Objective To evaluate the possible association among serum folate,polymorphisms related reduced folate carrier gene (RFC-1) AS0G,methionine synthase reductase gene (MTRR) A66G,and cervical cancer,and to provide clues for the etiology of cervical cancer.Methods Based on a hospital-based case-control study,107 cases diagnosed as cervical cancer pathematologically and 107 controls with hysteromyoma,were selected by frequency,matched with age and habitation.Serum folate concentration was detected by RIA and polymorphism RFC-1 A80G and MTRR A66G was examed by RFLP-PCR.Results Serum folate concentration in patient group [(1.86±0.60) ng/rml] was significantly lower than that in control group [(2.30 ± 1.14) ng/ml],and risk of cervical cancer increased with the decreased serum folate levels (x2trend =12.57,P =0.001).Risks to catch cervical cancer for women with RFC-1 80 GG were 2.42 times (95 ％ CI 1.01-5.81) as much as for those with RFC-1 80 AA,and 1.65 times (95 ％ CI 0.77-3.53) for those with RFC-1 80 AA and RFC-1 80 AG,risks to catch cervical cancer for women with MTRR 66 GG were 1.35 times (95 ％ CI 0.40-4.56) as much as for those with MTRR 66 AA and 1.26 times (95 ％ CI 0.38-4.16) for those with RFC-1 80 AA and RFC-1 80 AG.Conclusion Serum folate deficiency to a certain degree can increase the risk of cervical cancer.RFC-1 A80G mutation may be a risk factor for cervical cancer and homozygous (GG) gene may increase the susceptibility of cervical cancer.MTRR A66G gene mutation may have nothing to do with cervical cancer.

Objective To summarize the clinical characteristics , diagnosis and surgical method of intra-ductal papilloma(IP)of breast without nipple discharge .Methods The clinical data of 84 IP patients(130 le-sions)without nipple discharge admitted from Feb .2011 to Oct.2013 were analyzed retrospectively .Results The age of the 84 patients were mainly ranging from 30 to 50 years old.113(86.92%)lesions were≤10 mm in size, 84(64.42%)lesions had a distance≤20 mm to nipple, 57 accompanied by adenosis , 43 accompanied by fibro-cystic adenosis , 48 accompanied by fibroadenoma , 14 with ductal hyperplasia , and 2 with atypical ductal hyper-plasia.After a follow-up of 3 to 36 months, 5 cases had recurrence , including 4 cases of IP and 1 case of ductal carcinoma in situ.Conclusions IP without nipple discharge has no typical clinical symptoms .Ultrasound exam-ination may have positive findings , but not typical .Preoperative diagnosis is difficult and surgical biopsy is rec-ommended.Multiple and atypical ductal hyperplasia has possibility of recurrence , so follow-up is necessary.

Objective:To investigate the association of plasma epoxyeicosatrienoic acids (EETs) with early neurological deterioration (END), and whether EETs are mediated by EPHX2 gene variants in patients with minor ischemic stroke (MIS).Methods:This is a prospective, multi-center observational study in patients with acute MIS in the Chinese population. Acute MIS patients with the first onset and onset within 24 hours who were admitted to Deyang People′s Hospital, the Second Affiliated Hospital of Wenzhou Medical University and the Third Affiliated Hospital of Wenzhou Medical University from March 2013 to June 2015 were recruited. Plasma EETs levels were measured on admission. Single nucleotide polymorphisms of EPHX2 gene rs751141 were genotyped using mass spectrometry. The primary outcome was END within 10 days after admission. END was defined as an increase in National Institutes of Health Stroke Scale score of 2 or more points.Results:A total of 322 patients were enrolled, of which 85 (26.4%) patients experienced END. EETs levels were significantly lower in patients with END [(60.3±7.3) nmol/L] compared to patients without END [(68.4±8.1) nmol/L , t=8.464, P<0.001]. Frequency of EPHX2 gene rs751141 GG was higher in patients with END [66/85(77.6%)] than in patients without END [123/237(51.9%),χ2=17.130, P<0.001], and patients with EPHX2 gene rs751141 GG genotype showed lower EETs levels [GG: (59.6±7.8) nmol/L, AG:(67.9±8.2) nmol/L, AA:(68.8±3.2) nmol/L, F=9.285, P<0.001]. Low level (≤64.3 nmol/L) of EETs was an independent predictor of END (31.5-51.3 nmol/L group: OR=2.96,95% CI 1.18-8.77, P=0.02; 51.4-64.3 nmol/L group: OR=2.46,95% CI 1.06-6.89, P=0.03) in multivariate analyses. END was associated with a higher risk of poor outcome (modified Rankin Scale scores 3-6) at 3 months ( OR=1.82,95% CI 1.46-2.35, P=0.02). Conclusion:END is fairly common and associated with poor outcomes in acute MIS. EPHX2 gene variants may mediate EETs levels, and low levels of EETs are related to END in acute MIS.

Objective To explore the effects of aberrant expression of DNA methyltransferase 1 (DNMT1) in cervical cancerization tissue and cervical cancer cells.Methods Cervical tissues were collected from 80 cases with a diagnosis of invasive cervix squamous cell carcinoma (SCC),53 cases with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ),52 cases with low-grade cervical intraepithelial neoplasia (CIN Ⅰ)and 53 cases with normal cervix (NC).Meanwhile,Caski (HPV16-positive) and C33A (HPV-negative) cells selected from cervical cancer cell lines were cultured routinely in vitro.The expression of DNMT1 protein and mRNA were examined by Western blot analysis and real-time PCR (qRT-PCR) in the tissues and cells,respectively.Results The levels of DNMT1 protein were 1.33,1.84 and 2.28,and the Ct-ratios (DNMT1/β-actin) of DNMT1 mRNA were 1.27,1.27 and 1.26 in CIN Ⅰ,CIN Ⅱ/Ⅲand SCC group,respectively.Comparing with NC group,the expression of DNMT1 protein or mRNA was elevated in deficient cervical groups,with statistical significance (F =110.57,P ＜ 0.001,F =2.68,P =0.048).The expression levels of DNMT1 protein were increased steadily according to severity of the cervix lesions (x2tend =50.80,P ＜ 0.001),however,the expression of DNMT1 mRNA was not observed the same tendency (x2tend =3.63,P ＞ 0.05).The results from experiment in vitro showed that the levels of DNMT1 protein or mRNA were both higher in Caski cell than in C33A cell,especially for DNMT1 mRNA with significantly difference (t =7.134,P =0.002).Conclusion Aberrant expression of DNMT1 protein or mRNA could link with the risk of cervical cancerization by both transcriptional and posttranscriptional mechanisms.There would be a synergistic effect between overexpression of DNMT1 and HPV16 infection in the progression of cervix carcinogenesis.

Objective To analyse factors associated with onychomadesis in children,and to evaluate the relationship between onychomadesis and hand-foot-mouth disease (HMFD).Methods A retrospective study was performed.Clinical data were collected from 120 children with onychomadesis (average age,3.19 ± 2.06 years) and 120 sex-and age-matched patients with hemangioma.Viral detection was realized by real-time reverse transcriptionPCR in throat swab specimens.Data were analyzed by chi-square test using the SPSS 16.0 software.Results Of the 120 patients with onychomadesis,85 (70.83％) had a history of HMFD,which was 4 to 8 weeks prior to the occurrence of onychomadesis.Onychomadesis was strongly correlated with HMFD (OR,7.621,95％ CI,4.292-13.531),but uncorrelated with the occurrence and peak of fever,abnormality of serum trace elements,existence of latent diseases,or the use of antipyretic drugs or antimicrobial drugs during the clinical course of HMFD (all P ＞0.05).Several viruses were detected in 39.7％ of 68 throat swab specimens from patients with HMFD accompanied by onychomadesis,which included Coxsackievirus A16 in 9 patients,enterovirus 71 in 3 patients and unidentified enteroviruses in 15 patients.Conclusion Onychomadesis appears to be a complication of HMFD.

Objective To explore the impact of folate on MeCP2 expression and cervical cancer cells growth.Methods Cervical cancer cell lines Caski (HPV16-positive) and C33A (HPV-negative) were treated with different concentrations of folate.MTT,flow cytometry,Western blott and real-time PCR were used to detect the cells’ viability,apoptosis,the expression of MeCP2 protein and mRNA expressions respectively.Results The inhibitions of both cell growth were upgraded with the folate concentration increasing.The differences were significant between the experimental groups and the control group.With increasing of folate concentration,apoptosis ratio of C33A and Caski increased gradually (C33A:r =0.965,P ＜ 0.001; Caski:r =0.973,P ＜ 0.001) and the expression of MeCP2 protein downgraded gradually,presenting significantly negative correlations between them (C33A:r =-0.952,P ＜ 0.001; Caski:r =-0.947,P ＜ 0.001).There was significantly difference for mRNA expression in different concentration groups of Caski and C33A (C33A:F =77.041,P ＜ 0.001; Caski:F =59.885,P ＜ 0.001).In the same concentration group,the expression of MeCP2 protein and mRNA were higher in Caski than that of C33A,and the difference was significant in the concentration of 500 μg/ml group.There was a negative correlation between the expression of MeCP2 protein and cells’ apoptosis ratio (C33A:r =-0.970,P ＜ 0.001; Caski:r =-0.93,P ＜ 0.001).Conclusion Folic acid can inhibit the growth of cerical cancer cells,promote apoptosis and reduce the expression of MeCP2.The aberrant high-expression of MeCP2 can inhibit apoptosis of Caski and C33A.

Objective The aim of this study was to investigate the expression of MAGEA4 and EB1 proteins in lung cancer tissues and their correlation with clinicopathological features and prognosis. Methods A total of 136 patients with lung cancer in our hospital were enrolled. The expression levels of MAGEA4 and EB1 at levels of mRNA and protein were measured by real-time fluo-rescence reverse transcription and immunohistochemistry. The correlation between MAGEA4 and EB1 expression and clinical patholog-ical features,and prognosis were analyzed by χ2 test and Cox regression analysis. Results The expression of MAGEA4 and EB1 mR-NA in lung cancer tissues was higher than those in adjacent tissues(P<0. 05). The positive rates of MAGEA4 and EB1 in lung canc-er tissues were higher than those in adjacent tissues(P<0. 05). The expression of MAGEA4 and EB1 proteins in lung cancer tissues was higher than those in adjacent tissues(P<0. 05). The positive rates of MAGEA4 and EB1 proteins were not significantly correlated with age(P>0. 05),but they were related to the maximum diameter,pathological grade,TNM stage,infiltration depth,lymphatic vas-cular infiltration,lymph node metastasis and recurrence(P<0. 05). The 3-year survival rate and total survival time of MAGEA4 and EB1 negative group were significantly higher than those of MAGEA4 and EB1 positive group(P<0. 05). Lymphatic vascular infiltra-tion,lymph node metastasis,MAGEA4 positive and EB positive were independent risk factors for prognosis of patients with lung cancer (P<0. 05). Conclusion The positive expression rates of MAGEA4 and EB1 proteins in lung cancer tissues are increased,and their high expression may be related to the occurrence and development of lung cancer. Lung cancer patients with negative expression of MAGEA4 and EB1 proteins can obtain better prognosis.

OBJECTIVETo investigate the curative effect of penile elongation with four differentoperative approaches.

METHODSThrough four different operative approaches (the coronary sulcus ringincision, Y or Z shaped incision or Z shaped incision combined with coronary sulcus ring incision), thepenile skin and fascia were degloved until the penile root. Then the superficial and deep dorsal penilesuspensory ligament were cut off. After electric coagulation of the residue ends, the two-side tissue at thefront of the pubic symphysis was sutured. Then the penile skin and fascia were repositioned and the incisionat the inner and outer plate was closed.

RESULTSThe increased penile static length was (2.9 ± 0.2) cmwith abdominal wall Y incision (12 cases); (3.1 ± 0.3) cm with transabdominal modified Z incision (260 cases); (3.9 ± 0.7) cm with coronary sulcus ring incision (363 cases); (3.4 ± 0.8) cm with combined incision (39 cases). The lengthening effect was significantly different between the coronary ring incision and abdominal wall Y/Z incision (P < 0.05). The postoperative follow-up period was 6 months to 5.5 years without serious complications. Only 3 cases of subcutaneous hematoma occurred with treatment of debridement and drainage. 4 cases with ischemic necrosis at distal penile skin, were treated with debridement, dressing and physiotherapy, leaving no scar.

CONCLUSIONSPenile lengthening surgery are safe and effective through different approaches. The coronal ring incision has the best therapeutic effect.

Bandages ; Debridement ; Drainage ; Electrocoagulation ; Fasciotomy ; Follow-Up Studies ; Humans ; Ligaments ; surgery ; Male ; Necrosis ; surgery ; Organ Size ; Penis ; anatomy & histology ; pathology ; surgery ; Postoperative Period ; Reconstructive Surgical Procedures ; methods ; Skin ; Time Factors

With the development of molecular biology, biomaterials and tissue engineering, regenerative treatment of pulpal and periradicular diseases is facing new opportunities. At present, a large number of studies on dental pulp regeneration reveal that cytokines are essential for promoting migration, proliferation and osteogenic differentiation of dental pulp stem cells. In this paper, we review several kinds of cytokines related to dental pulp regeneration, and analyze their roles and regulatory mechanisms in dental pulp regeneration.

Objective: To establish an accurate and selective UPLC-MS/MS) method for the determination of afatinib in rat plas-ma. Methods: Protein precipitating by acetonitrile was used to prepare the samples. A CORTECS BEH C18column ( 50 mm × 2. 1 mm, 1. 6 μm) was used to separate the analytes at 40℃. The mobile phase consisted of acetonitrile and water (0. 1% formic acid) with the flow rate of 0. 4 ml·min-1. The analytes were quantified by multiple reaction monitoring ( MRM) mode with positive electrospray ionization, while the target fragment ions were m/z 486. 19→112. 1 for afatinib and m/z 557. 3→112. 15 for neratinib (IS). Results: The calibration curve obtained good linearity for afatinib within the range of 1–200 ng·ml-1(r=0. 998 1), and the LLOQ in rat plasma was 1. 0 ng/ml. The intra-and inter-day precisions were both≤9. 51% . The recovery of afatinib from plasma was above 77. 1% . After intragastric administration and intravenous administration of afatinib in rats, the t1/2was 7. 19 h and 2. 69 h, Cmax was 97. 78 ng·ml-1and 123. 37 ng·ml-1,and AUC(0-∞)was 1 505. 4 ng·ml-1·h and 405. 55 ng·ml-1·h, respectively. Con-clusion: The validated method can be applied in the pharmacokinetic study of afatinib at the intragastric and intravenous dosage of 10 and 2 mg·kg-1, respectively.