Male infertility is a multifactorial disease, of which the cause of male infertility cannot be determined, which is called idiopathic male infertility, and the incidence rate is gradually rising. Because its cause is unknown, it has become a major problem in the department of reproductive endocrinology. With the in-depth study of epigenetics, the diagnosis and treatment of idiopathic male infertility also has a new direction, especially the important role of DNA methylation in spermatogenesis and embryonic development. More and more gene fragments and loci have been found by scholars, which makes it possible to achieve accurate identification and targeted treatment. This article reviews and summarizes the research progress of DNA methylation related to idiopathic male infertility in recent years, aiming to further elaborate the pathogenesis of idiopathic male infertility and provide new ideas for clinical diagnosis and treatment.

Ketone bodies have beneficial metabolic activities, and the induction of plasma ketone bodies is a health promotion strategy. Dietary supplementation of sodium butyrate (SB) is an effective approach in the induction of plasma ketone bodies. However, the cellular and molecular mechanisms are unknown. In this study, SB was found to enhance the catalytic activity of 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), a rate-limiting enzyme in ketogenesis, to promote ketone body production in hepatocytes. SB administrated by gavage or intraperitoneal injection significantly induced blood ß-hydroxybutyrate (BHB) in mice. BHB production was induced in the primary hepatocytes by SB. Protein succinylation was altered by SB in the liver tissues with down-regulation in 58 proteins and up-regulation in 26 proteins in the proteomics analysis. However, the alteration was mostly observed in mitochondrial proteins with 41% down- and 65% up-regulation, respectively. Succinylation status of HMGCS2 protein was altered by a reduction at two sites (K221 and K358) without a change in the protein level. The SB effect was significantly reduced by a SIRT5 inhibitor and in Sirt5-KO mice. The data suggests that SB activated HMGCS2 through SIRT5-mediated desuccinylation for ketone body production by the liver. The effect was not associated with an elevation in NAD⁺/NADH ratio according to our metabolomics analysis. The data provide a novel molecular mechanism for SB activity in the induction of ketone body production.
Mice ; Animals ; Butyric Acid/metabolism* ; Ketone Bodies/metabolism* ; Liver/metabolism* ; Hydroxybutyrates/metabolism* ; Down-Regulation ; Sirtuins/metabolism* ; Hydroxymethylglutaryl-CoA Synthase/metabolism*

Sequencing-based spatial transcriptomics(ST)is an emerging technology to study in situ gene expression patterns at the whole-genome scale.Currently,ST data analysis is still complicated by high technical noises and low resolution.In addition to the transcriptomic data,matched histopathological images are usually generated for the same tissue sample along the ST experiment.The matched high-resolution histopathological images provide complementary cellular phenotypi-cal information,providing an opportunity to mitigate the noises in ST data.We present a novel ST data analysis method called transcriptome and histopathological image integrative analysis for ST(TIST),which enables the identification of spatial clusters(SCs)and the enhancement of spatial gene expression patterns by integrative analysis of matched transcriptomic data and images.TIST devises a histopathological feature extraction method based on Markov random field(MRF)to learn the cellular features from histopathological images,and integrates them with the transcrip-tomic data and location information as a network,termed TIST-net.Based on TIST-net,SCs are identified by a random walk-based strategy,and gene expression patterns are enhanced by neighborhood smoothing.We benchmark TIST on both simulated datasets and 32 real samples against several state-of-the-art methods.Results show that TIST is robust to technical noises on multiple analysis tasks for sequencing-based ST data and can find interesting microstructures in dif-ferent biological scenarios.TIST is available at http://lifeome.net/software/tist/and https://ngdc.cncb.ac.cn/biocode/tools/BT007317.

Objective To observe the clinical value of cone-beam computed tomography (CBCT) combined with iGuide system in percutaneous transthoracic needle biopsy (PTNB) of pulmonary lesions.Methods A total of 30 patients with solitary lung lesion underwent PTNB procedures were enrolled.The needle path was planned with iGuide system and 3D CT-like MPR images after CBCT.A coaxial system with 18-gauge cutting needle was used for biopsy.Procedure related data,complications and radiation exposure of patients were recorded.Results Among 30 patients,malignant lesions were found in 24,and benign lesions were in 6 patients.On PTNB,28 patients were correctly diagnosed,1 case was with insufficient samples but obtained correct pathologic result with second biopsy,1 case was false-negative finding.The diagnostic accuracy,sensitivity,specificity,positive predictive value and negative predictive value of PTNB in diagnosis of lung diseases was 96.67％ (29/30),96.00％ (24/25),100％ (5/5),100％ (24/24) and 83.33％ (5/6),respectively.The mean procedure time was (13.03+3.61)min,and exposure dose was (7.95+4.57)mSv.After PTNB procedures,pneumothorax and hemoptysis occurred in 6 (6/30,20.00％) and 3 (3/30,10.00％) patients,respectively.Conclusion CBCT combined with iGuide system for guiding PTNB is accurate and safe with reasonable radiation exposure.

Objective To compare the effects of different doses of dexmedetomidine on postoperative analgesia and sedation.Methods From January 2014 to November 2015,60 cases who needed postoperative analgesia and sedation in our hospital were selected.According to the dosage of dexmedetomidine,they were divided into control group and observation group,30 cases in each group.Two groups of patients were given analgesia pump for analgesia, with 0.5μg·kg-1·h-1,1.0μg·kg-1·h-1 two doses of dexmedetomidine,and analgesia 48 h.At different time points,the VAS pain score,Ramsay sedation score of the two groups were compared,the calm satisfaction in both two groups was recorded,the average incidence of delirium and delirium score in the process of the treatment and the occurrence of adverse reactions were observed.Results With the extension of time of postoperative analgesia,VAS scores in each group decreased,VAS scores in the observation group after 4h,8h,12h were (3.01 ±0.53)points, (1.95 ±0.58)points,(1.52 ±0.35)points,which were lower than those of the control group[(3.92 ±0.32)points, (2.86 ±0.67)points,(2.25 ±0.78)points],the differences were statistically significant (t=3.42,4.11,2.43,all P<0.05).After 24h,48 h,VAS score between the two groups had no significant difference(P>0.05).Postoperative 4h Ramsay score between the two groups had no significant difference (P >0.05 ).With increased postoperative analgesia time,Ramsay scores in two groups decreased,postoperative 8h,12h,Ramsay scores in the observation group [(2.95 ±0.83)points,(2.22 ±0.55)points]were lower than the control group[(3.76 ±0.78)points,(2.98 ± 0.89)points,t=3.45,2.38,all P<0.05].Postoperative 24h,48 h,the Ramsay scores between the two groups had no significant difference(P>0.05).The sedation satisfaction of the observation group (96.67%)was higher than the control group (86.67%),the incidence rate of delirium and delirium average score [3.33%,(15.11 ± 2.03)points]were lower than the control group[13.33%,(19.23 ±2.21 )points],the differences were significant between the two groups (t =4.32,4.32,3.27,all P<0.05 ).After treatment,the adverse reactions were mainly bradycardia,nausea,vomiting,drowsiness,respiratory depression,urinary retention.The overall incidence rate of adverse reactions between the two groups had no significant difference (P >0.05 ).Conclusion The effect of 1 .0μg·kg-1 · h-1 concentration dexmedetomidine within 24h of postoperative analgesia sedative is best,it can reduce the incidence of postoperative delirium,and without obvious drug side effects.

A 49-year-old male patient presented with repeated oral erosions for 1 year,as well as cutaneous erythema and blisters for 1 month.According to histopathological examination and detection of specific antibodies of pemphigus,the patient was diagnosed with pemphigus vulgaris.After the treatment with oral prednisone and azathioprine for 1 month,the white blood cell count and segmented neutrophilic granulocyte count both decreased.After withdrawal of azathioprine,the patient was subcutaneously injected with 150 μg recombinant human granulocyte colony-stimulating factor for 1 session.Then,the white blood cell count became normal.Genotyping test revealed that the patient carried a heterozygous mutation in the NUDT15 gene (JZ274),and was homozygous for wild-type TPMT*2,TPMT*3C and ITPA genes.The patient was diagnosed with azathioprine-induced myelosuppression.

Objective To determine the optimum dose of oxycodone inhibiting responses to tracheal intubation with a double-lumen endobronchial tube in patients undergoing one-lung ventilation (OLV).Methods Sixty adult patients aged 55-64 yr, weighing 60-80 kg, with American Society of Anesthesiologists physical status Ⅰ or Ⅱ, scheduled for elective thoracic surgery requiring OLV, were randomly divided into 3 different doses of oxycodone groups (O1-3 groups, n =20 each).Anesthesia was induced with iv midazolam 0.05 mg/kg, oxycodone 0.30, 0.35 and 0.40 mg/kg (O1.3 groups, respectively) , propofol 1.5 mg/kg and rocuronium 0.9 mg/kg.The patients were tracheally intubated using a double-lumen endobronchial tube and mechanically ventilated.Before anesthesia induction (T0) , immediately before and after intubation (T1,2) , and 1 and 5 min after intubation (T3.4) , arterial blood samples were taken to determine the concentrations of serum norepinephrine (NE) and epinephrine (E) using high-performance liquid chromatography.The occurrence of bucking, body movement, hypertension, and tachycardia were observed.Results The concentrations of serum NE and E were significantly increased at T2,3 than at T1 in group O1 (P＜0.05).Compared with group O1 , the concentrations of serum NE and E were significantly decreased at T2,3 , and the incidence of bucking, body movement, hypertension, and tachycardia was decreased in O2 and O3 groups (P＜0.05).There was no significant difference in the parameters mentioned above between O2 and O3 groups (P ＞ 0.05).Conclusion The optimum dose of oxycodone inhibiting responses to tracheal intubation with a double-lumen endobronchial tube is 0.35 mg/kg in patients undergoing OLV.

OBJECTIVETo analyze the effect of small interfering RNA (siRNA) targeting mouse epididymis-specific colipase-like (meClps) gene on mouse sperm mobility.

METHODSThe eukaryotic expression vector pDsRed2.0-C1-meClps was constructed and transfected into NIH-3T3 cells, and the protein expression was detected with anti-meClps serum. Three interfering sequences targeting meClps (RNAi-251, 224 and 286) were inserted into lentiviral vectors pRNAT-U6.2/lenti, which were co-transfected with pDsRed2.0-C1-meClps into NIH-3T3 cells. The RNA interfering efficiency was confirmed by semi-quantitative PCR and Western blotting. The lentivirus, packed with the lentiviral vector with the highest interfering efficiency, was injected into the caput tissues of mouse epididymis, and its effect on sperm mobility of the cauda epididymis was evaluated.

RESULTSAll the 3 lentiviral RNAi vectors targeting meClps could inhibit the mRNA and protein expressions of meClps, among which pRNAT-U6.2/lenti-RNAi-251 had the highest interfering efficiency. The lentivirus packed with pRNAT-U6.2/lenti-RNAi-251 significantly reduced the path velocity of cauda sperm after injection into the caput epididymis of the mice (P<0.05).

CONCLUSIONKnock-down meClps expression by lentiviral-mediated RNA interference can lower sperm mobility of mice.

Animals ; Epididymis ; Gene Targeting ; Genetic Vectors ; Lentivirus ; Male ; Mice ; NIH 3T3 Cells ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Sperm Motility ; Spermatozoa ; Transfection

Objective To investigate the effect of esmolol pretreatment on propofol injection pain..Methods Ninety patients undergoing breast cancer surgery under general anesthesia were ran-domly assigned into three groups (n=30 each).Group E were pretreated with 5 mg/ml(total 2 ml)es-molol group L with 20 mg/ml (total 2 ml)lidocaine and group N with 2 ml normal saline.After one minute,each group was administrated propofol intravenouly.The pain and hemodynamic data were re-corded.Results Compared with group N,propofol injection pain degree decreased obviously in groups E and L (P <0.05).propofol injection pain occurred in 25 (83.3%)in group N,was signifi-cantly higher than that of 12 (40.0%)in group E and 14 (46.7%)in group L (P <0.05),propofol injection pain had no significant difference between groups E and L.Compared with T1 ,SBP,DBP decreased in groups E and L at T2 ,SBP decreased in group N at T2 significantly (P <0.05).Com-pared with T2 ,DBP was significantly higher at T3 in group E (P <0.05).Conclusion Pretreatment with low dose esmolol was effective in attenuating pain during propofol injection.

Objective To investigate the role of Xuebijing injection in inhibiting perioperative inflammatory responses and protecting the function of multiple organs.Methods A single-blind,randomized,parallel controlled trial was conducted.60 patients in the First Affiliated Hospital of Zhejiang University School of Medicine,aged 18 to 80 years,ASA grade Ⅰ-Ⅲ,undergoing elective abdominal surgery,were enrolled.The patients were randomly divided into the control group (n =30) and the treatment group (n =30).In the control group,after induction of anesthesia,a continuous infusion of 0.9％ normal saline (NS) 200 mL was given in a speed of 2 mL/min,while a continuous infusion of Xuebijing 2 mL/kg in 100 mL of 0.9％ NS was given at 2 mL/min in the treatment group after induction of anesthesia.The blood sample was drawn,and body temperature,routine blood test,C-reactive protein (CRP),liver and kidney function,fasting glucose (Glu),and serum interleukin-6 (IL-6),high mobility group protein B 1 (HMGB 1) levels were determined in all the patients before anesthesia (T1),at the end of operation (T2),12 hours after operation (T3),or at 5:00 am on the third day after operation (T4).At the same time the adverse reactions were recorded for evaluation of the safety of Xuebijing.Results After using Xuebijing injection,T3 body temperature and the T3-T1 temperature difference in treatment group were significantly lower than those of the control group(℃℃:36.70 ± 0.37 vs.37.38 ± 0.47,t＝6.199,P＝0.000; 0.07 ± 0.50 vs.0.85 ±0.58,t＝5.598,P＝0.000).Postoperative white blood cell count,neutrophil percentage,and CRP were significantly higher than those before the operation,but the differences between two groups were not statistically significant.Compared with the control group,alanine aminotransferase (ALT),aspartate transaminase (AST),total bilirubin (TBil) levels at T3 of treatment group were significantly reduced [ALT (U/L):17.56 ± 9.80 vs.88.60 ± 179.76,AST(U/L):27.53 ± 13.12 vs.84.16 ± 151.14,TBil(μ,mol/L):15.46 ± 9.79 vs.25.63 ± 25.33,all P＜0.05].Difference of conjugated bilirubin (CB),blood urea nitrogen (BUN),creatinine (Cr),Glu were not statistically significant between two groups.IL-6 showed an increasing trend after the operation in both groups,and IL-6 level (ng/L) at T2 of the treatment group was significantly lower than that of the control group (41.42 ± 59.74 vs.124.84 ± 119.66,t=3.405,P=0.001).The HMGB 1 level of two groups at T4 were lower than those at T1,but it decreased significantly only in treatment group (μg/L:22.03 ± 15.73 vs.45.09 ± 33.79,P＜0.05),and there was no significant difference between two groups.No serious adverse events occurred during the clinical trial.Conclusions Application of Xuebijing injection during anesthesia can significantly diminish postoperative inflammatory injury,which plays an important role in the protection of liver function,helps restore organ function and improve prognosis,and it is safe and effective.