Objective To study the in vitro culture condition for melanoblasts from human foreskin tissue. Methods The skin tissue taken from foreskin of children was treated with 0.5% dispase Ⅱ to separate epidermis from dermis, then with trypsin to obtain single cell suspension, which was cultured in modified medium for melanoblasts, i.e., MCDB254 medium supplied with several cell growth factors. Finally, melanoblasts were obtained based on the difference of adhesion speed. The morphology and proliferation of cultured melanoblasts were observed under a light microscope. DOPA staining, immunostaining with anti- S-100 and -tyrosinase related protein 2 (TRP2) antibodies, and transmission electron microscopy were per- formed to identify the cultured melanoblasts. Results The cultured human melanocytes displayed a match-like shape, scattered arrangement, syrmnetric double poles, slim cell body, highly refractive nuclei; meanwhile, the melanoblasts exhibited plentiful cytoplasm, large volume, bipolar or irregular shape and clonal growth. Additionally, the melanocytes were positive for TRP2, S-100 and Dopa staining, while the melanoblasts were positive only for TRP2. Electron microscopy revealed the presence of mature melanin granules (stage Ⅲ-Ⅳ ) in melanocytes but immature melanin granules (stage Ⅰ ) in melanoblasts. Conclu- sion Stable pure culture of melanoblasts has been realized with the reformed medium, which may lay a foundation for the investigation into the mechanism of epidermal pigmentation.

Aim Bupivacaine is a kind of long-acting amide local anesthetics.This paper aims to explore the effects of bupivacaine on the short-circuit currents in human alveolar epithelial monolayers and study the possible mechanisms.Methods Short-circuit currents were recorded by ussing-chamber setup.Amiloride-sensitive currents were defined as the difference be-tween the total current and the amiloride-resistant cur-rent.ERK1 /2 phosphorylation protein levels were ana-lyzed by Western blot at 0,1 5,30 and 60 min after administration of 1 00 μmol·L -1 bupivacaine.Results Bupivacaine could inhibit the short-circuit currents in H441 monolayers dose-dependently,which could be inhibited by amiloride.Western blot analysis showed that bupivacaine increased the level of ERK1 /2 phos-phorylation.Conclusion These data demonstrate that bupivacaine can reduce the alveolar ion transport by in-hibiting the amiloride-sensitive currents,possibly by the enhancement of ERK1 /2 phosphorylation. The effects of alveolar fluid clearance following application of bupivacaine should be considered clinically when the patient is complicated with lung injury.

Objective To investigate the usefulness of acoustic radiation force impulse (ARFI) elastography for noninvasive evaluation of liver fibrosis in rats.Methods A total of 70 male Wistar rats were included in the group for dimethylnitrosamine (DMN)-induced liver injury,and 10 saline-injected rats were used as normal control.Hepatic injury was induced by a single intraperitoneal injection of DMN at a dose of 50 mg/kg of body weight.Several rats in the group with DNM injected and the normal control group were randomly selected and sacrificed at each of the following post-injection time:day 5,7,10,14,21,24,and 28.And their livers were taken for pathology analysis.All the rats underwent ARFI elastography before sacrificed in order to acquire a shear wave velocity (Vs) to represent liver stiffness.Correlation between Vs and the histological finding was analysed.ResultsAmong 58 successfully modeled rats,9,13,14 and 12 rats were found to be with S1,S2,S3 and S4 of liver fibrosis pathologically,respectively.And 10 rats were found to be with severe inflammatory activity without any fibrosis.Values of Vs increased with the stage of liver fibrosis ( P ＜0.05).There was a significant correlation between Vs and stage of liver fibrosis ( r =0.947,P =0.000).The areas under ROC curve for the diagnosis of fibrosis S≥S1,S≥S2,S≥S3 and S=S4 were 0.983,0.995,0.999 and 0.964,respectively;for the cutoff values of Vs were 1.59 m/s,2.13 m/s,2.33 m/s and 2.51 m/s,respectively,the sensitivity was 95.8％,92.3％,100％ and 84.6％,and specificity was 100％,100％,96.9％ and 95.6％,respectively.The values of Vs in the group with severe inflammatory activity were significantly higher than those in the control group ( P =0.000).ConclusionsARFI has a relatively high value in the evaluation of liver fibrfosis in rats,while severe inflammatory activity may affect its accuracy.

Objective To report a randomized trial in comparing the clinical outcomes of three-port LC versus standard four-port LC. Methods From March 2001 to August 2004, four hundred consecutive patients who underwent elective LC were randomized to receive either the three-port or the four-port technique. All patients were blinded to the type of operation they underwent. Postoperative overall pain and incisional pain at different sites were assessed on the first day after surgery using the Prince-Henry scale. Other outcome measures included length and success of the operation, analgesia requirements, postoperative complications, postoperative stay, and the cosmetic results. Results There was no difference between the two groups in age, sex, weight or other diseases. In terms of outcome, patients in the three-port group had less pain at individual subcostal port sites and better cosmetic results. Success rate, mean operative time, complications, subxiphoid port and overall pain score, analgesia requirements, and postoperative hospital stay were similar between these two groups. Conclusion Three-port LC resulted in less individual port-site pain and similar clinical outcomes but fewer surgical scars compared to four-port LC. The three-port technique is as safe as the standard four-port procedure for LC. Thus, it can be recommended as a routine procedure in elective LC.

Objective To probe the prevention and management of complications after laparoscopic cholecystectomy (LC). Methods Retrospective study was performed on 13 000 patients, who underwent LCs from September 1991 to February 2005 at our department. Results The complication rate was 1. 66% (216 patients) including intraabdominal hemorrhage in 21 patients (0. 16%),bile duct injury in 11 (0. 08% ),gastrointestinal perforation in 7(0. 05% ) , bile leakage in 26(0. 20% ) , retained abdominal tumor in 10(0. 08% ) , retained common bile duct stones in 47(0. 36% ) , intraabdominal abscess in 4(0. 03% ) , upper gastrointestinal hemorrhage in 2(0. 02% ) , extensive subcutaneous emphysema in 32 (0. 25% ) , port wound infection in 46(0. 35% ) , incisional hernia in 1 (0. 01% ) and deep vein thrombosis in 9 (0.07%). Six patients died postoperatively. Conclusions LC is a safe technique when up-to-date equipment and meticulous dissection techniques are employed. With the routine procedure, LC can be performed more safely.

OBJECTIVE To investigate the effects of obstructive sleep apnea hypopnea syndrome on the quality of life of the patients. METHODS The quality of life was assessed with the Medical Outcomes Study SF-36 Health Survey in 145 patients with OSAHS and 17 snorers without OSAHS. Excessive Daytime Sleepiness was assessed with Epworth Sleepiness Scale. Pearson linear correlation and stepwise multiple regression analysis were used to study the association between each domain score of SF-36 and patients information,PSG parameters,EDS scores. RESULTS Three domains of SF-36 correlated negatively with Body Mass Index. Four domains correlated negatively with EDS. Three domains and total score correlated significantly with the lowest oxygen saturation. Two domains correlated negatively with the longest time of apnea. AHI and the lowest oxygen saturation have a mild side effect on the quality of life. BMI and EDS score have significantly side effect on the quality of the life. CONCLUSION Comparing to the mild OSAHS patients and snorers without OSAHS,the severe OSAHS patients have worse quality of life. EDS score and BMI have a significantly side effect on the severe OSAHS patients. OSAHS has a side effect on the quality of life of the patients.

Objective To explore the relationship between 1,25-dihydroxyvitamin D3(1,25-VD3)and alveolar fluid clearance(AFC)in mice in vivo,and investigate its effects in the process of lung fluid clearance. Methods KM male mice were treated with active vitamin D analogue parical-citol(daily i.p. injection)for 2 weeks,and then the in vivo AFC of these mice was measured by bovine serum albumin protein assays. western blot was applied to determine epithelial sodium channel protein levels in lungs of these mice. Results In vivo total AFC was 31.9%±3.8%in vitamin D-treated mice,and significantly lower in the vehicle-treated controls(19.7%±1.9%,P<0.05). Amiloride-sensitive AFC was increased approximate-ly 50%by vitamin D. western blot showed that the expression ofα-epithelial sodium channel was significantly elevated in paricalcitol-treated mouse lungs. Conclusion These observations suggest that vitamin D augments AFC in mice,which may be related to the augment of epithelial sodium channel protein expression. The clinical application of vitamin D therapy may ameliorate pulmonary edema of patients.

OBJECTIVE To evaluate the value of Epworth sleepiness scale(ESS) combined with home-based portable monitoring(PM) in the diagnosis of patients with obstructive sleep apnea syndrome(OSAS). METHODS Adult OSAS patients who visited Beijing Tongren Hospital and complained of snoring were included in this study. The overnight polysomnography(PSG) and the ESS were completed at the sleep center and the portable multi-channel monitoring was completed within one week. Subjects were divided into two groups according to their apnea hypopnea index(AHI) of PSG: simple snoring or mild OSAS group(AHI<15/h) and moderate-severe group(AHI≥15/h). The ESS score, respiratory disturbance index(RDI) of PM and other parameters were compared and the correlation of AHI with ESS scores, RDI were analyzed. The sensitivity and specificity of ESS score≥9 points, RDI≥15/h in the diagnosis of moderate-severe OSAS were calculated. RESULTS A total of 51 subjects were included in this study with 42 males(82.4％) and 9 females(17.6％). Mean age was(43.8±10.8) years, and the mean body mass index was(27.9±4.5)kg/m2; ESS scored 1-24 points and mean(8.6±5.0) points, 18 cases with ESS score ≥9 points; AHI ranged between 2.5 and 99.8/h with a mean of(37.4±29.8)/h. There were 19(37.3％) and 32(62.7％) patients in snoring-mild group and moderate-severe group, respectively. There was significant difference for ESS scores between the two groups(9.9±5.1 vs 6.6±4.2 points, P<0.05),RDI in moderate-severe OSAS group was significantly higher than that in snoring-mild group[(49.4±23.1)/h vs (6.8±4.5)/h, P<0.001)]. There was a positive correlation between the ESS score and PSG derived AHI(r =0.435, P =0.002); the PM derived RDI were significantly correlated with PSG derived AHI in snoring-mild group(r =0.706, P=0.001), moderate-severe group(r =0.873, P=0.000) and patients with ESS score ≥9 points(r =0.967, P=0.000). At a PSG threshold AHI=15/h, the sensitivity and specificity for ESS score ≥9 points were 45.2％ and 78.9％; the PM derived RDI≥15/h had a sensitivity and specificity of 84.4％ and 84.2％, respectively; the sensitivity and specificity for each individual screening tool along with the combination of ESS and PM were 100.0％ and 100.0％. CONCLUSION ESS combined with PM diagnostic method is a valuable measure for the diagnosis of moderate and severe OSAS patients.

Objective To evaluate the effects of different interventios on insulin resistance and ?-cell function in population with impaired glucose tolerance(impaired glucose tolerance,IGT).Methods The total of 382 cases of IGT were divided into five groups according to interventions:control(C),diet plus exercise group(D+E),Acarbose Group(A),Metformin(M)and traditional Chinese medicine Group(TCM),with each group making the appropriate intervention programmes.BMI,FPG,2hPG,F-Ins,P-Ins,F-CP and P-CP were determined before intervention and yearly during the follow up,and BMI,HOMA-IR,HOMA-? were calculated.Results By the end of the study,insulin,F-CP,IR Group D+E and FPG,2hPG,insulin,C-peptide,IR in the other groups were significantly lower than before the intervention(P

The relationship between Human papillomavirus (HPV) 16 infection and the natural course of colorectal adenocarcinoma has not been fully defined. In this study, the HPV 16 E7 DNA was detected in 82 patients with primary colorectal adenocarcinoma to study the relationship between HPV 16 infection and colorectal carcinoma. Samples were taken from both the tumors and the adjacent normal mucosa in the same patient. Polymerase chain reaction (PCR) was used to amplify the HPV16 E7 DNA fragment. The PCR products were gel-purified and sequenced for HPV genotyping. DNA sequence analysis indicated that PCR product was 297 bp. It was the equivalent of 562-858th pairs in the HPV 16 primitive sequences. Our results showed HPV16 E7 DNA expression was significantly higher in colorectal carcinoma (42/82) than in adjacent normal mucosa (4/82). The correlation was found between HPV16 E7 expression and tumor's location; the positive rate was 18.18% in the ascending colon carcinoma group and 64.10% in the rectal carcinoma group. High HPV16 E7 expression was also associated with lower Dukes stage (P < 0.01). These results indicated that there was correlation between colorectal adenocarcinoma and HPV 16 infection. HPV16 infection was relatively higher in the colorectal carcinoma and rare in the adjacent normal mucosa. Specimens expressing higher levels of HPV 16 E7 DNA were associated with lower Dukes stage and more distal location.
Adenocarcinoma ; metabolism ; virology ; Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; metabolism ; virology ; DNA, Viral ; analysis ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Papillomaviridae ; isolation & purification ; Papillomavirus E7 Proteins ; Papillomavirus Infections ; metabolism ; virology