Objective To investigate the clinical application value of fluorescence PCR in vitro diagnosis(IVD)reagent kits in the HLA-B27 detection by comparing 2 kinds of HLA-B27 IVD reagent kit approved by CFDA.Methods A total of 573 clinical blood samples were collected and detected for HLA-B27 by the approved reagent kits based on the fluorescence PCR technique and the flow cytometry.The samples with inconsistent testing results by the two kits were further confirmed by the PCR sequencing.At the same time,about 5% samples of the positive results detected by the fluorescence PCR method were extracted for conducting the re-testing.Results Among 573 samples,191 samples were HLA-B27 positive and 382 cases were HLA-B27 negative by flow cytome-try;the same samples had 194 cases of HLA-B27 positive and 379 cases of HLA-B27 negative by real-time PCR.With flow cytome-try as reference of the final results,the positive coincidence rate of the two kinds of kit was 96.33%(184/191),the negative coinci-dence rate was 94.76%(362/382),27 samples had inconsistent results from the two kinds of assay(accounting for 4.71% of the to-tal number of samples),the total coincidence rate was 95.29% [(184+362)/573],the Kappa value was 0.896(P =0.02);the chi-square test P =0.021,the two kinds of testing method had the high consistency,but the differences existed in the testing results. The re-testing results by PCR sequencing(including 27 samples with inconsistent results by two kinds of kit)were entirely consist-ent with the fluorescence PCR testing results.Conclusion Compared with the authority method flow cytometry for HLA-B27 tes-ting in clinic,the fluorescence PCR kit may present more accurate judging ability for the HLA-B27 testing on the basis of ensuring the higher consistency of the testing results,is easier compared with the sample preparation and operating procedures,and has the stronger clinical application value and prospects s.

Injury of choledocho-pancreatico-duodenal junction refers to the penetrating injury of the bile duct, pancrea-tic duct or duodenal wall in the region of ampulla of Vater. It is often caused by improper operation of surgical instruments, and may lead to leakage of bile, pancreatic or duodenal contents into retroperitoneal space and chemical corrosion in a wide range of retroperitoneal soft tissue, which result in severe secondary infection or even death. Leakage of contrast media, hypertrophy of tissue and anatomical changes were the evidences for injury of choledocho-pancreatico-duodenal junction. Injury of choledocho-pancreatico-duodenal junction can be. divided into 4 types, and treatment selected according to different types of injury is neces-sary for the prognosis of patients.

OBJECTIVE: To observe the influence of Xiaotan Sanjie Recipe on growth of the transplanted tumor and expressions of proliferating cell nuclear antigen (PCNA) and epidermal growth factor receptor (EGFR) in tissue of the gastric carcinoma of nude mice, and to explore the mechanism of Xiaotan Sanjie Recipe in restraining the multiplication of the stomach cancer. METHODS: MKN-45 gastric carcinoma model in nude mice was established. Forty-five nude mice were randomly divided into control group, Xiaotan Sanjie group and 5-fluorouracil (5-FU) group. Drugs were given on the next day of inoculation. Mice in the control group were given normal saline, and mice in the Xiaotan Sanjie group were given Xiaotan Sanjie Recipe. Intraperitoneal injection of 5-FU was administered in the 5-FU group. The expressions of PCNA and EGFR were examined by using streptavidin peroxidase (SP) conjugate technique, and the tumor weight and pathological characteristics were also observed. RESULTS: Compared with the control group, Xiaotan Sanjie Recipe significantly restrained the growth of the transplanted tumor, and reduced the expressions of PCNA and EGFR in tissue of the gastric carcinoma of nude mice (P<0.05). CONCLUSION: Xiaotan Sanjie Recipe can disturb the synthesis of DNA and reduce the proliferous activity of cancer cells by decreasing the expressions of PCNA and EGFR.

Objective To explore the purifying effects of clean bottling workshop in jugged or bottled drinking water manufacturing enterprise in Wuhan. Methods 32 manufacturing enterprises were sampled to test the purifying levels of bottling workshops according to GB50073-2001 The Criterion for Purifying Workshop Design, GB/T16294-1996 Detecting Method for Suspended Particle in Clean Shop or Area of Medical Industry and GB/T16249-1996 Testing Method for Sedimentation Bacteria in Clean Room or Area of Medical Industry. The microorganism determination was based on GB17324-1998 The Hygiene Standard for Bottled Drinking Pure water and GB8537-1995 The Hygiene Standard for Drinking Natural Mineral water and CJ94-1999 The Hygiene Standard for Bottled Drinking Water. Results The qualified rate was 78.1% for the count of dust in air and 81.3% for the count of bacteria in air of bottling workshops of 32 manufacturing enterprises.Among 32 manufacturing enterprises, 25(78.2%) and 8(25%) enterprises accorded with the requirement of 10 000 and 1 000 purifying level of air in whole bottling workshop respectively,only 2 accorded with the requirement of 100 purifying level of air at product line in bottling workshop. The qualified rate was 84.4% for temperature,light,noise,87.5% for flow rate, 46.9% for relative humidity, 34.4% for air pressure difference between indoor and outdoor.After the performance of sanitary management, the qualified rate of microorganism in barrelled water increased from 61.1% in 2001 to 84.1% in 2002. Conclusion The application air cleaning facility in bottling workshop of jugged or bottled drinking water manufacturing enterprise can improve the sanitary quality of microorganism in barrelled water.

Objective To investigate incidence,risk factors and the prevention strategy of local recurrence 6 months after surgery for metastatic bone tumors.Methods Data of 797 patients who had undergone operations for metastatic bone tumors from March 1997 to March 2012 were retrospectively analyzed.Sixty-three patients (7.9％) who had local recurrence 6 months after operation were enrolled in the recurrence group,including 40 males and 23 females,and the average age at the time of operation was 55.21 years.Seven hundred thirty-four patients were enrolled in the non-recurrence group,including 432 males and 302 females,with an average age of 56.49 years.The risk factors for local recurrence 6 months after operation for metastatic bone tumors were statistically analyzed.Results The statistical analysis showed the risk factors for local tumor recurrence 6 months after surgery for metastatic bone tumors included preoperative general condition (10.9％ vs 6.2％),the rate of progress of the primary tumor (10.1％ vs 6.1％),site of bone metastasis (9.1％ vs 3.9％),surgical method (11.4％ vs 6.4％),whether local radiotherapy was performed preoperatively (28.0％ vs 6.6％),whether local radiotherapy was performed postoperatively (8.7％ vs 2.8％),whether sensitive systemic therapy was performed preoperatively (12.2％ vs 6.1％),whether sensitive systemic therapy was performed postoperatively (10.3％ vs 5.6％) and whether local therapy was performed in primary tumor site (10.1％ vs 5.8％).Multivariate analysis showed the independent risk factors included preoperative general condition (OR=0.534),rate of progress of the primary tumor (OR=2.164),site of bone metastasis (OR=2.906),whether local radiotherapy was performed preoperatively (OR=3.184),whether sensitive systemic therapy was performed preoperatively (OR=2.344) and whether sensitive systemic therapy was performed postoperatively (OR =0.468).Conclusion When the patients has following conditions:poor preoperative general condition,fast progressive primary tumor,metastatic tumor in the axial skeleton,application of local radiotherapy preoperatively,and application of sensitive systemic therapy,the surgical treatment should be chosen cautiously.

Objective To evaluate the effect of surgical treatment in facial nerve paralysis.Methods Clinical data of 29 cases in facial nerve paralysis were retrospectively analyzed.All of the 29 cases of facial paralysis,18 cases is the suppurative otitis media,9 cases is the temporal bone fracture,2 cases is the neoplasms of the temporal bone.The 29 cases of facial nerve paralysis were surgical treatment.8 cases by vertical segment or horizontal segment of facial nerve decompression,19 cases by the stylomastoid foramen to the geniculate ganglion of facial nerve decompression,2 cases by the itratemporal course of facial nerve decompression.1 case was underwent end-to-end anastomosis,2 cases of the greater auricular or the sural nerve graft for repairing facial nerve defect.All data were analyzed with Rank sum test.Results Makes a follow-up visit for 6～18 months,the facial nerve function(House-Brackman grading system)before the technique Ⅱ 6.9%,Ⅲ17.2%,Ⅳ34.5%,Ⅴ 31.0%,Ⅵ 10.3%,after the technique,restores Ⅰ 6.9%,Ⅱ27.6%,Ⅲ27.6%,Ⅳ 24.1%,Ⅴ 13.8%,statistics analysis facial nerve function restoreS has the significance difference(P＜0.005).Conclusions The facial nerve decompression and the nerve graft are useful method to treat facial paralysis.Surgical treatment of facial paralysis is satisfied in the suppurative otitis media and the temporal bone fracture.

OBJECTIVE:To observe the clinical efficacy and safety of pregabalin assisting anti-histamine drugs in the treat-ment of secondary skin pruritus in uremic patients underwent maintenance hemodialysis. METHODS:140 uremic patients with sec-ondary skin pruritus underwent maintenance hemodialysis were randomly divided into control group and treatment group,with 70 cases in each group. Control group was given loratadine orally,10 mg,qd. Treatment group was additionally given pregabalin on the basis of control group,75 mg,po,at night of hemodialysis end,increasing by 75 mg every week if no obvious relief was found,maximal dose of 300 mg. Both group received 3 months of treatment. Clinical efficacy,VAG itching score,modified Duo score,PSQI score before and after treatment and the incidence of ADR were compared between 2 groups. RESULTS:Total effec-tive rate of treatment group (90.00%) was significantly higher than that of control group (71.43%),with statistical significance (P<0.05);VAG itching score,modified Duo score and PSQI score of treatment group after treatment were significantly lower than control group and before treatment,with statistical significance(P<0.05). There was no significant difference in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Pregabalin assisting anti-histamine drugs in the treatment of secondary skin pruritus in uremic patients with maintenance hemodialysis can efficiently relieve the itching symptoms,improve sleep quality,and not increase the risk of ADR.

Objective To assess the benefit of intraoperatively placed nasointestinal feeding tubes for early postoperative enteral nutrition.Methods The retrospective study was carried out on 19 cases of in-fants with long gap esophageal atresia who underwent one-stage operation in Xinhua Hospital from January 2006 to December 2013.Patients were divided into two groups according to whether the transanastomotic na-sointestinal feeding tubes(TNFT)were placed.Demographics,associated anomalies,operative findings,com-plications,duration of enteral nutrition(EN),length of stay were analyzed.Results Thirty-seven patients had long gap esophageal atresia and nineteen underwent one-stage operation.Eight patients were identified as TNFT group while eleven patients were identified as nonfeeding tube(NFT)group.There was no significant differences between the two groups in gestational age,birth weight,associated anomalies,anastomotic leak, weight gain,ICU-free days and length of stay(P>0.05).But the ventilator-free days(23.5 d vs.19.3 d), first EN time(0.8 d vs.9.1 d)and total EN feeding time(15.9 d vs.21.4 d)between the two groups were significantly different(P<0.05 ).Conclusion Intraoperatively placed TNFT may lead to earlier EN feeding and shorter parenteral nutrition duration,but a larger prospective study would be required to prove the benefits and ensure the safety.

Objective To analyze the immunophenotyping characteristics of myeloid leukemia in transgenic mouse.Methods According to differential antigen expression profile on various hematopoietic lineages,flow cytometric analysis of bone marrow sample was performed on 5 myeloid leukemia mice and 10 healthy BL6 mice.Cell cycle analysis was further performed to assess cell proliferation.Results Expressions of Mac-1+ Gr-1+ and c-Kit+ in bone marrow cells in transgenic leukemia mice were(72.6±6.5)% and (20.5±4.8)%,and it were significantly higher than those in normal mice[(52.8±4.8)% and(2.1±0.3)%](t=6.66,12.66,P＜0.01).And expressions of B220+,CD3+,CD41+ and Ter119+ in leukemia mice were(2.7±1.1)%,(1.2±0.3)%,(1.2±0.6)% and(2.8±1.1)%,respectively.It were significantly lower than those in normal mice[(20.2±2.1)%.(6.6±1.3)%,(4.7±1.1)% and(10.6±1.2)%](t=-17.63,-8.69,-6.30,-12.28,P＜0.01).The percentages of S phase and G2/M phase in leukemia mice were(25.7±4.2)% and(21.1±4.2)%,respectively.It were significantly increased as compared with normal mice[(11.8±2.1)% and(8.9±1.8)%](t=8.59,7.98,P＜0.01).Conclusions Immunophenotyping of myeloid leukemia in transgenic mouse was characterized by hish expression of myeloid specific marker(Mac-1 and Gr-1)and hematopoietic stem/progenitors cells specific marker(c-Kit),and by low expression of B-lymphoid specific marker(B220),T-lymphoid(CD3),megakaryocyte(CD41)and Erythroid(Ter119).

Objective To analyze the diagnostic process of a rare case of acute myeloid leukemia with minimal differentiation undergoing a lineage switch to mixed phenotype acute leukemia, NOS-rare types,and to investigate its difference from other acute myeloid leukemia and mixed phenotype acute leukemia. Methods Following tests were performed on the patient with switched mixed phenotype acute leukemia and three control leukemia patients ( including two acute myeloid leukemia with minimal differentiation and one mixed phenotype acute leukemia ). Cell morphology was analyzed by bone marrow smear and related cell chemical staining. Immunophenotyping of bone marrow was performed by flow cytometry ( FCM ). G-banding technique was used for karyotype analysis and RT-PCR was used for fusion gene detection. All the laboratory data of the switched patient were compared to that of three control patients in order to reveal the characteristics of such a rare phenotype switch in acute leukemia. Results Before switching, the morphology of acute myeloid leukemia with minimal differentiation demonstrated 0.82 blasts occurring in bone marrow, distinct nucleoli and absence of Auer rods. Blast cells expressed hematopoieticassociated antigens ( CD38, HLA-DR ), myeloid antigens ( CD13, CD56, CD11b ) and CD7. And these blasts were negative for MPO, CD33, CD15, CD79, CD19, CD22, cytoplasmic CD3, CD4 and CD8. After switching, 0. 42 blasts were found in bone marrow, showed eosinophilia and presence of basophile. Blast cells expressed hematopoietic-associated antigens ( CD38, HLA-DR ), myeloid antigens ( MPO, CD13 ),lymphoid antigens ( CD19, CD79a ,cytoplasmic CD3, and CD7 ). The control group showed typical morphology and immunophenotyping. No abnormal karyotype and fusion gene were detected. Conclusions It is a rare and complicated case that acute myeloid leukemia with minimal differentiation switched to mixed phenotype acute leukemia, NOS-rare types. The laboratory features, especially the change of immunophenotyping play an important role in the diagnosis.