A novel PCR-based mutagenesis method was reported, in which there is no need to purify megaprimers or design a special flanking primer. This method used one mutagenic primer and two sequencing primers (T_m≤58℃) as flanking primers. After first round PCR, 12.5 μl first PCR production was directly added into 50 μl second PCR system as template and megaprimer, and 10 rounds of asymmetrical PCR at high temperature of annealing (68 ℃ ) was to add in initiation of second PCR. This additional step greatly has increased the efficiency of mutagenesis via 600 bp or 800 bp long megaprimer. The results demonstrated that this method can achieve high fidelity, 97%～ 98% efficiency, high yield.

Objective To assess dermatoscopic characteristics of lichen sclerosus(LS). Methods Dermatoscopy was performed to observe 27 genital or extragenital porcelain-white skin lesions in 15 outpatients with confirmed LS. Results As dermatoscopy showed, of the 27 skin lesions, 24 showed white-yellowish structureless areas, 25 linear vessels, 27 white patches, 17 keratotic plugs. Characteristic cloverleaf-like structure was observed in 7 skin lesions in 4 patients, which was consistent with LS. Conclusion Patients with LS show atypical clinical manifestations, but specific dermatoscopic patterns.

Objective To study the consistency and complementarity of endoscopic ultrasonography (EUS),white light endoscopy (WLE) and magnifying endoscopy (ME) in diagnosis of ulcerative colitis (UC).Methods We collected 125 cases of UC patients diagnosed by WLE and EUS (including 51 cases of WLE + ME + EUS).According to UC mucosal morphology under WLE and crypt openings under ME,we divided all the cases into several groups and analyzed intestinal wall thickness (TWT) under EUS in each group.Results According to the results of UC inflammation degree under WLE,all patients were divided into four groups: 16 severe cases,46 moderate cases,44 mild,and 19 remission stage.TWT results were (5.903 ± 1.551 ) mm,(4.673 ± 1.235 ) mm,(3.756 ± 1.322 )mm and ( 3.464 ± 0.970) mm,respectively.Differences were significant between any two groups ( P ＜ 0.05 ),except for that between mild and remission groups.According to the results of UC inflammation degree under ME,all patients were divided into six groups: 9 cases of villous-like structure,9 cases of typical coral reef-like structure,8 severe coral reef-like structure,13 regular crypt opening,6 epithelial minimal defect and 6 small yellow spot (SYS).TWT results were (5.701 ±0.941 )mm,(5.518 ±0.581 )mm,(5.181 ±0.751 )mm,(3.763 ±0.659) mm,(3.587 ±0.461 )mm and (2.505 ± 0.330 )mm,respectively.Differences were significant between any two groups ( P ＜ 0.05 ) except for those between epithelial minimal defect and regular crypt opening,typical coral reeflike structure,villous-like and severe coral reef-like structure.EUS results showed SYS (6/6) and regular crypt opening ( 10/13 ) were mostly located in mucosa,while lesions of severe coral reef-like structure (8/8) invaded the muscularis propria.Conclusion EUS shows high consistency with WLE and ME in diagnosis of UC inflammation degree and invasive depth.It could assist and even substitute ME for evaluation.

Objective To investigate the effects of hyperbaric oxygen preconditioning (HBOP) on brain edema, inflammatory reaction and neuronal cell apoptosis induced by experimental hemorrhage in rats. Method Eighteen male Spraque-Dawley rats, weighing 300 - 350 g,received five successive sessions of HBOP with 3 atmosphere absolute pressure and 100% O2 one hour daily for five successive days, and other eighteen rats received five successive sessions of pretreatment with one atmosphere absolute pressure, air, one hour daily for five successive days. Twenty-four hours after the final pre-conditioning, rats received an infusion of 100 μL autologous blood into the basal ganglion. Seventy-two hours later, rats were sacrificed for brain edema measurements in 12 rats of each group. The histopathological changes around the hematoma were observed microscopically, and the neuronal cell apoptosis was detected by using the terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) in six rats of each group. Data of brain water content were analyzed by using Stata 7.0 software and statistical analysis was carried out by two-tailed Student t -test. Results Compared with the control group, HBOP significantly attenuated brain edema 72 hours after intra-cerebral hemorrhage in experimental rats (81. 6± 0. 7% vs. 82. 8± 0.9%, P < 0.01). Inflammatory cell infiltration and neuronal cell apoptosis were also significantly decreased in the HBOP group. Conclusions HBOP protects the rats against brain edema formation, and quells inflammatory reaction and neuronal cell apoptosis following intra-cerebral hemorrhage in experimental rats.

Objective To discuss the influence on peripheral circulation and oxygenation of different colloid osmotic pressure (COP) in pediatric cardiac surgery under cardiopulmonary bypass (CPB).Methods Sixty cases of non-cyanotic congenital heart disease patients under 10 kg were randomly selected and divided into 3 groups(n =20) according to the different COP level.COP values was adjusted by the ultrafiltration technique and colloid addition.The perioperative(T1-T6) arterial lactate level,different value between skin and rectal temperature,peripheral oxygen saturation (SpO2) and oxygenation index (OI) were observed in order to determine the different effect on peripheral circulation and oxygenation.Meanwhile,mechanical ventilation time and ICU time were recorded.Results The variation tendency of arterial lactate level was similar in each group,the value in the COP ＞ 18 mmHg (1 mmHg =0.133 kPa) group(group C) was significantly higher than COP 10-15 mmHg group (group A) and COP 16-18 mmHg group (group B) in T3 and T4,after CPB weaned,the values of Group A (1.25 ± 0.42) and Group C (1.33 ± 0.51) were higher than Group B (0.71 ± 0.29) at T6 point (P ＜ 0.05);the variation tendency of SpO2 was similar in each group too,the value of group C was significantly lower than group A and B at T5 point,the values of group A and C were significantly lower than group B at T6 point,P ＜ 0.05;the different value between skin and rectal temperature in group A was significantly higher than group B and C from T1 to T2 point(P ＜0.05),but not in T3 to T6 point;The minimal OI values of all the groups were appeared in T4 point,group B value was significantly higher than A and C in all time point,group C value was the lowest(P ＜0.05);the mechanical ventilation time in group B(2.13 ± 1.36) days and group C (2.93 ± 1.69) days were significantly lower than group A (3.83 ± 1.47) days,P ＜ 0.05.ICU time of group B (3.9 ± 1.1) days was significantly lower than group A (5.7 ± 2.5) days and C (6.0 ± 1.5) days.Conclusion During the pediatric CPB,the improper COP level will lead to bad oxygenation and poor peripheral circulation,got different prognosis ultimately.A reasonable COP level(16-18 mmHg) will do benefits to all the pediatric patients.

Objective To study the clinical features,diagnosis and management of small cell carcinoma of the bladder (SCCB).Method The clinical data of 18 cases of patients with small cell carcinoma of the bladder were analyzed retrospectively and the literature were reviewed.There were 16 males and 2 females,ages 54 to 81 years (median age,61 years).Clinical manifestations included gross hematuria in 11 cases,urgency in 2 cases,dysuria in 2 cases and postoperative review after TURBT of bladder urothelial carcinoma in 3 cases.The median tumor size was 3.35cm (ranged,1.0 to 6.0 cm).2 cases underwent TURBT and intravesical chemotherapy regularly were followed after surgery.3 cases underwent partial cystectomy,intravenous chemotherapy combined with radiotherapy was followed in one case,the other 2 cases refused the following therapy.13 cases underwent radical cystectomy,intravenous chemotherapy was followed in 2 cases,pelvic radiotherapy was followed in 2 csaes and intravenous chemotherapy combined with radiotherapy was followed in 2 cases,the other 7 cases refused the following therapy.Results 11 cases were pure SCCB,7 cases were mixed SCCB,all with urothelial carcinoma.T1N0M0 in 3 cases,T2N0.1M0 in 4 cases,and T3N0-2M0 in 11 cases.The duration of follow-up was from 5 to 35 months after surgery.9 cases died of tumor metastasis,9 cases are still alive,except 1 case with lymph node metastasis,the other 7 cases are free of tumor recurrence or metastasis.Conclusions SCCB is rare,with high malignant degree and poor prognosis.The prognosis of the pure SCCB may be worse than the mixed SCCB.The diagnosis depends on pathology examination.Radical cystectomy is the main treatment method,the strategy of bladder-preserving may be an attempt for proper SCCB patients.Adjuvant therapy plus surgery may be better.

Objective ERP research has always been the hot spot of the n~ve electrophysiology research.A gTowing body of research suggests that ERP can effectively reflect the cognitive function in patients with depression,evaluate the risk of suicide.This paper reviews the ERP related components with depression and related research.On the basis of the application of ERP in the depression research discusses limitations and prospects on the later development.Methods The articles were retrievaled from databases,such as P UBMED,Foreign medical information resources retrieval platform,CNKI,Wanfang database,ect,from 2013-06 to 2015-01.Index strategy TI:TI:[depression OR affective disorder] AND [suicide OR suicidal] All field:event related potential.Reading and narrowing the scope of the articles,we should understand the experimental design,such as Sample company have suicidal ideation and (or) behavior of patients with depression.Potential induced way,ERP related component detection method,etc.The articles included 43 in English and 15 in Chinese.Read the full text and retain nearly five years about depression and related literature.Results Eventually,the article includes 34 articles and 3 of in Chinese.Conclusion Associated with suicidal ideation and (or) behavior of the existing cognitive dysfunction in patients with depression,ERP related components such as:abnormal P300 and CNV,MMN.N400 have not found abnormal.This article only from the Angle of depression and related research are reviewed in this paper the characteristics of ERP,ERP of other mental illnesses among related research,and to be perfect in the future.

Objective Recent years,epigenetics research mainly focused on DNA methylation.A growing body of research indicated that DNA methylation may play vital role in etiology of affective disorder.With brief introduction of DNA methylation molecular mechanisms and relevance of DNA methylation to affective disorder,this review focuses on some supporting evidence which are recently emerged in major depressive disorder (MDD) and bipolar disorder related gene DNA methylation.Methods We retrievaled related articles from databases,such as PUBMED,Foreign medical information resources retrieval platform,CNKI,Wanfang database,ect,from 2013-06 to 2015-01.Index strategy TI:[depression OR bipolar disorder OR affective disorder] AND All field:DNA methylation.Reading and narrowing the scope of the articles,the experimental design was understanded,such as DNA from either blood cell or brain parenchyma of sample,methods of methylation detection,gene detected and methylation position,etc.The articles included 168 in English and 32 in Chinese.The whole and retained the articles were red about MDD and bipolar disorder related gene DNA methylation in recent five years.Results Eventually,the article includes 29 articles and two of in Chinese.Most of them are receptor gene epigenetics research associated with disease.Conclusion MDD and bipolar disorder are associated with the higher of DNA methylation levels in multiple gene promoter region,such as GR,BDNF,5-HTR,SLC6A4 and the lower of COMT.This article only from the perspective of affective disorder reviewed the relationship between DNA methylation and diseases.None of other diseases epigenetics research,there is to be perfected in future.

Objective To investigate the role of Heme oxygenase-1 in the effect of hyperbaric oxygen preconditioning (HBOP) against the brain edema formation after experimental intracerebral hemorrhage in rats.Methods The study was carried out by animal experiment in two steps by using 54 Spradgue-Dawley rats weighting from 300-350 g.In the first step,rats were treated with HBOP (HBOP group,n =3) or with sham pre-conditioning (Sham pre-conditioning group,n =3).All the rats were sacrificed 24 h after the preconditioning,and basal ganglion of brain tissue was taken for detect HO-1 level by using western blot analysis.In the second step,rats were divided into 4 groups (n =12 in each group):HBOP +ZnPP group,in which rats had a micro-pump intra-peritoneally implanted containing a specific HO-1 inhibitor ZnPPⅨ (Zinc protoporphyrin IX,0.01 mg/kg),Sham pre-conditioning + Znpp group,HBOP + DMSO group,in which rats with a intra-peritoneal micro-pump containing 2 mL Dimethyl sulfoxide (DMSO,a solvent vehicle) and Sham pre-conditioning + DMSO group before HBOP.At 24 hours after the pre-conditioning,rats received an infusion of 100 μL autologous blood into the caudate nucleus to form a simulated intracerebrum hemorrhage (ICH),and were sacrificed 72 h later for brain water content measurements.All data were analyzed by using Stata 7.0 software and statistical analyses were carried out by two-tailed Student t test.Results Compared with the Sham pre-conditioning group,the HBOP group had significant higher level of HO-1.Compared with the Sham pre-conditioning + DMSO group,the HBOP + DMSO group had a significant lower level of water content in the ipsilateral basal ganglion [(81.4 ± 0.9) ％ vs.(82.6 ± 0.8) ％ (P ＜ 0.05)],however,peritoneal infusion of ZnPP Ⅸ before HBOP abolished HBOP-induced protection against brain edema formation after experimental ICH [(82.8 ± 0.9) ％ vs.(82.6 ± 0.7) ％ (P ＞ 0.05)].Conclusions Hyperbaric oxygen preconditioning attenuate brain edema formation after experimental ICH in rats,and this protection is attributed to the activation of HO-1.

Aim To investigate the protective effect of rosiglitazone on cerebral ischemia/reperfusion injury inrats. Methods Transient focal cerebral ischemia injury model in rats was induced by occlusion of the right middle cerebral artery for 2 h, followed by 24 h reperfusion. The infarct volume and neurological deficit were determined by the method of TTC staining and the Longa′s score, and used to evaluate the effect of rosiglitazone on cerebral injury. The levels of malondialdehyde (MDA), nitric oxide (NO), activities of superoxide dismutase (SOD), myeloperoxidase (MPO) and nitric oxide synthase (NOS) in brain were measured by spectrophotometer. Immunohistochemistry was employed to assess the expression of intracellular adhesion molucule-1 (ICAM-1). The histopathological change was observed after HE staining. Results Pretreatment with rosiglitazone markedly reduced brain infarct volume and neurological deficit induced by transient ischemia, inhibited MPO activity, as well as expression of ICAM-1; it also decreased NO, MDA levels and NOS activity, increased SOD activity, and improved histopathological injury. Conclusion Rosiglitazone has a protective effect on cerebral ischemia/reperfusion injury through inhibiting inflammatory process and lipid peroxidation.