OBJECTIVETo investigate the feasibility and clinical application value of selecting viable spermatozoa by noncontact diode laser.

METHODSWe obtained immotile spermatozoa from 2 infertile men with obstructive azoospermia or severe asthenospermia and selected viable spermatozoa using a single laser shot at the sperm tail. Those that responded to the laser shot by a curling reaction of the tail were regarded as presumably viable and used for intracytoplasmic sperm injection (ICSI).

RESULTSThe mean fertilization rate was 88.89% after ICSI with the laser-selected viable spermatozoa. Both of the embryo transfers resulted in a single pregnancy.

CONCLUSIONNoncontact diode laser is a useful alternative for the assessment of sperm viability, which may help to achieve successful pregnancy.

Embryo Transfer ; Female ; Fertilization ; Humans ; Infertility, Male ; therapy ; Male ; Pregnancy ; Pregnancy Outcome ; Sperm Injections, Intracytoplasmic ; Sperm Motility ; Sperm Tail ; physiology

OBJECTIVE Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited disease with a high morbidity around 1/1000-1/400, characterized by progressive enlargement of fluid-filled cysts derived from renal tubular epithelial cells. Massive cysts gradually compress renal parenchyma destroying normal renal structures and compromising renal functions. Unfortunately, it will cause end-stage renal disease in most of the patients but without effective therapy now, who have to live on hemodialysis or kidney transplantation. Based on this present situation, it is of great significance to find early intervention to inhibit renal cyst development. The projective of this study was to investigate whether Ganoderma triterpenes (GT) can inhibit renal cyst development and study the related mechanism. METHODS and RESULTS First, we used MDCK cyst model, cultivated MDCK cells in vitro to form fluid-filled cysts surrounded by monolayer cells. GT inhibited MDCK cyst formation significantly, and inhibited cyst enlargement dose-dependently proving GT cyst inhibition in vitro. Then we used an embryonic kidney cyst model, wile-type mice kidneys were taken out on embryonic day 13.5 to form renal cysts stimulated with 8-Br-cAMP. GT inhibited embryonic kidney cyst development significantly in a dose-dependent and reversible manner proving GT cyst inhibition at organ level. Furthermore, we used two ADPKD mouse models with severe cystic kidney disease phenotypes. GT dramatically inhibited renal cyst development, decreased ADPKD mouse kidney volume and the cyst index inside proving GT cyst inhibition in vivo. By Western blot, we proved GT down-regulated Ras/MAPK signal pathway without detectable effect on mTOR signal pathway both in MDCK cells and two ADPKD mouse kidneys. CONCLUSION GT retard renal cyst development both in vitro and in vivo significantly. The related mechanisms were involved in GT promoting renal tubular epithelial cell differentiation, down-regulating intracellular cAMP level and Ras/MAPK signal pathway.

This study aims to analyze the molecular epidemiological characteristics of HIV-1 strains prevailing among men who have sex with men (MSM) in Beijing, China. The pol gene fragments from 250 newly diagnosed HIV-1-infected MSM individuals during 2006-2010 in Beijing were amplified by RT-nested PCR, sequenced, and phylogenetically analyzed. HIV-1 pol gene from 189 individuals were amplified and analyzed; 81 (42. 9%), 3 (1. 6%), 2 (1.0%), 88 (46. 6%), and 15 (7.9%) individuals were infected with HIV-1 subtypes B, B', C, CRF01_AE, and CRF07_BC, respectively. The subtypes B and CRF01_AE could both be grouped into two clusters, and CRFO7_BC strains shared high homology and were presumed to originate from a common ancestor. The HIV-1 circulating in MSM in Beijing had a lower genetic diversity than in heterosexuals. The HIV-1 epidemic (2006-2010) in MSM in Beijing was actually a rapid spread of HIV-1 CRF01 AE and B, or rather native strains of the two viruses.
Adult ; China ; epidemiology ; Epidemics ; Genetic Variation ; HIV Infections ; diagnosis ; epidemiology ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Homosexuality, Male ; statistics & numerical data ; Humans ; Male ; Middle Aged ; Molecular Epidemiology ; Molecular Sequence Data ; Phylogeny ; Young Adult

OBJECTIVETo explore the effects of different fertilization methods on the outcomes of elective blastocyst culture.

METHODSWe retrospectively analyzed the outcomes of elective blastocyst culture for 1 153 cycles of IVF and 205 cycles of ICSI performed between january 2009 and December 2012.

RESULTSA total number of 14 748 embryos in the IVF group and 2 655 embryos in the ICSI group underwent sequential blastocyst culture, with 7 871 blastocysts formed in the former and 1 210 in the latter. No cycles were canceled for no blastocyst formation in either of the two groups. The rates of quality embryos, blastocyst formation and embryo utilization were significantly higher in the IVF than in the ICSI group (64.77 vs 58.72%, 53.37 vs 45.57%, and 60.06 vs 52.17%, all P < 0.05), but the rates of implantation, clinical pregnancy and abortion showed no significant differences between the two groups (48.94 vs 51.43%, 49.03 vs 52.02%, and 11.69% vs 15.56, all P > 0.05).

CONCLUSIONWith the same inclusion criteria of selective blastocyst culture, IVF has a lower risk of cycle cancellation due to no blastocyst formation and therefore may effect higher rates of blastocyst formation and embryo utilization than ICSI. Our study suggested that appropriate inclusion criteria of selective blastocyst culture should be laid down according to different fertilization methods.

Adult ; Blastocyst ; Embryo Transfer ; Female ; Fertilization in Vitro ; methods ; Humans ; Pregnancy ; Retrospective Studies ; Sperm Injections, Intracytoplasmic

BACKGROUND: Sj?gren's syndrome (SS) is a chronic progressive autoimmune disease. The incidence of peripheral nervous system damage in patients with primary Sj?gren's syndrome (pSS) is 10%-30%. Previous studies have shown that there are multiple electrophysiological manifestations in patients with pSS presenting with peripheral neuropathy. However, there is no consensus on its neuroelectrophysiological manifestations. Peripheral neuropathy associated with pSS is easily confused with peripheral neuropathy caused by other etiologies. OBJECTIVE: To observe the neuroelectrophysiological manifestations of peripheral neuropathy associated with pSS to assist in the diagnosis of the disease. METHODS: A total of 100 pSS patients with peripheral neuropathy who receive treatment in the Department of Neurology, First Affiliated Hospital of Kunming Medical University, in China will be included in this study. Fifty-two patients included in the preliminary experiment presented with peripheral neuropathy associated with pSS. The primary outcome measure is the rate of abnormal motor nerve conduction velocity. The secondary outcome measures include the rate of abnormal terminal motor latency, the rate of abnormal compound muscle action potential amplitude, the rate of sensory nerve conduction velocity, the rate of abnormal sensory nerve action potential amplitude, the rate of abnormal F wave, and the rate of abnormal sympathetic skin response. RESULTS AND CONCLUSION: Results of 52 patients included in the preliminary study showed that the rate of each electrophysiological index was similar between upper and lower extremities; the rate of abnormal motor nerve conduction velocity was significantly higher than the rate of abnormal compound muscle action potential amplitude; the rate of sensory nerve conduction velocity was significantly higher than the rate of abnormal sensory nerve action potential amplitude; the rate of abnormal motor nerve conduction velocity was similar to the rate of abnormal sensory nerve conduction velocity; the rate of abnormal compound muscle action potential amplitude was similar to the rate of abnormal sensory nerve action potential amplitude; the rate of abnormal wave was significantly lower than the rate of abnormal motor nerve conduction velocity; the rate of abnormal sympathetic skin response was similar to the rate of abnormal motor nerve conduction velocity. Results from this study will reveal neuroelectrophysiological abnormality in peripheral neuropathy associated with pSS, which will help diagnose the disease.

To explore the genetic characteristics of viral quasispecies in HIV-1 CRF07_BC infections among intravenous drug users (IDU), the gp120 fragments of HIV-1 env gene were amplified from plasma samples collected from 6 CRF07_BC infected persons using single genome amplification and sequencing (SGA/ SGS) method, and 11 to 28 sequences were obtained from these samples, respectively, A neighbor-joining phylogenetic tree was reconstructed to describe the genetic characteristics of viral quasispecies. The Simplot, segments' phylogenetic trees and diversity plots based on average pairwise distance (APD) were used to identify the recombination events between quasispecies. The SGA sequences derived from single specimen formed a large monophyletic cluster in the neighbor-joining phylogenetic tree and showed the complex topologic structures of viral quasispecies. Of the 6 CRF07_BC infected patients, only one possessed the high genetic homogeneity, whereas the other five individuals showed high heterogeneity, with two to four subclusters inside the monophyletic cluster for each specimen. In addition, the recombinant events were identified among viral quasispecies from 3 cases. The results show SGA technique and phylogenetic analyses are useful tool to investigate the intrahost CRF07_BC gp120 complex quasispecies variation and high genetic diversity.
Adult ; Drug Users ; Female ; HIV Infections ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Humans ; Male ; Molecular Sequence Data ; Phylogeny ; Substance Abuse, Intravenous ; virology ; Young Adult

OBJECTIVETo analyze the protein expression profiles of multinodular hepatocellular carcinoma (HCC) with multicentric occurrence (MO) or with intrahepatic metastasis (IM).

METHODS5 IM and 6 MO patients were divided into groups of IM1, IM2, MO1 and MO2 according to the size of node of HCC. Two dimensional gel electrophoresis (2-DE) and mass spectrum were used to analyze the protein expression profiles. Western blot was used to confirm the results obtained by mass spectrum.

RESULTS2-DE of IM1, IM2, MO1 and MO2 indicated that 30 protein dots were differentially expressed in these tumors. By mass spectrum, 25 proteins were identified. Gene ontology classification indicated that these proteins are associated to cell movement, signal transduction, oxidoreduction, lipid metabolism, and amino acid metabolism.

CONCLUSIONThe protein expression profiles of IM is different from that of MO, 2-DE and mass spectrum can be used to identify the molecular markers of IM and MO of HCC.

Adult ; Blotting, Western ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasms, Multiple Primary ; metabolism ; pathology ; Prognosis ; Proteome ; metabolism ; Proteomics

Objective To survey avian influenza A viruses (AlVs) in the environment and explore the reasons for the surge in human H7N9 cases.Methods A total of 1,045 samples were collected from routine surveillance on poultry-related environments and 307 samples from human H7N9 cases-exposed environments in Henan from 2016 to 2017.The nucleic acids of influenza A (Flu A),H5,H7,and H9 subtypes were detected by real-time polymerase chain reaction.Results A total of 27 H7N9 cases were confirmed in Henan from 2016 to 2017,24 had a history of live poultry exposure,and 15 had H7N9 virus detected in the related live poultry markets (LPMs).About 96％ (264/275) Flu A positive-environmental samples were from LPMs.H9 was the main AIV subtype (10.05％) from routine surveillance sites with only 1 H7-positive sample,whereas 21.17％ samples were H7-positive in H7N9 cases-exposed environments.Samples from H7N9 cases-exposed LPMs (47.56％)had much higher AIVs positive rates than those from routine surveillance sites (12.34％).The H7+H9 combination of mixed infection was 78.18％ (43/55) of H7-positive samples and 41.34％ (43/104) of H9-positive samples.Conclusion The contamination status of AIVs in poultry-related environments is closely associated with the incidence of human infection caused by AlVs.Therefore,systematic surveillance of AlVs in LPMs in China is essential for the detection of novel reassortant viruses and their potential for interspecies transmission.

Purpose::Vibrio vulnificus ( V. Vulnificus) infection is characterized by rapid onset, aggressive progression, and challenging treatment. Bacterial resistance poses a significant challenge for clinical anti-infection treatment and is thus the subject of research. Enhancing host infection tolerance represents a novel infection prevention strategy to improve patient survival. Our team initially identified cytochrome P4501A1 (CYP1A1) as an important target owing to its negative modulation of the body's infection tolerance. This study explored the superior effects of the CYP1A1 inhibitor bergamottin compared to antibiotic combination therapy on the survival of mice infected with multidrug-resistant V. Vulnificus and the protection of their vital organs. Methods::An increasing concentration gradient method was used to induce multidrug-resistant V. Vulnificus development. We established a lethal infection model in C57BL/6J male mice and evaluated the effect of bergamottin on mouse survival. A mild infection model was established in C57BL/6J male mice, and the serum levels of creatinine, urea nitrogen, aspartate aminotransferase, and alanine aminotransferase were determined using enzyme-linked immunosorbent assay to evaluate the effect of bergamottin on liver and kidney function. The morphological changes induced in the presence of bergamottin in mouse organs were evaluated by hematoxylin and eosin staining of liver and kidney tissues. The bacterial growth curve and organ load determination were used to evaluate whether bergamottin has a direct antibacterial effect on multidrug-resistant V. Vulnificus. Quantification of inflammatory factors in serum by enzyme-linked immunosorbent assay and the expression levels of inflammatory factors in liver and kidney tissues by real-time quantitative polymerase chain reaction were performed to evaluate the effect of bergamottin on inflammatory factor levels. Western blot analysis of IκBα, phosphorylated IκBα, p65, and phosphorylated p65 protein expression in liver and kidney tissues and in human hepatocellular carcinomas-2 and human kidney-2 cell lines was used to evaluate the effect of bergamottin on the nuclear factor kappa-B signaling pathway. One-way ANOVA and Kaplan-Meier analysis were used for statistical analysis. Results::In mice infected with multidrug-resistant V. Vulnificus, bergamottin prolonged survival ( p = 0.014), reduced the serum creatinine ( p = 0.002), urea nitrogen ( p = 0.030), aspartate aminotransferase ( p = 0.029), and alanine aminotransferase ( p = 0.003) levels, and protected the cellular morphology of liver and kidney tissues. Bergamottin inhibited interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α expression in serum (IL-1β: p = 0.010, IL-6: p = 0.029, TNF-α: p = 0.025) and inhibited the protein expression of the inflammatory factors IL-1β, IL-6, TNF-α in liver (IL-1β: p = 0.010, IL-6: p = 0.011, TNF-α: p = 0.037) and kidney (IL-1β: p = 0.016, IL-6: p = 0.011, TNF-α: p = 0.008) tissues. Bergamottin did not affect the proliferation of multidrug-resistant V. Vulnificus or the bacterial load in the mouse peritoneal lavage fluid ( p = 0.225), liver ( p = 0.186), or kidney ( p = 0.637). Conclusion::Bergamottin enhances the tolerance of mice to multidrug-resistant V. Vulnificus infection. This study can serve as a reference and guide the development of novel clinical treatment strategies for V. Vulnificus.

Aim To investigate the protective effect of Jujing formula on retina of mice with dry age-related macular degeneration(AMD).Methods The mouse model of dry AMD was induced by intraperitoneal in-jection of sodium iodate,and the prognosis was given to the Jujing formula.Retinal thickness was detected by optical coherence tomography(OCT),the retinal morphological changes were observed by hematoxylin-eosin(HE)staining,and the apoptosis of retinal cells was detected by in situ terminal transferase labeling(TUNEL)staining.Combination of tumor necrosis fac-tor-α(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1 β)in eyeballs and serum,superoxide dis-mutase(SOD),glutathione(GSH)and malondialde-hyde(MDA)were evaluated to assess the protective effects of Jujing formula on retinal injury in mice with dry AMD.Results The results of OCT,HE and TUNEL staining showed that Jujing formula significant-ly improved the retinal injury induced by sodium iodate in mice with dry AMD,increased the retinal thickness(P＜0.05),reduced the apoptosis of retinal cells(P＜0.01),and increased the levels of GSH,IL-6 and SOD activity in eyeballs and serum(P＜0.01).The levels of TNF-α,IL-6,IL-1β and MDA were reduced(P＜0.01).Conclusions Jujing formula has certain therapeutic effects on retinal injury in dry AMD,which may be related to inhibiting inflammatory response and enhancing antioxidant capacity.