Objective To establish a novel PCR method that can differentiate the two biotype of Mycoplasma urealytium rapidly based on the disparities of parC gene sequences for clinical routine examination.Methods Design two pairs of specific primers and probes for the targeted gene according to the differences of the parC gene sequences from 14 standard serotypes.The specificity of this amplification was verified by detecting two Mycoplasma urealyticum standard strains including serovar1 and serovar4,50 clinical isolated strains ( 12 are Uu strains and 38 are Up strains) and 7 common bacterias from vagina.Collected 70 swab specimens from patients of Sexually Transmit Department with urogenital inflammation symptom and 71 swab specimens from the gynecology health examination population.All those specimens were detected by culture and our method respectively.The sensitivity of this method was evaluated by comparing with the culture.Differences of the infection rate and distribution of biotypes between different populations were analyzed using statistical software.Results Standard strains of Mycoplasma urealyticum and clinical isolates can be typed into two species by the PCR method without nonspecific amplification.The sensitivity of PCR method is much higher than that of culture ( P＜0.05).The infection rates of Uu,Up and the mixing were 8.57％,61.40％,24.30％ respectively for the patients with vaginitis.However,it was 8.80％,67.60％,8.45％ respectively for the gynecology health examination population.There is a significant difference of the total infection rate between the two population(P＜0.05).It showed no significant difference with the distribution of the two types of simple infection in the two groups ( P＞0.05).But the rate of mixing infection is much higher in the patients with vaginitis ( P＜0.05 ).Conclusion The pathology of Mycoplasma urealyticum may be related to the mixed infection of different biotypes.

OBJECTIVETo explore the correlation of the gene polymorphisms of Toll-like receptor 2 ( TLR2) and TLR4 with the susceptibility and recurrence of condyloma acuminatum (CA).

METHODSUsing Snapshot, we detected the gene polymorphisms of TLR2 597(T/C), 1350(T/C), 15607(A/G), and 2258(G/A) and TLR4 896(A/G) and 1196(C/T) in the peripheral blood of 140 CA patients and 105 HPV-negative controls. We made comparisons between the CA patients and controls as well as between the cases of recurrent CA and those of non-recurrence at 6 months after treatment.

RESULTSThere were 72, 48, and 20 cases of genotype TT, TC, and CC of TLR2 597 (T/C), respectively, in the CA patients, as compared with 71, 31, and 3 cases in the controls. The gene frequency of mutant C was 31. 43% in the patients, significantly higher than 17.62% in the controls (χ2 = 12.04, P < 0.01), and it was 38.68% in the recurrent cases, remarkably higher than 27.01% in the non-recurrent cases (χ2 = 4.16, P < 0.05). There were 74, 49, and 17 cases of genotype TT, TC, and CC of TLR2 1350( T/C), respectively, in the CA patients, as compared with 73, 29, and 3 cases in the controls. The gene frequency of mutant C was 29. 64% in the patients, significantly higher than 16. 67% in the controls (χ2 =11.05, P < 0.01), and it was 36.79% in the recurrent cases, markedly higher than 25. 29% in the non-recurrent cases (χ2 = 4.18, P < 0.05). There were 44, 66, and 30 cases of genotype AA, AG, and GG of TLR2 15607(A/G), respectively, in the CA patients, as compared with 26, 58, and 21 cases in the controls. There was no significant difference in the gene frequencies of mutant G between the two groups (χ2 = 0.33, P > 0.05). No mutant genes of TLR2 2508 (G/A) or TLR4 896(A/G) and 1196(C/ T) were detected in either the CA patients or the controls. Linkage disequilibrium analysis showed a tight linkage between TLR2 597 (T/C) and 1350(T/C) (D' = 1, r2 = 0.93).

CONCLUSIONTLR2 597(T/C) is tightly linked to 1350(T/C), which is correlated with both the susceptibility and the recurrence of condyloma acuminatum.

Aged ; Case-Control Studies ; Condylomata Acuminata ; genetics ; Gene Frequency ; Genetic Linkage ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Recurrence ; Toll-Like Receptor 2 ; genetics ; Toll-Like Receptor 4 ; genetics

The aim of the present study was to detect the transmission rate of ultrasonic low intensity pulsed ultrasound (LIPUS) through polytetrafluoroethylene (PTFE) membrane (Thickness: 0.01 mm) and Bio-Gide collagen membrane, and to provide the basis for the barrier membrane selection on the study of LIPUS combined with guided tissue regeneration (GTR). The ultrasonic (LIPUS, frequency 1.5 MHz, pulse width 200 micros, repetition rate 1.0 kHz) transmission coefficient of the two kinds of barrier membrane were detected respectively through setting ten groups from 10 to 100mW/cm2 every other 10 mW/cm2. We found in the study that the ultrasonic transmission coefficient through 0.01 mm PTFE membrane was 78.1% to 92.%, and the ultrasonic transmission coefficient through Bio-Gide collagen membrane was 43.9% to 55.8%. The ultrasonic transmission coefficient through PTFE membrane was obviously higher than that through Bio-Gide collagen membrane. The transmission coefficient of the same barrier membrane of the ultrasonic ion was statistically different under different powers (P < 0.05). The results showed that the ultrasonic transmittance rates through both the 0.01 mm PTFE membrane and Bio-Gide collagen membrane were relatively high. We should select barrier membranes based on different experimental needs, and exercise ultrasonic transmission coefficient experiments to ensure effective power.
Biocompatible Materials ; Collagen ; chemistry ; Membranes, Artificial ; Permeability ; Polytetrafluoroethylene ; chemistry ; Ultrasonics

Bone Diseases, Developmental ; surgery ; Female ; Femur ; pathology ; surgery ; Fibrosis ; surgery ; Humans ; Middle Aged

Objective To review the clinical characteristics of paraganglioma of the urinary bladder with 5 cases reprt.Methods The clinical data of 5 patients (2 males and 3 females,age 23-68 years)with paraganglioma of the urinary bladder were reported.Two cases were diagnosed by ultrasonography during health examination,1 case was found by hematuria,1 had difficulty of voiding,and 1 presented with palpitation,chest discomfort while urination.Two cases were clinical diagnosed as bladder paraganglioma,1case urachal carcinoma,and 2 cases bladder tumor.Cystoscopy showed a protruding tumor within the bladder or bladder had compressed changes.One case of tumor located in the triangle,1 in the posterior wall,1in the top,2 in the anterior wall.Three cases of biopsy were negative.Three cases of preoperative endocrine examinations showed norepinephrine significantly higher.Results Laparoscopic partial cystectomy was performed in 3 cases,open partial cystectomy in 1 case,and transurethral resection of bladder tumor (TURBT) in 1 case.One case had the complication of stress cardiomyopathy during TURBT,3 cases found intraoperative hypertensive crisis.The bleeding volume was 20-800 ml (average 126 ml),and I case received blood transfusion.During the follow-up period for 3-48 months,the blood pressure was normal,and no recurrence was found.Conclusions Bladder paraganglioma is uncommon and easily misdiagnosed.For the patients with bladder tumor,accompanied by changes in blood pressure,palpitations during urination should be highly suspicious of bladder paraganglioma.Partial cystectomy is the main treatment method.

Objective To study the proteins related to aging in aortic of old rats for laying the foundation of further study of aging mechanism.Methods The rat model of aging was built,and all model rats were divided into 4 groups:the adult group(9 weeks),the old group(12 months)of WistarKyoto (WKY) rats,and the age-matched spontaneously hypertensive rats (SHR).Blood pressure of 4 groups was observed.Morphological change of aorta was observed by HE staining.Differential proteins were identified by isobaric tags for relative and absolute quantitation,(iTRAQ)-coupled liquid chromatography and tandem mass spectrometry technology.Part of differential proteins was subsequently detected by real time PCR and Western blot.Results The mean SBP of the old group SHR was higher than WKY of 97.1％ (t=39.00,P＜0.05),and the adult group of SHR was also higher than WKY of 5.4％(t=3.64,P＜0.05).Compared with the adult group,aging change in the aortic morphology of old SHR and WKY were shown in HE staining,and the change in SHR rats was more marked.7 proteins related to aging were identified by Mass spectrum analysis,and they were Profilin-1,Prelamin A,HSP70,creatine kinase-M,Fibulin-5,eIF5A and Prohibitin.Part of differential proteins was subsequently confirmed by real time PCR and Western blot.Prelamin-A was up-regulated in the old group of WKY and SHR (0.15±0.01 vs.0.45±0.04,0.34±0.02 vs.0.78±0.06) (t=12.67,12.06,all P＜0.01),Prohibitin was down-regulated in the old group of WKY and SHR(1.34±0.05 vs.1.01± 0.06,1.24±0.05 vs.0.88±0.08) (t=7.41,7.09,all P＜0.01).Profilin-1 was up-regulated in the old group of WKY and SHR (9.12±0.4 vs.20.76±0.8,16.84±0.5 vs.55.16±0.9) (t=22.55,64.46,both P＜0.01),and Profilin-1 expression in the old group of SHR was higher thanWKY (55.16±0.90 vs.20.76±0.8,t=49.49,P＜0.01).Conclusions Differential proteins of the old rat aorta are identified through the comparative proteomics method.These differential proteins will provide new targets for the prevention and control of vascular aging.

Objective To investigate effective treatment modalities and the related factors influencing prognosis of patients with gallbladder cancer.Methods The clinical data of 76 gallbladder carcinoma patients admitted to the Department of General Surgery,PLA Nanjing General Hospital from January 2005 to October 2015 were analyzed retrospectively.Follow-up was carried out via telephone or outpatient service until January 2016.Cox regression and Kaplan-Meier models were performed for survival analysis.Results 69 patients were treated with surgery and/or postoperative adjuvant chemotherapy.The remaining 7 patients with liver or distant metastases who did not undergo surgery received chemotherapy.24 patients died from cancer relapse,37 patients died from disease progression after giving up treatment,and 7 patients were lost to follow-up.The remaining 8 patients were still alive at the time of follow-up.The depth of cancer invasion (HR =2.736),the type surgical procedure (HR =2.207),and adjuvant chemotherapy (HR =0.603) were significant impact factors of survival for GBC patients.Adjuvant chemotherapy was a protective factor.The average survival in the chemotherapy-naive group was (10.6 ± 1.9) months,the single chemotherapy group (18.5 ± 2.8) months,and the combined chemotherapy group (26.9 ± 6.4) months.There were no significant differences among these groups.Conclusions The depth of cancer invasion,types of surgical procedure particularly radical cholecystectomy,and adjuvant chemotherapy were significant factors of survival in patients with GBC.Radical cholecystectomy combined with arterial and intravenous chemotherapy using gemcitabine and oxaliplatin showed benefits in survival in GBC patients.

BACKGROUND: Induced pluripotent stem cells (iPSCs) are a special type of cells with self-renewal and multi-differentiation potential, which can differentiate into intestinal organoids under certain conditions. OBJECTIVE: To explore whether iPSCs can differentiate into intestinal organoids under specific conditions in vitro.METHODS: iPSCs from B6J mice were recovered and cultured for 3 days until clone units covered about 80％ of the culture dish, and then the cells were cultured in the medium containing Activin A for 3 days until the deterministic endoderm formed. Further, the culture medium was replaced by the medium with fibroblast growth factor 4 and Wnt3A for 4 days to differentiate into the spheroids with CDX2+. After that, spheroids were collected and mixed with Matrigel,and then the mixture was dropped into the 4-well plate and cultured with Rspondin1, Noggin, epidermal growth factor, B27 and other growth factors to differentiate into intestinal organoids. Cell morphology was observed, FoxA2 and Sox17 expresson in the deterministic endoderm was detected, and CDX2, Sox9, CGA, MMP7 were measured.RESULTS AND CONCLUSION: iPSCs were cultured with Activin A for 3 days with higher cell fusion, initial differentiation and FoxA2/Sox17 expression (P < 0.05) than those of non-induced iPSCs. Spheroids began to appear at the 3rd day after culture with fibroblast growth factor 4 and WNT3A, and formed a lot at the 4th day. And CDX2 expression in spheroids was significantly increased compared with that in the deterministic endoderm (P < 0.05). Organoids gradually formed after 3 days culture, which contained all cell types of intestinal organoids, and expressions of specific markers, Sox9, CGA, MMP7, were significantly higher than those in spheroids (P < 0.05). To conclude, iPSCs can be induced to differentiate into intestinal organoids in three-dimensional niche in vitro.

Objective To explore the relationship between lumbar disc degeneration (LDD) of lumbar spinal stenosis(LSS)and the dural sac cross-sectional area(DSCA)by MRI measurement. Methods 91 patients with central degenerative LSS were randomly selected and 91 age-and sex-matched people without LSS were select-ed as a control group. LDD was classified into five grades by MRI detection according to the method proposed by Pfirrmann and DSCA were measured. Results LDD was not associated with age in LSS. The proportion of severe degenerated disc in lower lumbar levels were higher than that of L2/3 in the two groups;DSCA in severe degenerat-ed disc group was significantly smaller than that in light degenerated group only in L2/3 and L3/4 in LSS. There were no statistical differences in every lumbar level in the control group. Conclusions LDD in L4/5 and L5/S1 of LSS is more severe than that of the normal people. DSCA and LDD are positively correlated in L2/3 and L3/4,but not in L4/5 and L5/S1 for LSS.

Renal transplant (RT) recipients have a high risk of developing cardiovascular diseases.However,the effects of renal transplantation on the development of arteriosclerosis have been controversial.The carotid intima-media thickness (CIMT) and diameter (CD) are important indicators of vascular remodeling and arteriosclerosis.In this study,31 patients with hemodialysis (HD),31 RT recipients and 84 age-and gender-matched control subjects were enrolled.Their CIMT and CD were measured by ultrasonic radiofrequency tracking,and the linear regression models and Z test were used to identify the progression of arteriosclerosis and the risk factors.Compared with HD group,RT group had significantly lower CIMT and CD.CIMT was found to be associated with age,body weight,resistance index and diastolic velocity,while CD was associated significantly with age,body weight,pulsatility index,end diastolic velocity and diastolic blood pressure (DBP),respectively.The correlation curves between CIMT and age showed the slopes of curves were decreased successively in control,RT and HD groups,and the curves between CD and age showed the slopes were decreased in order of RT ＞ control ＞ HD groups.It was concluded that CIMT and CD were significantly correlated with age in RT and moderately with age in HD patients.RT could reduce the progress of arteriosclerosis in patients with end-stage renal disease.