ObjectiveTo develop an assay to quantitatively detect anti-BP180NC16A IgG subclasses and to assess the significance of anti-BP180NC16A lgG in bullous pemphigoid (BP).MethodsThe Glutathione S-transferase(GST)-BPI80NC16A fusion protein was expressed in E.coli system and purified by affinity chromatography.An improved enzyme-linked immunoabsorbent assay (ELISA) was developed and used to detect anti-BP180NCI6A IgG subclasses in serum samples from 10 patients with BP,5 patients with pemphigoid gestationis (PG),1 patient with linear IgA bullous dermatosis (LIBD) and 2 patients with pemphigus.Results The optimal condition for the ELISA was determined by cross assay as follows:the concentration of GSTBP180NC16A fusion protein for coating,500 μg/L; the condition for coating,4 ℃ for 12 hours; the dilution ratio of sera and secondary antibody,1∶ 100 and 1 ∶ 2000 respectively; the condition for incubation,37 ℃ for 1 hour;,the condition for the enzyme-substrate reaction,37 ℃ for 20 minutes.Of the 10 patients with BP,all were positive for anti-BP180NC16A IgG1,9 for IgG2,5 for IgG3,and 9 for IgG4.Anti-BP180NC16A IgG was undetected in any of the serum samples from 2 patients with pemphigus vulgaris or 1 patient with adult LIBD.All the 5 sera from patients with PG; were positive for all the anti-BP180NC16A IgG subclasses,which were predominated by IgGl and IgG3.ConclusionThe developed ELISA is a highly specific and reproducible semiquantitative method for the detection of anti-BP180NCI6A IgG subclasses in patients with BP and PG.

Objective To explore the clinical value of enhanced recovery after surgery (ERAS) in the perioperative period of biliary surgery.Methods The prospective study was adopted.The clinical data of 800 patients diagnosed as benign biliary tract diseases who were admitted to the Cangzhou's Central Hospital from March 2011 to January 2015 were collected.Patients were randomly divided into the case and control groups by random sequence method.The patients of case group received the ERAS management and patients of the control group received traditional perioperative management.(1) Stress state evaluation:the insulin resistance (IR) index and level of serum C-reactive protein (CRP) were calculated.(2) Levels of inflammatory cytokines:levels of IL-6 and TNF-α in the 2 groups were detected by enzyme-linked immunosorbent assay (ELISA).(3)Nutritional status:level of serum prealbumin (PA) was detected.(4) Postoperative status:time to anal exsufflation,time of defecation,duration of hospital stay and treatment expenses were recorded.(5) Postoperative complications were observed.Measurement data with normal distribution were represented as x ± s and comparison between groups was analyzed using t test.Repeated measurement data were analyzed by the repeated measures ANOVA.Count data were represented as ratio and analyzed using the chi-square test.Results The 800 patients were screened for eligibility,including 400 in each group.(1) Stress state evaluation:the levels of IR and CRP from pre-operation to 7 days after operation were from 3.8 ± 0.8 to 3.7 ± 0.7 and from (18 ± 5) μg/L to (27 ±8) μg/L in the case group,from 3.9 ±0.9 to 3.8 ±1.0 and from (18 ±5) μg/L to (38 ±9) μg/L in the control group,respectively,with statistically significant differences in the changing trends (F =12.38,17.85,P ＜0.05).(2) Comparisons of levels of inflammatory cytokines:levels of IL-6 and TNF-α from pre-operation to 7 days after operation were from (9.3 ± 3.1) ng/L to (12.5 ± 2.8) ng/L and from (10.2 ± 3.5) ng/L to (12.8 ±3.3)ng/L in the case group,from (8.9 ±3.3)ng/L to (17.3 ±6.5)ng/L and from (9.9 ±2.8) ng/L to (14.3 ±4.2)ng/L in the control group,with statistically significant differences in the changing trends (F =19.93,15.74,P ＜ 0.05).(3) Comparison of nutritional status:level of PA from pre-operation to 7 days after operation was from (335 ± 53)mg/L to (332 ±50)mg/L in the case group and from (330 ± 49)mg/L to (331 ± 45)mg/L in the control group,with a statistically significant difference in the changing trends (F =4.46,P ＜ 0.05).(4)Comparisons of postoperative status:time to anal exsufflation,time of defecation,duration of hospital stay and treatment expenses were (30 ± 10)hours,(51 ± 13) hours,(7.7 ± 2.5) days,(10.2 ±2.3) × 103 yuan in the case group and (54 ± 8) hours,(70 ± 16) hours,(15.4 ± 3.1) days,(15.6 ± 4.7) × 103 yuan in the control group,with statistically significant differences (t =37.73,18.62,38.67,20.64,P ＜0.05).(5) Comparisons of postoperative complications:incidence of complications was 15.00％ (60/400)in the case group and 22.50％(90/400) in the control group,and the numbers of patients with incision infection,nausea and vomiting,abdominal infection,intra-abdominal infection,urinary tract infection,intro-abdominal hemorrhage and bile leakage were 23,15,8,5,4,3,2 in the case group and 30,20,13,10,7,5,5 in the control group,respectively,showing statistically significant differences in the incidence of complications (x2=7.39,P ＜ 0.05).Conclusion ERAS management in the perioperative period of biliary surgery is beneficial to postoperative recovery of patients,and it can also relieve postoperative stress state and inflammatory response,reduce the duration of hospital stay,treatment expenses and incidence of postoperative complications.

Bullous pemphigoid (BP) is a group of autoimmune blistering skin diseases.There are special measures for the assessment of disease severity and therapeutic response in autoimmune bullous skin diseases.Common scoring indexes include autoimmune bullous skin disorder intensity score (ABSIS),bullous pemphigoid disease area index (BPDAI),and so on.ABSIS has been validated,and is widely applied in evaluation of pemphigus vulgaris and other autoimmune blistering diseases.BPDAI is still in the process of verification,and has been applied in only a few clinical trials.The evaluation of diseases by ABSIS and BPDAI is both based on a global assessment of skin and mucosal lesions,and degree of subjective pruritus.

Objective:To establish a method to determine ferrous succinate in ferrous succinate tablets .Methods:Ion chromatog-raphy was performed on an IonPac AG19(4 mm ×250 mm)anion exchange chromatography column with a guard column of Ion Pac TM AG19( 4 mm ×50 mm);30.00 mmol· L-1 KOH was used as the isocratic eluent at a flow rate of 1.00 ml· min-1 with suppressed conductivity detection ( Curb:75 mA) at the temperature of 30℃.The sample size was 25 μl .Results:The linear range of succinic acid was 2.005 1-25.064 0μg· ml-1(r=0.999 5).The detection limit and quantitation limit was 0.12ng and 0.42ng, respectively. The recovery was 101.45%(RSD=0.43%, n=6).Conclusion:The method is simple, rapid, accurate, reproducible and sensitive , and can be used to determine the ferrous succinate content in ferrous succinate tablets .

Objective:To establish a rapid HPLC testing method for chloroquine phosphate,hydroxychloroquine sulfate and amodiaquine hydrochloride.Methods:The chromatographic separation was performed on a GRACE prevail C18(53 mm×7 mm,3 μm)column,and the column temperature was maintained at 30℃.Acetonitrile-0.3% triethylamine acetonitrile solution (adjusting pH to 3.0 with phosphoric acid) (12∶88) was used as the mobile phase,the flow rate was 1.0 ml· min-1 and the UV detection wavelength was 254 nm.The qualitative research was performed using relative retention time and spectral similarity as the double indicators.The relative correction factor in the quantification analysis was used for the content determination.Results:Three anti-malarial drugs showed good behavior in one chromatographic system.The rapid HPLC testing analysis could be achieved.The qualitative research was more accurate by using the double indicators (UV spectral similarity and relative capacity factor).The HPLC qualitative accuracy was increased.The relative correction factor method for the quantification could effectively reduce the use of reference substances and speed up the analysis of HPLC.Conclusion:The method is rapid and simple,and suitable for the rapid determination of drugs.

Objective To evaluate the safety and efficacy of one-stage endoscopic sphincterotomy plus laparoscopic cholecystectomy for the treatment of patients who have concurrent gallstones and CBD stones.Methods We conducted a prospective study from Mar 2008 to Nov 2011 in our department.A total of 63 consecutive patients of concurrent gallstones and CBD stones were included and divided into two groups:31 patients in test group first underwent ERCP and EST to remove CBD stones,and then during the same general anesthesia underwent laparoscopic cholecystectomy (LC) ; while 32 patients in control group first underwent ERCP and ES,and then LC 3-5 days later.Results The stone clearance rate in test group and control group were 96.9％ and 96.7％ (P ＞ 0.05),respectively.The postoperative lung infection rate in the test group and the control group were 7.1％ and 16.1％ (P ＜0.05),respectively.The length of hospital stay (LOS) of the two groups was (7.5 ± 1.7) days and (12.6 ±2.5) days,respectively (P ＜ 0.05).The total expense of hospitalization was (￥) (2 356 ± 126) and (￥) (37 056 ± 152),respectively (P ＜ 0.05).Conclusions One-stage laparoendoscopic management of patients with concurrent gallstones and CBD stones is safe,effective and more economic approach than two-stage treatment for patients suffering from concurrent gall stone and choledocholithiasis.

Objective To measure the expressions of IL-10, IL-23 and CD86 in lesions of epidermodysplasia verruciformis (EV), and to explore the relationship between cellular immune abnormality and EV pathogenesis. Methods Immunohistochemistry was used to measure the expressions of IL-10, IL-23 and CD86 in tissue samples from 10 patients with EV and 10 normal human controls. Results Three cytokines were observed in all the samples of EV, with the expression score ranging from 3 to 6 and expression intensity from moderate to high. However, of the control specimens, only 1 was positive for IL-10 with the expression score being 3, and expression intensity being moderate. Conclusion The pathogenesis of epidermodysplasia verruciformis may be correlated with the expression abnormality of some cytokines secreted by keratinocytes.

Objective To study the function of Contour stapler in the sphincter-preserving surgery in 121 cases with low rectal cancer. Methods One hundred and twenty-one cases enrolled were divided into 2 groups randomly and accepted Contour or other kinds of staplers after operation respectively. Further comparison were performed on the rate of sphincter-preserving,anastomosis stricture and leakage,infection of incision and stool frequency between two groups. Results We found no significant differences in the comparison of rate of anastomosis stricture or leakage,stool frequency,but the Contour group had significant higher rate of sphincter-preserving (98. 3%)than other stapler group(68. 9%),and lower rate of infection of incision(3.94%)than control(11.9%)(P ＜0. 05 respectively). Conclusions It is secure,reliable and practical to use Contour stapler in operation on low rectal cancer,which can increase the opportunity of sphincter-preserving,decrease the the rate of infection of incision.

Objective To explore the correlation between ATF5 protein expression level and drug sensitivity of gastric cancer AGS cells.Methods We established the high,middle and low ATF5 expression groups by transfecting the AGS cells with ATF5 pcDNA3.1 plasmid,ATF5 siRNA and empty plasmid using the liposome transfection technique.Then we detected the expression of the ATF5 protein of the 3 groups using the Western Blot.The drug sensitivity of AGS cells in 3 groups to paclitaxel and cisplatin was detected using the MTT experiments and plate clone formation experiments then the correlation between ATF5 protein expression level and drug sensitivity of gastric cancer AGS cells was analyzed.Results With the effect of paclitaxel or cisplatin on AGS cells in 3 groups,the surviving fraction and the IC50 of high ATF5 expression group were higher than those of control group,which indicated statistical significance (all P ＜ 0.05);while the surviving fraction and the IC50 of low ATF5 expression group were lower than those of control group (all P ＜ 0.05,P cisplatin =0.00,P paclitaxel =0.002).We found that cell clone formation of AGS cells to paclitaxel and cisplatin was significantly increased after the transfection with ATF5 protein plasmid and decreased after transfection with ATF5 siRNA plasmid (all P ＜ 0.05).Conclusion The expression of ATF5 protein is related to the drug sensitivity of gastric cancer AGS cells that upregulation of ATF5 protein expression can decrease the drug sensitivity of AGS cells to paclitaxel and cisplatin in gastric cancer.

Objective To study the possible relationship between human papillomaviruse (HPV) infection and external genital proliferative diseases. Methods HPV DNA was detected using polymerase chain reaction (PCR) with a consensus primer and typed by using restriction fragment length polymorphism (RFLP) method in 151 paraffin-embedded biopsy specimens from seven kinds of external genital epidermal proliferative lesions. Results In 30 cases of condyloma acuminatum, HPV DNA was positive for all cases, among which HPV6 and HPV11 accounted for 60% and 26.7%, respectively. In 40 cases of bowenoid papulosis, 5 cases of genital Bowen′s disease and 6 cases of erythroplasia of Queyrat, the positive rates of HPV DNA were 55%, 100% and 33.3%, respectively, and the predominant type was HPV16. In 18 cases of external genital invasive squamous cell carcinoma HPV DNA positive rate was 27.8%, and HPV16 was detected in all of the 5 HPV-positive cases. HPV DNA was negative in 32 cases of leukoplakia and 20 cases of extramammary Paget′s disease. Conclusions Condyloma acuminatum is mainly caused by HPV6 infection, followed by HPV11; HPV16 infection is closely associated with the pathogenesis of bowenoid papulosis and external genital squmous cell carcinoma including Bowen′s disease and erythroplasia of Queyrat. In external genital invasive squamous cell carcinoma, HPV16 infection may be one of the multiple carcinogenic factors. No clear relationship is found between HPV infection and leukoplakia or extramammary Paget′s disease.