifying a mesenchymal subtype of tumors. Indeed, tumor classification schemes that use clustering analysis of large genomic characterizations, like The Cancer Genome Atlas (TCGA), have defined mesenchymal subtype in a number of cancer types, such as high-grade serous ovarian cancer and glioblastoma. However, recent analyses have shown that gene expression-based classifications of mesenchymal subtypes often do not associate with poor survival. This "paradox" can be ameliorated using integrated analysis that combines multiple data types. We recently found that integrating mRNA and microRNA (miRNA) data revealed an integrated mesenchymal subtype that is consistently associated with poor survival in multiple cohorts of patients with serous ovarian cancer. This network consists of 8 major miRNAs and 214 mRNAs. Among the 8 miRNAs, 4 are known to be regulators of EMT. This review provides a summary of these 8 miRNAs, which were associated with the integrated mesenchymal subtype of serous ovarian cancer.

Expression of peptidylarginine deiminase 4 and protein tyrosine phosphatase nonreceptor type 22 in the synovium of collagen-induced arthritis rats / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To study the expression level of peptidylarginine deiminase 4 (PADI4) and protein tyrosine phosphatase nonreceptor type 22 (PTPN22) in the synovium of rat model of collagen-induced arthritis, and to explore their possible therapeutic role in rheumatoid arthritis.</p><p><b>METHODS</b>Thirty-two female Wistar rats weighing 100±20 g were randomly assigned into 3-week collagen-induced arthritis (CIA) model group (n=8), 4-week CIA model group (n=8), 6-week CIA model group (n=8), and the control group (n=8). The body weight changes of each group were recorded. The expression levels of PADI4 and PTPN22 were detected and compared by the methods of immunohistochemical staining and Western blot.</p><p><b>RESULTS</b>Arthritis of rat began to form 14 days after sensitization and the joint swelling reached peak at 28 days. The weights of the rats slowly grew both in CIA model groups and the control group. Immunohistochemical staining results showed that the positive expression of PADI4 and PTPN22 was mainly located in cartilage peripheral mononuclear cells, the cytoplasm of infiltrated cells, and bone marrow cavity. There were significant differences in the optical density of PADI4 and PTPN22 among CIA model groups and the control group (PADI4, 0.2898±0.012, 0.2982±0.022, 0.2974±0.031, 0.2530±0.013 in 3-week CIA model, 4-week CIA model, 6-week CIA model and control groups; PTPN22, 0.2723±0.004, 0.2781±0.010, 0.2767±0.008, 0.2422±0.019; all P <0.05). The expression bands of PADI4 were observed in Western blot 3 weeks after initial immunization, the thickest in the 4th week, and decreased in the 6th week. The expression bands of PTPN2 were observed at all the time points, with no obvious time-dependent trend.</p><p><b>CONCLUSIONS</b>PADI4 and PTPN22 are obviously correlated with CIA in rat model. PADI4 is expressed at early stage of the disease, while the expression of PTPN22 sustains throughout the course.</p>

25-Hydroxyvitamin D is closely related with the function of the pancreatic islet beta cells

This study is to investigate the relationship between 25-Hydroxyvitamin D [25-OH-D] and pancreatic islet beta cell function under different glucose tolerance statuses in China. Totally, 180 patients with type 2 diabetes mellitus [DM group], 178 patients with impaired fasting glucose/impaired glucose tolerance [IFG/IGT group], and 160 normal control subjects [NGT group] were included to measure their body parameters and biochemical parameters. In oral glucose tolerance test, fasting serum 25-OH-D was assessed by the enzyme-linked immunosorbent assay [ELISA]. Homeostasis model assessment for insulin resistance [Homa-IR], insulin acuity index [IAI], beta -cell function index [Homa-BCF] as well as secretion index [IS] were determined. The levels of 25-OH-D, IAI and Homa-BCF in the DM group and IFG/IGT group were significantly lower than that in NGT group [P < 0.05]. Homa-IR in DM group and IFG/IGT group was significantly higher than that in the NGT group [P < 0.05]. Pearson correlation analysis and partial correlation analysis showed that 25-OH-D was positively correlated with fasting insulin [FINS] and Homa-BCF [P < 0.05]. Multiple stepwise regression analysis showed that 25-OH-D was one of the influential factors of pancreatic islet beta cell function in patients with type 2 diabetes mellitus. Our results suggest that 25-OH-D is closely related with the function of the pancreatic islet beta cells and is one of the influential factors of pancreatic islet beta cell function

Modified super-long down-regulation protocol improves fertilization and pregnancy in patients with poor ovarian responses / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The successful end-point of in vitro fertilization (IVF) treatment is for a woman to give live birth. This outcome is based on various factors including adequate number of retrieved eggs. Failure to recruit adequate follicles, from which the eggs are retrieved, is called a "poor response". How to improve the clinical pregnancy rates of poor responders was one of the tough problems for IVF.</p><p><b>METHODS</b>The study involved 51 patients who responded poorly to high dose gonadotropin treatment in their previous cycles at our reproductive center, between April 2010 and February 2012. The previous cycle (group A) received routine long protocol; the subsequent cycle (group B) received modified super-long down-regulation protocol. The primary outcome of the study was the number of oocytes fertilized. The increase in the pregnancy rate was the secondary outcome. Differences between the groups were assessed by using Student's t test and c(2) test where appropriate.</p><p><b>RESULTS</b>The patients' average age was (36.64 ± 3.85) years. The mean duration of ovarian stimulation cycles of the group A patients was longer than those of the group B patients. The total dose of follicle-stimulating hormone (FSH) was significantly lower in the subsequent cycle. The peak value of serum estradiol on human chorionic gonadotrophin (hCG) day was lower in group A as compared with group B. The number of metaphase II oocytes recovered was significantly higher in group B. The cleavage rate in group A was significantly lower than in group B, 49 patients in group B reached embryo transfer stage, while 46 patients in group A reached this stage. Patients in group B received significantly more embryos per transfer as compared with group A. More pregnancies and more clinical pregnancies with fetal heart activity were achieved in group B.</p><p><b>CONCLUSIONS</b>This comparative trial shows that poor responder women undergoing repeated assisted reproduction treatment using modified super-long down-regulation protocol achieve more oocytes, leading to higher fertilization rate, compared to women receiving routine long protocol. Our study also showed that clinical pregnancy rate was significantly improved.</p>

Analysis of human papillomavirus type 61, 83 and 84 MY fragments and preparation of standard samples / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the mutations in nucleotide and amino acid level in HPV61, 83 and 84 Shanxi isolates.</p><p><b>METHODS</b>Amplified fragments of HPV61, 83 and 84 from human papillomavirus (Human papillomavirus, HPV) molecular epidemiologic survey of Shanxi Province using HPV consensus primers MY09/11 were cloned in pMD18-T vector, and the plasmids were sequenced, then nucleotide sequences and amino acid sequences were analyzed.</p><p><b>RESULTS</b>HPV61 and HPV83 isolates were consistent with reference strains U31793 and AF151983 in nucleotide sequences; four mutations of nucleotide (C6760T, T6931C, T6951C and C6987A) were found in HPV84 isolate compared with reference strain AF293960, among which C6987A resulted in D441E and the amplified sensitivity of standard sample of HPV61 using primers MY09/11 was higher than that of HPV83 and 84.</p><p><b>CONCLUSION</b>HPV61 and HPV83 isolates were consistent with reference strains, four mutations of nucleotide and one mutation of amino acid were found in HPV84,the amplified sensitivity of standard sample of HPV61 using primers MY09/11 was the highest among those three isolates.</p>

Construction and expression of growth differentiation factor-5 gene prokaryotic expression plasmid and its cartilage induction activity in vivo / 中国组织工程研究

BACKGROUND: Growth differentiation factor-5 (GDF-5) plays an important role in the development and formation of cartilage, extremities, and joints, which is a widely used joint development marker.OBJECTIVE: To express mature peptide of human GDF-5 in E. coil by the way of genetic engineering, and to explore the inductive activity of recombinant protein in vivo.DESIGN, TIME AND SETTING: The observation experiment based on gene was performed at the Analysis and Testing Center of Southern Medical University from January to June 2006.MATERIALS: Human fetus cartilage tissue was harvested from Department of Gynaecology and Obstetrics, and the consent was obtained from the family. Ten KM mice were purchased from experimental animal center of Southern Medical University, half male and half female, weighing 18-22 g, aged 6-8 weeks.METHODS: The hGDF-5 gene encoding mature peptide was gained by RT-PCR from the total RNA which was extracted from fetus cartilage tissues, and was inserted into the pET22b(+) vector to construct recombinant prokaryotic expression plasmid pET22b(+)-GDF5, which was transformed into E. coil BL-21 to be expressed after IPTG induction. Proteins of interest were purified with sepharose chelated with nickel ions (Ni2+) and then implanted in mouse hindlimb muscle to evaluate the biological activities by routine hematoxylin-eosin staining.MAIN OUTCOME MEASURES: The expression, sequencing of target gene was observed by agarose gel electrophoresis, and the protein expression was detected by SDS-PAGE electrophoresis, meanwhile, the GDF5-inducing activity was evaluate by histological observation.RESULTS: RT-PCR product was about 350 bp in length, which was confirmed by double enzyme digestion of the recombinant plasmid, sequencing result was in agreement with the reported hGDF-5 sequence in Genbank. SDS-PAGE analysis showed a conspicuous band representing a new foreign protein with relative molecular mass of approximately 14 KD after induced expressioin. Cartilage tissues were formed in the mouse muscle where the purified proteins were implanted. CONCLUSION: The integral human GDF-5 mature peptide gene was cloned successfully from human fetus cartilage tissue and a high-yield expression was achieved in E. coli, the pudfied protein has chondrogenic activities in vivo.

Teratozoospermia affects in vitro fertilization outcome / 中华男科学杂志

<p><b>OBJECTIVE</b>To analyze the possible causes of total fertilization failure (TFF) in conventional in vitro fertilization (IVF).</p><p><b>METHODS</b>We included in this study 552 couples who accomplished the first conventional IVF cycle in our center from January 2007 to August 2008. All the males met the Kruger/Tygerberg criteria, with the teratozoospermia rate < 96% in the previous 12 months and the postwash motile sperm > 1 million on the day of egg retrieval. The eggs were fertilized totally by conventional IVF.</p><p><b>RESULTS</b>Of the total number, 515 couples got zygotes by conventional IVF, with a fertilization rate of 78.67%, and 37 suffered TFF. The rates of primary infertility and teratozoospermia were significantly higher in the TFF group than in the fertilized group (P < 0.05). There were no statistically significant differences between the two groups in the number of controlled ovarian hyperstimulation (COH) days, the number of oocytes retrieved, the dose of gonadotropin (Gn) used, the age of the couples, the length of protocols and the rate of oligoasthenozoospermia.</p><p><b>CONCLUSION</b>Intracytoplasmic sperm injection (ICSI) can be considered for at least some of the oocytes to avoid unnecessary fertilization failure in teratozoospermia patients by Kruger/Tygerberg strict criteria.</p>

Effects of low arsenic sub-chronic exposure on blood general examination index in rabbit / 中国地方病学杂志

Objective To observe the effect of low arsenic sub-chronic exposure on blood routine test index in rabbits to pave a way for screening early injury of the low arsenic exposure. Methods Twelve healthy male rabbits were randomly divided into four groups. They were administrated with As at the concentration of 0(control), 10, 50 and 250 μg/L in the drinking water. Blood samples were collected from the vein of the ear edge in 8 weeks, and blood test routine including leukocyte (WBC), lymphocyte (LYM), lymphocyte percentage (LYM%), neutrophil (GRA), neutrophil percentage (GRA%), monocyte (MON), monocyte percentage (MON%), red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin content (MCH), mean corpuscular hemoglobin concentration(MCHC), RDW, platelets(PLT), mean platelet volume(MPV), platelet hematocrit(PCT) and platelet distribution width (PDW), were detected by the ABX-60 hemocyte analyzer. Results Compared with the control group, the WBC, GRA and GRA% increased in 0, 50 and 250 μg/L groups, but there was no significance(P > 0.05). PLT and MPV had a statistical significance in 4 groups(F = 4.07,4.20, all P < 0.05). Compared with the control group[(292.00±16.97)×109/L, (7.10±0.99)fL], PLT decreased in the 250 μg/L group [(221.33±22.50)×109/L] and MPV decreased in the 50μg/L group [(5.57±0.46)fL] significantly (P < 0.05). The other index didn't change obviously. Conclusions Sub-chronic low level arsenic exposure may lead to the change in the blood system. The blood routine test may be considered for early injury of the arsenic poisoning.

Effect of local viral transfer of interleukin 10 gene on a rabbit arthritis model induced by interleukin 1beta / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Interleukin 1beta (IL-1beta) is the principal mediator in the pathogenesis of rheumatoid arthritis. Continuous injection of interleukin 1beta (IL-1beta) into the knee articular cavities of animals can induce models that resemble rheumatoid arthritis. The objective of this study was to evaluate the feasibility of local recombinant retrovirus viral interleukin 10 (rRV-vIL-10) gene transfer treatment of a rabbit model of arthritis induced by IL-1beta.</p><p><b>METHODS</b>An hIL-1beta-induced rabbit rheumatoid arthritis model was established using the MFG-hIL-1beta-neo-HIG-82 cell line, which is capable of continuous secretion of hIL-1beta. After transfecting the rabbit synovial fibroblast cell line (MFG-hIL-1beta-neo-HIG-82) with rRV-vIL-10, G418 was then added to identify the positive clone. The rRV-vIL-10 positive clone was injected into the established rabbit rheumatoid arthritis model through intra-articular injection. Successful gene transfer was determined by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. The levels of IL-1beta before and after treatment were determined by enzyme- linked immunosorbent assay.</p><p><b>RESULTS</b>Retrovirus vector was an effective vector both to synoviocytes in vitro and synovium tissue in vivo as confirmed by RT-PCR and immunohistochemistry. The rabbit arthritis model treated with rRV-vIL-10 showed a dramatic remission of arthritis and a decline in the level of cytokines such as IL-1beta.</p><p><b>CONCLUSIONS</b>Retrovirus-mediated transfection of vIL-10 successfully transferred the gene into rabbit synovium ex vivo and was able to suppress intra-articular inflammation response to IL-1beta.</p>

Effects of experimental left varicocele on SPAG11 mRNA and SPAG11E in the testis and epididymis of adolescent rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of experimental left varicocele (ELV) on the expression of sperm associated antigen 11 (SPAG11) mRNA and its protein isomer SPAG11E in the testis and epididymis of adolescent rats, and to explore the mechanism of infertility caused by varicocele.</p><p><b>METHODS</b>The experimental left varicocele model was established in the adolescent male Sprague-Dawley rats. Two and 4 weeks after the operation, the changes of SPAG11 mRNA and SPAG11E expression in the testis and epididymis were detected using immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The expected product of SPAG11 367 bp amplified by RT-PCR was detected only in the epididymis. SPAG11E protein was observed mainly in the acrosomal vesicles and acrosome of round and elongating spermatids of the seminiferous epithelium, in the cytoplasm of Leydig cells, and in the supranuclear region of principle cells and stereocilia of the epididymal epithelium. Imaging and statistical analysis showed that SPAG11 mRNA and SPAG11E protein expressions in the left epididymis of the 2- and 4-week ELV groups presented a remarkable decrease (P < 0.05 or P < 0.01) compared with the right side and the corresponding control group, and the same decreased change in the left epididymis (P < 0.05 or P < 0.01) and an obvious reduction of SPAG11E immunopositive reaction in the right epididymis (P < 0.01) were noted in the 4-week group as compared with the 2-week group. No statistical difference of SPAG11E expression in the bilateral testes was found (P > 0.05) between the ELV group and the control, as well as between the 2- and 4-week ELV groups.</p><p><b>CONCLUSION</b>SPAG11 is a specific gene expressed in the epididymis. The localization and expression of SPAG11E exhibited a region- and cell-specific pattern in both the testis and epididymis of adolescent rats. The expression levels of both SPAG11 mRNA and SPAG11E protein altered obviously in ELV rats. The results suggest that SPAG11 may not only play an important role in spermatogenesis and sperm maturation, but also be associated with varicocele-induced male infertility or subfertility.</p>

Effect of RNA interference for Lon gene silencing on growth and apoptosis of human breast cancer MCF7 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the role of Lon gene in tumor cell proliferation, apoptosis and cell stress response.</p><p><b>METHODS</b>Small interfering RNAs (smRNAs) for Lon gene were designed using Ambion software and synthesized. The recombinant plasmid pSilencer U6 2.1/Lon was constructed with the smRNAs and pSilencer U6 2.1, followed by transfection into MCF7 cells via Lipofectamine(TM) 2000. The positive cLones were detected by RT-PCR 24 h after cell transfection. The transfected MCF7 cells were then subjected to cisplatin treatment, ultraviolet (UV) exposure and heat stress, respectively, after which the cells growth was tested with MTT assay and the measurements were plotted against time or concentration depending on the treatment administered. Apoptosis of MCF7 cells following the treatments was measured with flow cytometry.</p><p><b>RESULTS</b>The mRNA of Lon gene was downregulated in cells transfected with the recombinant plasmid pSilencer U6 2.1-Lon, and RT-PCR fail to detect the specific band of Lon as could be detected in untransfected and mock-transfected MCF7 cells. MTT assay showed that pSilencer U6 2.1-Lon transfection resulted in reduced cell proliferation capacity. Stress response test revealed that MCF7 cells with Lon gene down-regulation enhanced cell sensitivity for UV and cisplatin, which was not observed for non-transfected or mock transfection group. The same changes were also observed for heat stress exposure at 41 degrees Celsius;, but not at 43 degrees Celsius; or 45 degrees Celsius;. Increased cell apoptosis rate from (1.14-/+0.79)% to (22.47-/+3.15)% occurred following pSilencer U6 2.1-Lon transfection of the cells.</p><p><b>CONCLUSIONS</b>Lon gene can be significantly downregulated by introduction of siRNA in MCF7 cells to result in enhanced sensitivity of MCF7 cells to UV, cisplatin and heat stress.</p>

Expression of human epididymis protein 2beta1 in the testis and epididymis of adolescent male rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the expression of human epididymal secretary protein 2 isoform human epididymal protein 2beta1(HE2beta1) in the testis and epididymis of adolescent male rats along with its significance.</p><p><b>METHODS</b>Immunohistochemical staining was used to detect the expression and localization of HE2beta1 in the testis and epididymis of 15 adolescent SD rats.</p><p><b>RESULTS</b>HE2beta1 immunoreactive staining was detected in the testis and epididymis. In the epithelia of the epididymal duct, HE2beta1 expressed mainly in the supranuclear region of the principle cells and the basement membrane of some epithelial cells; there were no immunostaining in the n clear cells, halo cells and basal cells. The immunopositive reaction was detected, weak in the distal caput, strong in the proximal, middle corpus and the cauda, but negative in the initial segment. Immunopositive results of HE2beta1 were also observed in some of the nuclei of spermatogonia and Sertoli cells with negatively-stained cytoplasm.</p><p><b>CONCLUSION</b>Immunohistochemical staining is a fairly sensitive method for detecting HE2beta1 expression. The localization and expression level of HE2beta1 in the genital duct of adolescent male rats exhibited a region- and cell-specific expression pattern, which suggests that HE2beta1 may play an important role in spermatogenesis, maturation and epididymal epithelial innate defense mechanisms.</p>