Epigenetics change has been considered to be the most promising new strategy for disease control and prevention. TCM regulates gene expression through epigenetics, participating in pathological and physiological process including cell apoptosis, proliferation, differentiation, cell cycle regulation, immunity, inflammation, and metabolism. This article reviewed the application of DNA methylation, histone modification and the miRNA regulation in TCM research.

To obtain the tervalent fusion toxin gene (named FT),three toxin gene fragments from three species of Vibrio parahaemolyticus,Vibrio vulnificus and Vibrio mimicus were connected with the flexible linker (GGGGS) using overla Pextension PCR. The three toxin gene fragments respectively encode the mature proteins of the thermostable direct hemolysin (TDH) of V. parahaemolyticus,the cytotoxin (VVC) of V. vulnificus and the heat-labile hemolysin (VMH) of V. mimicus. The identity of FT nucleic acid sequence was 99.6% with the corresponding toxin gene fragments. The open reading frame of FT was 3225 bp,encoding 1074 amino acid residues with the predicted molecular weight (MW) of 120.4 kDa. Then,FT was subcloned into the expression vector pET-22b(+). The construction of recombinant expression vector pET-22b-FT was followed by transforming into E. coli BL21(DE3) for expression. The SDS-PAGE electrophoresis results indicated that the MW of the fusion toxin protein was matched to the predicted MW. After induction by 1 mmol/L IPTG at 37℃,the fusion toxin protein was effectively expressed in E. coli BL21(DE3) with the amount of 11.49% through thin layer chromatography scanning (TLCS) analysis. Cavia cobaya was immunized using the purified cytorrhyctes to produce the anti-serum. Through the determination of the optimum working conditions,the sensitivity test,the specificity test,repeatability test and sample simulation test,the indirect ELISA method was established,which is a broad-spectrum,rapid and specific to detect various of food-poisoning Vibrio simultaneously.

Objective To observe the effect of Proanthocyanidin on motor after spinal cord injury (SCI) in rats. Methods 36 healthy adult SD rats were divided into groups A, B and C (n=12), and SCI was induced with Allen's mode (250 g·mm) on T9. Proanthocyanidin 40 mg/kg was injected intraperitoneally for group A, methylprednisolone (MP) 30 mg/kg for group B and the same volume of saline for group C 30 min after SCI. 1 d, 3 d and 7 d after operation, all the rats were assessed with Basso-Beattie-Bresnahan (BBB) scale and slanting board test, and their serumal malondialdehyde (MDA) and superoxide dismutase (SOD) were detected. Results The scores of BBB and the slanting board test imporoved more in group A and group B than in group C (P<0.05). The SOD increased and MDA decreased in groups A and B significantly 1 d and 3 d after operation compared with those of group C (P<0.05), and only in group A 7 d after operation (P<0.05). Conclusion Proanthocyanidin may inhibit the lipid peroxidation and promote the recovery of motor after spinal cord injury in rats.

Objective To investigate the preoperative magnetic resonance imaging (MRI)examination of the distribution of neurovascular bundles (NVB) around the prostatic capsule,and its clinical value in the nerve-sparing laparoscopic radical prostatectomy (NS-LRP).Methods The clinical data of 42 patients with clinically localized prostate cancer who were admitted from January 2008 to January 2017 were retrospectively analyzed.Age ranged from 58 to 74 years,with an average of 68 years.Preoperative serum PSA range from 0.94 to 12.28 ng/dl,with an average of 7.01 ng/dl.Preoperative Gleason score range from 6 to 8,with an average of 6.Clinical stage:T1-T2 37 cases,T3 5 cases.The average preoperative International Erectile Function Index questionnaire-5 (IIEF-5) 21,of which 23 cases had normal erectile function (IIEF-5 ＞ 22).All the 42 patients underwent MRI examination before operation.According to the distribution of NVB around the prostatic capsule,they were divided into 3 groups:17 cases in group A,and no NVB was evident in all cases.In group B,8 cases were visible but not obvious.In group C,17 cases were evident NVB.There was no significant difference in age,preoperative serum PSA and Gleason score between the three groups (P ＞ 0.05).The preoperative IIEF-5 in group A,B,and C were 19.5,22.8,and 21.5,respectively,with no statistically significant difference (P ＞ 0.05).All 42 cases received NS-LRP under general anesthesia.The differences in IIEF-5 before and after surgery were compared between the three groups.Results In this study,42 cases were successfully completed.42 patients were followed up for 12 to 36 months,with an average of 14.1 months.In group A,B,and C,postoperative IIEF-5 was 8.0,14.1,and 15.5,respectively,which was statistically significant compared with preoperative values (P ＜ 0.05).The decrease of IIEF-5 afteroperation in group A was significantly different from that of group Band C (P ＜0.05).Conclusions Compared with patients with visible NVB on MRI examination,patients have no visible NVB observed on MRI with erectile function-related nerves around the prostatic capsule may be walking on both sides of the prostatic capsule and spreading over the entire anterior ventral surface of the prostate.To maxium-preserve NVB and postoperative erectile function to the utmost,NS-LRP surgery should be more accurately dissected on both sides and ventral side of the prostatic capsule.

Objective To evaluate the safety and efficacy of ultrasound-guided transperitoneal PCNL for treatment of renal calculi in pelvic ectopic kidney.Methods The clinical data of 4 cases of pelvic ectopic kidney treated in our hospital from June 2016 to August 2017 were retrospectively analyzed.The patient's age ranged from 30 to 67 years with an average of 47.25 years.There were two male and two female patients.2 cases suffered with left pelvic ectopic kidney stones.One case had right pelvic horseshoe ectopic kidney stones and one case had spinal deformity left pelvic ectopic kidney stones.Multiple stones were found in 2 cases.The mainly stones located in the renal pelvis in one case.Multiple stones in the lower calyx was found in another case.The remaining 2 cases had single stones in the renal pelvis.Stone size was 3.4 cm × 2.1 cm,3.0cm×2.1 cm,2.8 cm×2.4 cm and 2.1 cm×1.9 cm respectively.And 1 case with renal calculi in left pelvic ectopic kidney had a history of open surgerv 11 years ago because of the same situation with pelvic ectopic kidney calculi.In this study,all operation was performed under general anesthesia.The ultrasound-guided percutaneous nephrolithotomy combined with preoperative CTU and intraoperative color Doppler ultrasound was used to find the target renal calyx or renal pelvis.And a certain pressure was applied to the abdominal wall to make the intestinal canal deviate from the surface of the pelvic ectopic kidney as much as possible.Under ultrasound real-time detection,it was punctured into the target renal calyx or renal pelvis to find and break stones.And F6 double J tube and F18 nephrostomy tube were left.Operative and post-operative related details (stone free,bleeding,abdominal infection) were reviewed.Results Stone free rate were 100％ (4/4).The time of operation and hospital stay were 62,59,55,51 min and 6,6,5,5 days,respectively.None had significant intraoperative and postoperative complications,including haemorrhage,injury of mesenteric vessels,abdominal infection and hydrops.No one left residual stones during 1 month and 3 nonths of follow-up.Conclusions The ultrasound-guided transperitoneal PCNL is a safe and effective procedure for the treatment of patients with renal calculi in pelvic ectopic kidney.

OBJECTIVETo investigate the association of interleukin 6 gene (IL-6) promoter region 634C/G (rs1800796) single nucleotide polymorphism (SNP) with the genetic susceptibility to endometriosis (Ems) in south Han Chinese women.

METHODSA case-control study was performed in 432 Ems patients and 499 control women to evaluate the SNP of IL-6 634C/G by using a fluorescent quantitative PCR-based high resolution melting (HRM) method.

RESULTSThere were statistical significances in the IL-6 634C/G alleles, whether or not to carry allele G and genotype distributions between Ems patients and control women (P=0.032, 0.014 and 0.045, respectively). Allele C enhanced the risk of Ems 1.057 times while allele G reduced the risk of Ems 0.835 time. Carrying allele G reduced the risk of Ems 0.822 time, whereas not carrying allele G enhanced the risk of Ems 1.143 times. Compared with genotype CC, the risk of Ems with genotype CG reduced 0.704 time (95% CI: 0.533-0.931). There was no significant difference in whether or not carrying allele G distribution between Ems patients and control women (P=0.729).

CONCLUSIONThe present study demonstrated significant association between the SNP of IL-6 634C/G and genetic susceptibility to Ems in south Han Chinese women.

Alleles ; Case-Control Studies ; Endometriosis ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Interleukin-6 ; genetics ; Polymorphism, Single Nucleotide ; genetics

Objective: To evaluate efficacy and safety of retrograde intrarenal stone surgery for treatment of upper urinary calculi. Methods: The clinical data of 640 patients with upper urinary tract calculi treated by retrograde intrarenal stone surgery (RIRS) in Renmin Hospital of Wuhan University from April 2016 to January 2019 were retrospectively analyzed. There were 424 males and 216 females. The awerage age was (46.2±12.8) years old, ranging 18 to 76 years old. The maximum diameter of the stone is (1.4±0.7) cm, ranging 0.6-3.2 cm. There were 126 cases with inferior calculi and 514 cases with non-lurgical calculi. There were 196 cases with unilateral ureteral calculi, 118 unilateral ureteral calculi cases with renal calculi, 236 cases with unilateral renal stones, and 90 cases with double kidney stones. 104 cases were placed with double J tube before operation and 496 cases were not placed before operation. There were 8 cases of horseshoe kidney, 30 cases of isolated kidney with renal insufficiency, 4 cases of pelvic ectopic kidney with dysplasia, 6 cases of congenital ureteral malformation and 2 cases of sponge kidney. Preoperative average hemoglobin was (133.2±5.6)g/L, ranging 126-188 g/L.And average serum creatinine was (84.4±12.2)μmol/L, ranging 74-242μmol/L before operation. All patients were treated with general anesthesia under the lithotomy position. The ureteroscopy combined with holmium laser lithotripsy was performed.The 200μm fiber was used, which the parameters were set as 12-45 W(0.5-1.5 J/10-30 Hz). The stone baskets were used to take stones according to actual conditions. The operation was performed by doctors of the same qualifications. Results: All patients underwent successful operation. The mean operation time was (45.6±14.6)min. The average postoperative hospitalization was (4.8±1.5)d. The postoperative serious complication rate was 0.9%, including(2 cases of sepsis and 1 case of subcapsular hematoma. Of the 640 patients, 596 were admitted to the hospital for a double J tube and 44 were lost of follow-up. 552 patients met the stone removal criteria, 44 patients did not meet the stone removal criteria for other treatments, such as extracorporeal shock wave lithotripsy, ureteroscopy or observed regularly. The stone-free rate (SFR) was 92.6% (552/596) after 1-3 months. On the first postoperative day, serum creatinine was (76.0±10.6)(58-156) μmol/L, and postoperative hemoglobin was (126.4±9. 6)(120-176) g/L. There was no significant difference in preoperative and postoperative hemoglobin (t=2.02, P=0.064). Preoperative and postoperative creatinine (t=64.76, P<0.05) was statistically significant. Meanwhile, the stone size (χ²=29.569, P<0.05) and position (χ²=44.949, P<0.05) versus SFR the impact was statistically significant. Multivariate regression analysis showed that stone size was not an independent risk factor for stone clearance (P=0.639). The stone position was an independent risk factor for stone clearance (P=0.013). Conclusions RIRS is a reliable treatment for small and medium calculi patients of the upper urinary tract. The curative effect of stone removal is clear, the complications are few, the safety is high. However, there are certain limitations to the efficacy in the treatment of large stones and lower calculi. Lower calculi is the independent risk factor for the treatment of efficacy.

Objective To explore the clinical significance of fused renal pyramid (FRP) structure in the establishment of percutaneous renal access.Methods From May 2017 to April 2018,10 fresh porcine kidneys were selected to cast in blood vessels for grading the kidney artery.Then another 80 isolated porcine kidueys were used to simulate percutaneous renal pu ncture and dilatation to establish F24 operative access by the same surgeon.Under the endoscope and microscope,we compared the effects of four different puncture paths on the occurrence of renal vascular injury when respectively punctured through the normal renal pyramid (group A),the side of the FRP (group B),the centre of the FRP (group C) and the renal column (group D).Results The kidney arteries can be divided into six grades,there is grade Ⅳ branchinterlobar artery walking inside the FRP.The diameter of interlobar artery in the FRP was significantly smaller than that in the renal column (0.442 ±0.012) mm vs.(0.778 ±0.037) mm,(P ＜0.001).Endoscopic observation and pathological tissue section showed the following results.In group A,there was no injured blood vessel distributed along the access.There were six specimens with grade Ⅴ or Ⅵ arteries injury in the cortex.Owing to the small size of the renal pyramid and the inaccurate location of the puncture,there was also injury associated with a normal grade Ⅳ artery in the renal column.In group B,there was a certain distance between the tract and the grade Ⅳ artery that distributed in the FRP,injury was still noticed in four specimens.And six specimens have grade Ⅴ/Ⅵ arterial injury.As the distance between the tract and the renal column decreased,there was a case in which a simultaneously injury occurred to the extremity of a grade Ⅲ artery and a grade Ⅳ artery.In group C,there was a white thin strip of connective tissue exposed along the puncture tract.Ectopic grade Ⅳ artery injury occurred in fourteen specimens,and grade Ⅴ/Ⅵ artery injury occurred in seven specimens.In group D,there were grade Ⅲ to Ⅵ arteries distributed along the operational access,which was cowered with white fat and connective tissue.The number of arteryinjury in grades Ⅲ,Ⅳ,and Ⅴ/Ⅵ were4,19,and 5,respectively.The mean ranks of artery injury degree in groups A (17.0),B (30.1),C (33.5) and D (41.5) gradually increased,and the difference was significant (P =0.006).There was a significant difference between group A and C (P =0.018),while no significant difference between group A and B (P =0.122),groups C and D (P =0.072).The proportion of grade Ⅳ artery injury in group A,B,and C was 5％ (1/20),25％ (5/20),and 70％ (14/20),respectively.There was a significant difference between group A and C (P =0.029),while no significant difference between group A and B (P =0.316).There was no significant difference in the injury of grade Ⅴ and Ⅵ artery in four groups (P =0.827).Conclusions When establishing a percutaneous renal access,vascular injury caused bv puncturing through the FRP cannot be ignored.It is necessary to carefu lly identify and bypass the FRP when selecting the puncture path.If unavoidable,the puncture path shoull be on the centreline of one side pyramid of the FRP.

OBJECTIVETo investigate the association of single nucleotide polymorphisms in cytochrome P450 17 (CYP17) and estrogen receptor alpha (ERα ) genes with the risk of endometriosis among southern Chinese women.

METHODSTwo SNPs rs743572 (CYP17 gene 34T/C) and rs9322331 (ERα gene -397T/C) were genotyped by high resolution melting curve in 432 endometriosis patients and 499 matched controls.

RESULTSThere was no significant difference in the genotype frequencies of the two loci between endometriosis patients and the control subjects (P> 0.05). And there was no significant interaction effect of these two genes on the disease either.

CONCLUSIONCYP17 gene and ERα gene may not be genetic risk factors for endometriosis among southern women in China.

Asian Continental Ancestry Group ; genetics ; Endometriosis ; genetics ; Estrogen Receptor alpha ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Single Nucleotide ; Risk Factors ; Steroid 17-alpha-Hydroxylase ; genetics

OBJECTIVETo explore the association between the arylhydrocarbon receptor gene (AhR) 1661G/A or arylhydrocarbon nuclear translocatorgene (ARNT) 567G/C polymorphism and endometriosis in southern Han Chinese women.

METHODSThe polymorphisms of AhR gene 1661G/Aand ARNT gene 567G/C in 431 cases of endometriosis and 499 healthy women were genotyped by fluorescence quantitative PCR-based high resolution melting.

RESULTSThe frequencies of genotypes AA, AG, GG and alleles A and G in controls were 12.0%, 41.9%, 46.1%, 33.0% and 67.0%, respectively, which were not significantly different from those in patients with endometriosis (9.7%, 44.6%, 45.7%, 32.0% and 68.0%, respectively). The genotype frequencies of GG, GC, CC and alleles C and G in controls (15.6 %, 51.7%, 32.7%, 58.5%, 41.5%) were not significantly different from those in patients with endometriosis (13.5%, 47.8%, 38.7%, 62.6%, 37.4%), either. And no interaction of AhR 1661G/A and ARNT 567G/C on endometriosis was found.

CONCLUSIONNo association between AhR 1661G/A and ARNT 567G/C genetic polymorphisms and endometriosis was found in the southern Han Chinese women in this study.

Alleles ; Aryl Hydrocarbon Receptor Nuclear Translocator ; genetics ; China ; Endometriosis ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; Receptors, Aryl Hydrocarbon ; genetics