Nonobstructive azoospermia (NOA), one of the most severe types of male infertility, etiology often remains unclear in most cases. Therefore, this study aimed to detect four biallelic detrimental variants (0.5%) in the minichromosome maintenance domain containing 2 ( MCMDC2 ) genes in 768 NOA patients by whole-exome sequencing (WES). Hematoxylin and eosin (H&E) demonstrated that MCMDC2 deleterious variants caused meiotic arrest in three patients (c.1360G>T, c.1956G>T, and c.685C>T) and hypospermatogenesis in one patient (c.94G>T), as further confirmed through immunofluorescence (IF) staining. The single-cell RNA sequencing data indicated that MCMDC2 was substantially expressed during spermatogenesis. The variants were confirmed as deleterious and responsible for patient infertility through bioinformatics and in vitro experimental analyses. The results revealed four MCMDC2 variants related to NOA, which contributes to the current perception of the function of MCMDC2 in male fertility and presents new perspectives on the genetic etiology of NOA.
Humans ; Male ; Azoospermia/genetics* ; Meiosis/genetics* ; Spermatogenesis/genetics* ; Adult ; Exome Sequencing ; Microtubule-Associated Proteins/genetics* ; Alleles ; Infertility, Male/genetics*

Brain Neoplasms/surgery* ; Cyclic AMP Response Element-Binding Protein ; Gene Fusion ; Humans ; Oncogene Proteins, Fusion/genetics* ; RNA-Binding Protein EWS/genetics*

Since the outbreak of 2019-nCoV, the epidemic has developed rapidly and the situation is grim. LANCET figured out that the 2019-nCoV is closely related to "cytokine storm". "Cytokine storm" is an excessive immune response of the body to external stimuli such as viruses and bacteria. As the virus attacking the body, it stimulates the secretion of a large number of inflammatory factors: interleukin(IL), interferon(IFN), C-X-C motif chemokine(CXCL) and so on, which lead to cytokine cascade reaction. With the exudation of inflammatory factors, cytokines increase abnormally in tissues and organs, interfering with the immune system, causing excessive immune response of the body, resulting in diffuse damage of lung cells, pulmonary fibrosis, and multiple organ damage, even death. Arachidonic acid(AA) metabolic pathway is principally used to synthesize inflammatory cytokines, such as monocyte chemotactic protein 1(MCP-1), tumor necrosis factor(TNF), IL, IFN, etc., which is closely related to the occurrence, development and regression of inflammation. Therefore, the inhibition of AA metabolism pathway is benefit for inhibiting the release of inflammatory factors in the body and alleviating the "cytokine storm". Based on the pharmacophore models of the targets on AA metabolic pathway, the traditional Chinese medicine database 2009(TCMD 2009) was screened. The potential herbs were ranked by the number of hit molecules, which were scored by pharmacophore fit value. In the end, we obtained the potential active prescriptions on "cytokine storm" according to the potential herbs in the "National novel coronavirus pneumonia diagnosis and treatment plan(trial version sixth)". The results showed that the hit components with the inhibitory effect on AA were magnolignan Ⅰ, lonicerin and physcion-8-O-β-D-glucopy-ranoside, which mostly extracted from Magnoliae Officinalis Cortex, Zingiberis Rhizoma Recens, Lonicerae Japonicae Flos, Rhei Radix et Rhizoma, Salviae Miltiorrhizae Radix et Rhizoma, Scutellariae Radix, Gardeniae Fructus, Ginseng Radix et Rhizoma, Arctii Fructus, Dryopteridis Crassirhizomatis Rhizoma, Paeoniaeradix Rubra, Dioscoreae Rhizoma. Finally the anti-2019-nCoV prescriptions were analyzed to obtain the potential active prescriptions on AA metabolic pathway, Huoxiang Zhengqi Capsules, Jinhua Qinggan Granules, Lianhua Qingwen Capsules, Qingfei Paidu Decoction, Xuebijing Injection, Reduning Injection and Tanreqing Injection were found that may prevent 2019-nCoV via regulate cytokines. This study intends to provide reference for clinical use of traditional Chinese medicine to resist new coronavirus.
Arachidonic Acid/metabolism* ; Betacoronavirus ; COVID-19 ; Coronavirus Infections/immunology* ; Cytokines/immunology* ; Drugs, Chinese Herbal/pharmacology* ; Humans ; Medicine, Chinese Traditional ; Metabolic Networks and Pathways ; Pandemics ; Pneumonia, Viral/immunology* ; SARS-CoV-2 ; COVID-19 Drug Treatment

Porous carbon nanoparticles ( NPC) were prepared by ZnCl2 activation and carbonization using citrus waste as carbon source. A sample pretreatment method with NPC as dispersive solid phase extraction (d-SPE ) absorbent was established for the determination of organophosphorus pesticides in fruits and vegetables by gas chromatography. The NPC was characterized by scanning electron microscopy (SEM), X-ray diffraction ( XRD), FT-IR spectra, Raman spectroscopy, Brunauer, Emmett and Teller surface area(BET). Those results showed that the NPC was an amorphous porous carbon material with pore size in the range of 0-15 nm. Its specific surface area and pore volume were 1243 m2 / g and 1. 28 cm3 / g, respectively. The analysis conditions, including the amount and clean up time of adsorbent, were optimized by analysis of 14 kinds of oranophosphorus pesticides in fruits and vegetables with gas chromatography-flame photometric determination(GC-FPD). Moreover, the comparison for NPC with commercial materials of PSA, C18 and GCB was investigated in this study. The results indicated that the purification time was only 2 min using 0. 01 g NPC. The cost of NPC was about 25% of C18 , 21% of PSA and 16% of GCB. Because of the porous structure of NPC, the purification efficiency was significantly higher than the three commercial materials mentioned above. Under the optimum conditions, the calibration curves of the 14 organophosphorus pesticides were linear in the range of 0. 02-1. 00 mg / L with good correlation coefficients (R2>0. 99) and detection limits (S / N=3) of 0. 63-5. 30 μg / kg. The recoveries of the pesticides at three spiked levels ranged from 71. 3% to 114. 7%with the relative standard deviations (RSDs) of 0. 9% -12. 9% . The method is simple, rapid, sensitive, and low cost, and can satisfy the requirements of detection of organophosphorus pesticide residues in fruits and vegetables, displaying a good application prospect.

OBJECTIVETo explore the effect of Wnt signaling suppression on proliferation of non small cell lung cancer to gefitinib, and its related mechanisms.

METHODSPC9 and PC9/AB2 cells of both gefitinib sensitive and resistant were treated with different concentrations of gefitinib, and the proliferation index was measured using CCK8 kit. The members of Wnt signaling pathway were detected by Western blot. Dual luciferase reportor gene assay (TOP Flash) was used to document the transcriptional level of β-catenin. β-catenin siRNA was transfected into PC9/AB2 cells to suppress the Wnt signaling transcription, followed by treatment with different concentrations of gefitinib. Western blot was then used to detect the expression of EGFR and its downstream signaling after inhibit the expression of β-catenin.

RESULTSTreating with different concentrations of gefitinib, the resistance of PC9/AB2 cells to gefitinib was significantly increased (P < 0.05). The members of Wnt signaling expressed at higher level in PC9/AB2 cells than in PC9 cells (t = 24.590, P = 0.000). TOP Flash examination showed that the endogenous transcriptional activity of Wnt signaling was higher in PC9/AB2 cell than that in PC9 cell (t = 4.983, P = 0.008). Compared with the negative control group, apoptotic rate and sensitivity to gefitinib significantly increased in interfered group (P < 0.05). The expression of p-ERK1/2 significantly decreased after Wnt signaling suppression, although other proteins showed no significant alterations.

CONCLUSIONSuppressing the activity of Wnt signaling can partly reverse the celluar resistance to gefitinib in non small cell lung cancer.

Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Quinazolines ; administration & dosage ; pharmacology ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

Objective To determine the incidence and risk factors of HIV infection among heroin addicts receiving methadone maintenance treatment(MMT)in Dehong prefecture,Yunnan province.Methods All heroin addicts who were HIV negative at the initiation of MMT in June 2005 and through June 2011,in Dehong prefecture were included in the cohort analysis.HIV incidence was calculated and related risk factors determined by using Cox proportional hazard regression model.Results A total of 3154 MMT clinic attendants were qualified for this cohort study.By June 2011,1023(32.4％)of them had never received any follow-up HIV testing so were thus referred as loss to follow-up.The other 2131(67.6％)members had received at least one follow-up HIV testing and were observed for a total of 4615.86 person-years.During the period,22 new HIV infections or seroconverters were identified,making the overall HIV incidence as 0.48/100 person-years.The HIV incidence was higher among those who were unemployed,never married,self-reported being injecting drug users(IDUs)and HCV positive at entry into the MMT program.None of those who were always negative on follow-up-urine-testing of morphine was discovered as HIV newly infected during the follow-up period.Data from multiple regression analysis under Cox proportional hazard model indicated that after controlling for confounding variables,non-IDUs at the entry point for the MMT program,were less likely to be HIV newly-infected or seroconverted than IDUs(HR=0.29,95％CI:0.11-0.76).Conclusion MMT prograqm in Dehong prefecture was demonstrated to be fairly effective in reducing HIV transmission through drug use.Those HIV negative attendants at the MMT clinic who were IDUs or keep using drugs during the treatment,were at higher risk of HIV seroconvertion.More efforts were needed to improve the follow-up and HIV testing programs for the MMT clinic attendants.

OBJECTIVETo summarize our experience in adult-to-infant living donor liver transplantation (A-ILDLT) and to analyze the efficacy and complications of A-ILDLT.

METHODSThe clinical data, surgical strategies and complications of 28 adult donors and infantile recipients who underwent A-ILDLT from April 2006 to December 2009 were retrospectively analyzed. These 28 patients (14 boys and 14 girls) aged from 80 days to 11.5 months with body weights of 3.08 to 10.3 kg at the time of operation . They suffered from biliary atresia with decompensated cirrhosis. The living donors were 15 mothers, 9 fathers, 3 grandma and 1 elder brother with ABO compatible with the infantile recipients. 27 Donor organs were the left lateral lobe grafts (segment II, III) and 1 graft was segment II. All patients were followed up for 5 to 24 months.

RESULTSThese grafts were orthotopically transplanted into the infantile recipients. The average length of stay was 9.3 days for the donor group without any complications. Postoperative immunosuppression included prednisone, Cyclosporin and mycophenolate mofetil (MMF). A total of 24 postoperative complications occurred in 20 recipients, including 5 vascular complications, 4 bleeding, 7 pneumonia, 2 bowel obstruction, 4 intestinal perforation and 3 rejection. Three recipients died of hepatic arterial thrombosis (HAT). The perioperative mortality rate of recipients was 10.7% (3/28) and the survival rate was 89.3% in peroperative period. One died of stricture of hepatic vein and 1 of accidental asphyxia during follow-up term. At present, 23 cases are still alive.

CONCLUSIONA-ILDLT has become an effective method to infants with end-stage liver disease. The postoperative vascular complication is the predominant cause of death.

Female ; Humans ; Infant ; Liver Diseases ; surgery ; Liver Transplantation ; methods ; Living Donors ; Male ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo explore the experiences of aortic valve preservation and root reconstruction in patients with Marfan syndrome.

METHODSFrom July 2003 to Dec 2007, 22 patients with Marfan syndrome were treated by aortic valve preservation and root reconstruction. There were 12 male and 10 female, the age ranged from 10 to 57 years old with a mean of (28 +/- 10) years. The operation procedures included reimplantation technique in 9 patients, remodeling technique in 8 patients, and patch technique in 2 patients. In addition, reimplantation technique + total aorta replacement in 1 patient, remodeling technique + "aortic arch replacement + stent-elephant trunk" in 1 patient, patch technique + "aortic arch replacement + stent-elephant trunk" in 1 patient. The patients were followed-up by 17 to 64 months with a mean of (46 +/- 16) months.

RESULTSNo in-hospital and follow-up period death occurred. There was one reexploration for bleeding 1 d postoperative. No valve-related complication occurred during the follow-up. At the end of follow-up, no aortic regurgitation was demonstrate in 16 patients, but mild regurgitation in 4 patients, moderate regurgitation in 1 patient and severe regurgitation in 1 patient. Two patients with moderate and severe aortic regurgitation need reoperation 1 year postoperative.

CONCLUSIONThe early and mid-term results of aortic valve preservation and root reconstruction operations in Marfan syndrome were favorable.

Adolescent ; Adult ; Aorta ; surgery ; Aortic Aneurysm ; etiology ; surgery ; Aortic Valve ; surgery ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Marfan Syndrome ; complications ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo study the different features of hyperplasia in castrated and uncastrated mice after testosterone (T) treatment.

METHODSForty-eight BALB/c mice were randomly divided into 6 groups of 8 in each: castrated (A), uncastrated (B) , castrated + low T (C), uncastrated + low T (D), castrated + high T (E), uncastrated + high T (F). Groups C and D were treated with testosterone solution at the dose of 12.5 mg/(kg d) and Groups E and F at 125 mg/(kg d) for 20 consecutive days, while Groups A and B received saline only. All the mice were sacrificed on the 21st day, their ventral and dorsal prostate glands weighed and their pathological features studied.

RESULTSAtrophic prostates were observed in Group A, but normal in Group B; prostatic hyperplasia was found in both Group C and D, but more obvious in the latter (P <0.05); and a slightly higher degree of hyperplasia was noted in Groups E and F than in C and D. There was an increase in serum T and vascular endothelial growth factor (VEGF) concentration and a decrease in serum estrogen (E2) concentration in the testosterone treated groups.

CONCLUSIONBoth castrated and uncastrated mice develop prostate hyperplasia after short-term testosterone treatment, although in different degrees and with different features, which may help further the studies on the association of castration and androgen with prostate diseases.

Animals ; Hyperplasia ; Male ; Mice ; Mice, Inbred BALB C ; Orchiectomy ; Prostate ; pathology ; Prostatic Hyperplasia ; drug therapy ; pathology ; Testosterone ; therapeutic use

Objective Throush identify biochemical characteristics and virulence factors of 2 strains suspected Yersinia pestis(Y.pestis)isolated from the dead Marmota himalayana(M.himalayana)to confirm the nature epidemic focus in Dege County,Sichuan Province.Methods Y.pestis was analyzed by specific staining and shape,culturing characteristics,splitting-test by bacteriophage,test of biochemical characteristics and glycolysis ability,virulence factors,virulence,nutritional requirement,plasmid,genetic test and genetic type. Results The tested strains were Gram staining bacilus.The main biochemical characteristics were Arabinose(+)、 Rhamnose(-),Maltose(+),Melibiose(-),Glycerol(+),Denitrification(+).The virulence factors with FI+.VW+, Pgm+,Pst I+;and with the common 6.0×106,45.0×106,65.0×106 plasmids,also with the virulence-relative plasmid gene.Both their absolutely lethal dose(LD100)in mice were 50 bacteria.The nutritional requirement appeared which were depended on Phenylalanine and Methionine.With the Genomovar 5 genotype characteristics of M.himalayana plague foci of Qinghai-Tibet plateau.The difference between tested strains and Yersinia pseudotubercuosis on the 3 different culture medium was obvious.The tested strains had a Y.pestis' specific 3a fragment,Pst I and FI-Ag,at 22 ℃,the strains could be split by bacteriophage completely.Conclusions According to the diagnostic criteria of plague in China,the 2 suspected strains isolated from Dege County,Sichuan Province ale confirmed as Y.pestis.both with powerful virulenceand with the characteristics of the Y.pestis of M.himahtyana in Qinghai-Tibet plateau plague natural focus.