AIM:To investigate the role of bcl-2 and bax expression in apoptosis of pulmonary artery smooth muscle cells(PASMC) of rats induced by Na +/H + exchanger isoform-1(NHE-1) inhibition in vitro. METHODS:Intracellular Ca 2+ concentration ([Ca 2+ ]i) was measured using Fura-2/AM. The expression of bcl-2 and bax mRNA in cells were detected by sqRT-PCR, and the expression of Bcl-2 and Bax protein were examined immunohistochemically. RESULTS:The expression of bax mRNA and Bax protein and [Ca 2+ ]i in cells transfected with NHE-1 ribozyme gene increased significantly compared with those of cells transfected with pLXSN and nontransfected control. Meanwhile, the expression of bcl-2 mRNA and Bcl-2 protein in cells transfected with NHE-1 ribozyme gene decreased significantly. CONCLUSION:Apoptosis of pulmonary artery smooth muscle cells induced by inhibiting NHE-1 may have relevance to the increase in [Ca 2+ ]i and bax expression and the decrease in bcl-2 expression.

Objective To investigate the clinical features of skin cancer.Methods Clinical data of skin cancer and precancerous skin lesions confirmed pathologically from 2005 to 2008 in Peking University First Hospital were retrospectively analyzed by using statistical methods.Results A total of 632 cases of skin cancer and precancerous skin lesions were studied.The most common skin cancer was basal cell carcinoma and squamous cell carcinoma (invasive and in situ) which accounted for 29.3%and 24.2%,respectively.The average age at onset was older than 60 years in 55.4%of the patients,between 35 and 59 years in 34.3%,younger than 35 years in 10.3%.The concordance between clinical and pathological diagnosis reached nearly 90.O%for Paget's disease,70.0% for other common skin cancer and precancerous skin lesions.Conclusions Skin cancer and precancerous skin lesions have a predilection for scalp and face.Patients aged from 35 to 59 years account for a significant proportion not only in cutaneous lymphoma but also in melanoma and epithelium-derived nonmelanoma skin cancer.

Objective To discuss the clinical effects of transverse screw fixation of double columns in crescent pelvic fractures.Methods Twenty patients with crescent pelvic fractures hospitalized from December 2012 to December 2015 were reviewed retrospectively.There were thirteen male and seven female patients with the age ranging from 22 to 72 years (mean,39.5 years).Causes of injury were traffic accidents in fifteen patients,falling from high places in three and hitting by heavy objects in two.Time interval between injury and operation was 3-14 d.According to the Orthopedic Trauma Association (OTA) classification,all were classified as 61-B2 type.Reconstruct plate was used to stabilize the supperior pubic ramus fractures,and transverse double-column fixation with anterograde or retrograde screws was used for the posterior iliac crescent fractures.Duration of inserting screws,operation time,intraoperative blood loss and frequency of C-arm X-ray were recorded.Radiological and clinical outcomes were evaluated after operation.Results Duration of inserting screws ranged from 3 to 16 min (mean,5 min).Operation time ranged from 50 to 130 min (mean,80 min) and the intraoperative blood loss ranged from 200 to 550 ml (mean,280 ml).Frequency of C-arm X-ray in the surgery ranged from 1 to 5 times (mean,2 times).All screws were in the expected location,without any piercing out of the iliac bone dependent on the confirmation of X-ray and CT-scan after operation.According to the Matta and Tornetta radiological evaluation,the reduction was rated excellent in sixteen patients and good in four,with the excellent-good rate of 100％.There was no death,wound infection,sciatic nerve or superior gluteal nerve injury or deep venous thrombosis of lower extremities.At the followup,no nonunion,loss of reduction,and breakage of internal fixation occurred.Nineteen patients were followed up,and mean follow-up time was 19.6 months (range,3 to 36 months).Union was obtained in all patients in a period of 8-17 months (mean,11.2 months).According to the Majeed functional evaluation at the final follow-up,the outcome was rated excellent in seventeen patients and good in two,with the excellent-good rate of 100％.Conclusion Transverse screw fixation of double columns in crescent pelvic fracture of 61-B2 type can reduce the operation injury,decrease complications,and have good clinical results.

Objective To construct a large animal model of ischemic heart disease induced by acute myocardial infarction (AMI) . Methods Male Diannan Mini-pigs were selected (n=8), weight 20 ± 4 Kg. After general anesthesia and endotracheal intubation, blood pressure and electrocardiograph were monitored. Parasternal incision was taken and thoracotomyv was performed at 4th-to-5th intercostalgap. Pericardium was opened and left anterior descending artery was ligated. Echocardiography was performed at baseline, postoperative 28 and 60 day. Euthanasia and heart explanted was conducted at 60 day of experimental pig. After gross examination of the heart, the histological examination was used to assess myocardial infarction by using H&E staining. Results All of the pigs were presented with ventricular arrhythmias in varying degrees after ligation of the left anterior descending artery, of whom 2 died of refractory ventricular fibrillation, the mortality was 25%. Echocardiography assessment showed the loss left ventricular function, the thinning of left ventricular apical and the enlargement of left ventricule. Histological examination revealed that myocardial fibrosis and fibrous scar were formed in myocardial infarction area. Conclusion The ligation of left anterior descending artery through thoracotomy under direct vision is a reliable strategy for construction of AMI porcine model and the experimental porcine can develop ischemic cardiac dysfunction progressively.

Objective To evaluate the clinical efficacy of universal self-locking anatomical plate for acetabulum and pelvis (USAPAP) in the treatment of posterior acetabular fractures.Methods Data of 55 patients with posterior acetabular fractures who were treated with the USAPAP from January 2014 to January 2016 were retrospectively analyzed.There were 39 males and 16 females with an average age of 38.5 years old (range,17-82 years).There were 35 fractures on the left side while the other 20 fractures were on the right side,including 52 fresh cases and 3 old cases.According to Letournel-Judet classification,there were 32 cases of posterior wall,9 cases of transverse and posterior wall,5 cases of anterior column and posterior wall,2 cases of posterior column,2 cases of transverse,2 cases of two columns,1 case of anterior column and posterior hemitransverse,2 cases of Pipkin type Ⅳ.All patients were managed operatively by the USAPAP,which allows simultaneous fixation for two columns and quadrilateral surface fractures through a single Kocher-Langenback approach.The quality of reduction was assessed by Matta's score system.The mean follow-up period was 18.5 months (range,12-24 months).Average operation time was 95 min,and average blood loss was 350 ml.Average operation time of plate and screws fixation was 19 min,and average fluoroscopy times in the surgery was 2 times.The mean time of bony union was 4.3 months.According to the criteria described by Matta,the excellent and good rate of radiological score was 92.7％ (51/55),including 39 cases excellent,12 cases good and 4 cases poor.The excellent and good rate of Merle d'Aubigne-Postel score was 87.3％ (48/55),including 35 cases of excellent,13 cases of good,5 cases of fair,and 2 cases of poor.Heterotopic ossification occurred in 1 patient (Brooker type Ⅱ) at three months postoperatively.One case with Pipkin type Ⅳ fracture underwent total hip arthroplasty due to femoral head necrosis at seven months postoperatively.Complications including wound infection,sciatic nerve injury,screw invade joint,redisplacement of the fracture,avascular necrosis of bone fragment,hardware failure,nonunion or malunion was not found in any case at the latest follow-up.Conclusion The USAPAP through the single posterior Kocher-Langenback approach provides strong and stable fixation for complex acetabular fractures associated with posterior region including posterior wall,both columns and quadrilateral surface.Satisfactory clinical results can be achieved by the use of this method.

Objective To evaluate the relationship between desmosome-related proteins and epidermal tumors. Methods Using anti-desmoglein 1 and 2 monoclonal antibodies, expression of desmoglein 1 and 2 was examined by an immunohistochem ical staining method in various epidermal tumors such as squamous cell carcinoma (SCC), actinic keratosis(AK), keratoacanthoma(KA) and seborrhoeic keratosis(SK). Results Desmoglein 1 and 2 were strongly expressed in intercellular space of wh ole epidermis in normal human skin. Expression of desmoglein 1 and 2 was markedl y reduced or absent in SCC cells. Compared with normal epidermal cells, expressi on of desmoglein 1 and 2 was equal to or reduced,with complete absence of stain ing in dysplastic areas in AK cells. Extensive pericellur staining of desmoglein 1 and 2 was found in KA and SK cells, similar to that observed in normal epider mis. Conclusion Expression of desmoglein 1 and 2 is markedly reduced or absent i n human malignant epidermal cancers, reduction of these molecules may contribute to invasiveness and metastasis of epidermal carcinomas.

Objective To construct a colon cancer chemotherapy-resistant cell line COLO,and study its characteristics and its relationship with tumor stem cells.Methods We constructed two 5-fluorouraci (5-FU)-resistant colon cancer cell line COLO/5-FU-1 and COLO/5-FU-2, which were resistant to 0.1 0 μmol/ml and 0.20 μmol/ml 5-FU respectively through gradiently increased drug concentration.The cha-racteristics of 5-FU-resistant cell lines were compared with parental colon cancer cell line COLO related to proli-feration,colony forming ability,migration and invasion,sphere forming ability,expression of stemness genes and cross drug-resistance.Results In the cell viability assay,4 days after regular training,the absorbancy of colon cancer 5-FU-resistant cell lines COLO/5-FU-2,COLO/5-FU-1 and parental colon cancer cell line COLO were 0.61 ±0.1 3,0.54 ±0.07 and 0.41 ±0.09 respectively,with significant difference (F =63.43,P =0.033).With the increased concentration of 5-FU,5-FU-resistant cell lines presented increasing clonality. The cloning efficiency of COLO/5-FU-2,COLO/5-FU-1 and parental colon cancer cell line COLO were (87.6 ±1 2.7)%,(65.3 ±9.7)% and (38.5 ±7.6)% respectively,with significant difference (F =33.64, P =0.01 7).In each high power field of vision,the cell numbers of migration through the basement membrane of COLO/5-FU-2,COLO/5-FU-1 and parental colon cancer cell line COLO were 482 ±39,434 ±45 and 373 ±38 respectively;and the cell numbers of invasion through the basement membrane were 1 74 ±42,1 1 2 ± 31 and 87 ±29 respectively,with significant differences (F =1 09.61 ,P =0.009;F =67.31 ,P =0.032). Compared with parental colon cancer cell line COLO,5-FU-resistant cell lines had higher expression of stem-ness genes (F =47.31 ,P =0.042).5-FU-resistant cell lines were cross-resistant to other chemotherapeutic drugs such as mitoxantrone.For example,after incubation for 96 hours,inhibition rate of mitoxantrone to parent colon cancer cell line COLO was higher significantly than COLO/5-FU-1 and COLO/5-FU-2 (0.749 ± 0.042,0.423 ±0.024,0.342 ±0.01 8),with significant difference (F =1 2.61 ,P =0.028).The micro-sphere forming rates of COLO/5-FU-2,COLO/5-FU-1 and parental colon cancer cell line COLO were (8.90 ± 0.97)%,(6.20 ±0.75)% and (3.90 ±0.32)% respectively,with significant difference (F =1 64.32,P =0.006).Conclusion Colon cancer drug-resistant cell line COLO possess tumor stem cell-like characteristics, which are enriched in cancer stem cells.

Objective To evaluate the sensitivity and specificity of BP180NC16a-ELISA in the diagnosis of bullous pemphigoid (BP). Methods A multi-center, randomized, double-blind, parallel-controlled study was conducted. Sera were collected from 106 patients with clinically confirmed active BP and 106 control subjects including patients with non-BP bullous diseases, scleroderma, psoriasis or systemic lupus erythematosus,late pregnant women and healthy blood donors. BP180NC16a-ELISA was performed on these sera. The IgG antibody levels measured by ELISA kit were compared with those measured by indirect immunofluorescence (IIF) test. Results Of the 106 BP sera, 81 were positive for BP180NC16a-ELISA with a sensitivity of 76.4%,83 for ⅡF test with a sensitivity of 78.3%. Among the 106 control serum samples, 95 were negative for BP180NC16A-ELISA with a specificity of 89.6%, and 102 for ⅡF test with a specificity of 96.2%. There was no significant difference between the two tests in dignostic sensitivity and specificity for BP (both P ＞ 0.05).Conclusion BP180NC16A-ELISA may serve as an adjuvant tool for the diagnosis of BP.

Objective:To investigate the short term outcomes and postoperative respiratory complications of patients with chronic thromboembolic pulmonary hypertension(CTEPH) treated by pulmonary endarterectomy(PEA).Methods:45 consecutive CTEPH patients underwent PEA between December 2017 and January 2020 in our institution were enrolled, including 25 females and 20 males. The mean age of operation was 51.2(25-70) years old. 24(53.5%) patients were in New York Heart Association(NYHA) functional class Ⅲ-Ⅳ. The mean PVR before operation was 923(461-2 711) dyn·s·cm -5. All patients’ data were entered in a prospective database, divieded into patients with respiratory complications group(WRC)and without respiratory complications group(WORC). To assess risk factors for postoperative respiratory complications and its effect on short term outcomes. Results:There was a significant reduction in mPAP(from 37 mmHg to 20 mmHg) and PVR(from 923 dyn·s·cm -5 to 293 dyn·s·cm -5) in the entire group. The in-hospital mortality rate was 4.4%(2 cases), died due to postoperative cardiogenic circulatory failure, even with VA-ECMO treatment and mediastinal infection, respectively. Postoperative respiratory complications occurred in 32 patients(71.1%). The most common complications were reperfusion pulmonary edema 44.4%(20 cases) and residual pulmonary hypertension 11.1%(5 cases). The WRC group showed a tendency to have longer periods of mechanical ventilation, longer ICU stays and more ICU costs. Independent predictors of postoperative respiratory complications were time from symptom onset to PEA>36 months( OR=12.2, 95% CI: 2.1-70.7, P=0.005)and six-minute walking distance<300 m( OR=12.6, 95% CI: 1.1-138.0, P=0.0038). Conclusion:Pulmonary endarterectomy is an effective and safe treatment for CTEPH. Postoperative respiratory complications were mainly determined by symptom onset time and pre-operative status. Patients with CTEPH should consider PEA surgery early.

Objective:To investigate the effect of high-frequency repetitive transcranial magnetic stimulation(rTMS)on cue attention in male patients with alcohol use disorder (AUD) after the acute withdrawal.Methods:A total of 90 male patients AUD who were hospitalized in the Second Affiliated Hospital of Kunming Medical University and Psychiatric Hospital of Yunnan province from May 2020 to December 2020 were enrolled, then they were divided into study group and control group using random number table.Because 18 cases fell out during the study, 36 cases were included in each of the two groups.After the alcohol withdrawal syndrome eliminated, the study group received high-frequency rTMS at 10 Hz for 14 consecutive days, and the control group was administrated by sham rTMS.At baseline and after true or sham rTMS, the cognitive psychology experiment Oddball paradigm was completed, and the behavioral data of the subjects were collected.Paired-sample t-test was used to compare the changes of the two groups before and after treatment.Data analysis were conducted using SPSS 21.0 software. Results:There was no statistical difference between the study group and the control group in terms of drinking level, cognition level and demographic data(all P>0.05). In the Oddball paradigm, compared with the control group((526.72±75.30)ms, (0.98±0.02))the reaction time((497.93±64.51)ms, t=3.145, P=0.008) and accuracy rate((0.99±0.01), t=-2.803, P=0.016) in alcohol-related cues were significantly improved in the study group after rTMS intervention, but in the control group, there were no statistical differences(both P>0.05), whether the cue was alcohol related or not. Conclusion:The results suggest that the rTMS can enhance the attention bias of alcohol-related cues and change the impulse process partly.