Objective To investigated the percutaneous transhepatic gallbladder drainage combined laparoscopic cholecystectomy in the treatment of purulent cholecystolithiasis for serious patients.Methods Our hospital in the past two years,76 underwent lap-aroscopic cholecystectomy in the acute calculous suppurative cholecystitis of elderly patients with clinical data were retrospectively analyzed.Patients could be divided into group A(surgery)within 72 hours after admission,group B(conservative treatment after percutaneous puncture drainage of the gallbladder,again)elective surgical procedures,A total of 2 groups.Comparative analysis of two groups of patients with complications of surgery,laparotomy rate and total effective rate.Results A,B two groups of postoper-ative complication rates were 36.11% and 15.00% respectively,each postoperative complication rates had significant difference(P<0.05);A,B two groups of transit open rate was 13.89% and 15.00%,A,B two groups of operation time,transit open rate had no significant difference(P>0.05),the total effective rate was lower than that in group B,group A statistically significant differ-ence(P<0.05).Conclusion PTGD combined LC is simple and effective treatment in high-risk acute purulent cholecystolithiasis.

Objective To explore the role of insulin-like growth factor-2/insulin-like growth factor1 receptor/insulin receptor substrate-1 (IGF2/IGF1R/IRS1) signal pathway inducing the chemoresistance of epidermal growth factor receptor 2 (ErbB2) positive breast cancer cells to Herceptin.Methods Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot assay were used to determine the expression levels of IGF2,IGF1 R,and IRS1.The direct targets of miR-126 were validated by dual-luciferase reporter gene assay.In SKBR3/pool2 cells,IGF1 R activity was reduced by an inhibitor of IGF1 R,and IRS1 was knocked-down by shRNAs.Furthermore,3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed to evaluate the sensitivity of these treated cells to Herceptin.Results IGF2,IGF1 R,and IRS1 were significantly higher expressed in SKBR3/pool2 cell compared to that in SKBR3 cell.Western blot assay showed that IGF2/IGF1R/IRS1 was activated in SKBR3/pool2 cells.Bioinformatics analysis combined with luciferase activity suggested that miR-126 directly targeted IRS1.MTS results demonstrated that the chemosensitivity to Herceptin of SKBR3/ pool2 cells with inhibitor of IGF1R or shRNAs targeting IRS1 or overexpressing miR-126 was significantly reduced.Conclusions IGF2/IGF1R/IRS1 signal pathway confers to the chemoresistance of ErbB2 positive breast cancer cells to Herceptin.

] Objective To explore the role and mechanisms of FGF2 in chemo?resistance in breast cancer. Methods The inhibitors for different signal pathway were used to treat two drug?resistant breast cancer cell lines MCF?7/5?Fu and T47D/5?Fu established in our lab. MTS assay was used to determine chemo?sensitivity and Hoechst stain was used to measure apoptosis. Protein activation and FGF2 protein level in cell culture medium were detected by western blot and ELISA respectively. Results Akt inhibitor MK?2206 (20 nM) and mTOR inhibitor AZD8055 (2 nM) significantly reversed the chemo?resistance of MCF?7/5?Fu and T47D/5?Fu cell lines to 5?Fu and paclitaxel, but ERK1/2 inhibitor SCH772984 showed no significant effect. Compared to parent cell lines MCF?7 and T47D, p?Akt and p?S6K (represented as mTORactivity) levels were obviously up?regulated in MCF?7/5?Fu and T47D/5?Fu cell lines, and so do the FGF2 mRNA level and FGF2 protein level from culture medium. Moreover, FGFR inhibitor AZD4547 (4 nM) markedly reversed the chemo?resistance of MCF?7/5?Fu and T47D/5?Fu cell lines to 5?Fu and paclitaxel and down?regulated activation of FGFR?Akt?mTOR signal pathway. In agreement, FGF2 protein (10ng/ml) enhanced the chemo?resistance of MCF?7 and T47D cell lines to 5?Fu and paclitaxel and up?regulated activation of FGFR?Akt?mTOR signal pathway. Conclusion Activation of FGF2?FGFR?Akt?mTOR signal pathway promoted chemo?resistance of breast cancer cells.

Objective:To characterize the memory B lymphocytes activalion by nonspecific antigen of different phylogeny antigen.Methods:Three groups were immunized by same quantity different antigens,which were ovalbumin,milk and ostrcean extract to intitule A,B,C,respectively,as basic immunity.After one week,across immunity was performed with A,B and C antigen as secondary immunity,the subgroups were divided.Basic antibodies were determined with ELISA before and after secondary immunity.Results:Antibodies after secondary immunity increase 10%～25% than that before secondary immunity in subgroups which antigens were different when basic and secondary immunity.There were significant difference compare with control groups (P

Objective To investigate the low hemoglobin density (low haemoglobin density,LHD) and red blood cell volume distribution width (red blood cell volume distribution width,RDW) in iron deficiency anemia (iron-deficiency anemia,IDA)in children with the value of diagnosis and treatment.Methods From February 2016 to May 2017,in the Second People's Hospital of Longgang District in Shenzhen City,86 cases of children with iron deficiency anemia for IDA group,and 120 cases of healthy children (as the control group) at the same time were confirmed.Blood routine of children with IDA were detected before and after treatment hemoglobin concentration and the results were analyzed statistically.Results 120 cases of healthy children in the peripheral blood LHD value was 2.74 ％ ± 0.90 ％ and the boy was 3.07 ％ ± 0.81％,higher than that of 2.26 ％ ± 0.69 ％ of the girls.Between them,there was statistically significant difference (t=3.815,P＜0.05).The value of RDW was 12.37 ％ ± 2.84 ％,12.09 ％ ± 2.80 ％ for the boys and 12.56 ％ ± 2.87 ％ for the girls,there was no statistically significant difference between the them (t=0.517,P＞0.517).Before treatment,86 cases of children with IDA LHD and RDW values in peripheral blood were 30.67 ％ ± 20.12 ％ and 16.62 ％ ± 3.27 ％,significantly higher than the control group,the difference was statistically significant between (t=4.025 ～ 4.025,P＜0.05 ～ 0.01),and there was no statistically significant difference between male and female children (t =0.761 ～ 0.917,P＞ 0.05).After treatment L HD and RDW values were 10.65 ％ ± 8.92 ％ and 14.02 ％ ± 2.93 ％,significantlylower than before the treatment,between them was statistically significant difference (t=2.912～9.675,P＜0.05).Conclusion Children with iron deficiency anemia treatment before the LHD and RDW values significantly elevated in the blood,significantly decreased after treatment,the LHD and RDW values for diagnosis and treatment of children with iron deficiency anemia dynamic monitoring has certain application value,worthy of popularization and application.

Objective To sequence and analyze the envelope (E) gene of type Ⅰ dengue virus isolated from Guangzhou in 2009 for tracing the infection source. Methods The serum samples were collected from patients diagnosed with dengue fever in Guangzhou area during 2009. Dengue virus was isolated and cultured in C6/36 cells.The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and then sequenced. The phylogenetic tree was drawn by neighbor-joining method. The bioinformatics analysis was performed by combining the phylogenetic information and the epidemiology data. Results Four strains of type Ⅰ dengue virus were isolated from 19 samples. E gene of these strains was amplified and sequenced. The phylogenetic analysis showed that 09/GZ/9104 strain and 09/GZ/9236 strain had identical nucleotide sequence and fell within the American/African group, 09/GZ/11534 stain and 09/GZ/11562 strain had similar sequence homology and fell within the Asian group. Conclusion The typeⅠdengue viruses in Guangzhou area in 2009 are imported, which belong to two genotypes and may come from two independent origins respectively.

Objective To measure serum surfactant protein (SP) A and D levels in patients with interstitial lung disease associated with rheumatoid arthritis (RA).Methods Serum SP-A and SP-D levels of RA,RA-ILD patients and healthy controls were assessed using a sensitive enzyme-linked immunosorbent assay (ELISA).The relationship between SP-A and SP-D and RA-ILD was analyzed.The serum SP-A and SP-Dpositive rate was calculated for the three groups.The correlation between SP-A and SP-D with RF,anti-CCP,antinuclear antibody,antikeratin antibody,anti-perinuclear factor,C-reactive protein,erythrocyte sedimentation rate,were analyzed.Mean value of groups were compared with variance analysis,Spearmam rank correlation test was used for correlation analysis.Results The levels of serum SP-A in RA-ILD patients and RA patients as well as in healthy controls were [ (51.2±9.2),(25.9±2.6),( 15A±0.3 ) μg/L] respectively.The level of serum SP-D of the three groups was [ ( 42.5 ±8.1 ),(20.8 ± 1.5 ),( 16.6±0.8 ) μg/L ] respectively.The levels of serum SP-A and SP-D in patients with RA complicated with ILD were higher than those simple RA patients and healthy controls (P＜0.05).The levels of serum SP-A and SP-D in patients with RA were not significantly higher than those in healthy controls (P＞0.05).The positive rate of serum SP-A and SP-D in RA-ILD patients were significantly higher than those in simple RA patients and healthy controls.The positive rate of serum SP-D of RA-ILD patients was higher than that of SP-A.The levels of serum SP-A and SP-D in patients with RA complicated with ILD were correlated positively with age,C-reactive protein.The level of serum SP-D was correlated positively with RF,anti-CCP,antikeratin antibody.There was no correlation between the level of serum SP-A and SP-D with RA-ILD and antinuclear antibody,antiperinuclear factor,erythrocyte sedimentation rate.Conclusion The levels of serum SP-A and SP-D are correlated with RA-ILD and may be useful markers for ILD in patients with RA.These two paramenters may be helpful to early diagnosis of RA-ILD.The Serum SP-D levels are more sensitive in predicting the development of RA-ILD than other parameters and can help in assessing the severity of lung damage.

Objective To compare the significance of DNA-associated autoantibodies to cell membrane(cmDNA)in systemic lupus erythematosus(SLE)detected with indirect immunofluorescence on human B lymphoma cell line Raji and pmmyelocytic line HL60.Methods Indirect immunofluorescence assay both on cell line Raji and HL60 was used to measure anti-cmDNA antibodies in sera of 306 SLE patients.192 patients with other rheumatic diseases and 50 healthy controls.Results Indirect immunofluorescence assay on cell line Raji was used to measure anti-cmDNA antibodies.72.5% SLE and 10.4% other rheumatic diseases were positive for anti-cmDNA,but negative in 50 blood donors(P<0.01).Indirect immunofluorescence assay on cell line HL60 was used to measure anti-cmDNA antibodies,76.1% SLE and 16.7% other rheumatic diseases were positive for anti-cmDNA,but negative in 50 blood donors(P<0.01).The sensitivity of anti-cmDNA were 72.5%and 76.1%,respectively.The specificity of anti-cmDNA was 91.7% and 86.8%,respectively.There was no significant difference in sensitivity and spocificity(P>0.05).The methods of culture,freeze and resuscitation on the two cells were similar.but cell line Raji was easier to resuscitate than cell line HL60.Observing with fluorescence microscope.we find that cmDNA was expressed on the both cells and the staining was stronger on cellline Raji than HL60.Conclusion Anti-cmDNA antibody has high positivity which is one of the most valuable marker in the diagnosis of SLE.We recommend to measure anti-cmDNA antibodies with indirect immunofluorescence assay on cell line Raji rather than HL60.

ObjectiveTo compare the significance of anti-cmDNA antibody in systemic lupus erythematosus (SLE) patients detected with IIF on human's B lymphoma cell line Raji and promyelocytic line HL60.The diagnostic value of anti-cmDNA antibody in SLE was also explored.MethodsThree hundred and six patients with SLE were included in this study.As control groups,we included 192 patients with other rheumatic diseases and 50 healthy controls.The testing method for anti-cmDNA antibody was set up.The assessment of the significance of anti-cmDNA antibody in SLE detected with IIF on cell line Raji and HL60 was carried out andthe diagnostic value of anti-cmDNA antibody in SLE was investigated.ANA and antidsDNA antibody were measured by IIF at the same time.Anti-Sm was measured by immuno-diffusion andWestern blotting.AnuA was tested by enzyme linked immunosorbent assay.The statistical methods used in this study including McNemar X2 test,Spearman related test and Logistic regression analysis.Results The fluorescence brightness of Raji cell line was stronger than HL60 cell line.There was no statistically significant difference in the sensitivity and specificity of anti-cmDNA antibody in SLE detected with IIF with Raji or HL60 cell lines (P＞0.05).The sensitivity of anti-cmDNA antibody detected with IIF on Raji cell line was higher than anti-dsDNA antibody and anti-Sm antibody(P＜0.01),while the specificity of anti-cmDNA antibody was similar to anti-dsDNA antibody (P＞0.05) and was lower than anti-Sin antibody (P＜0.01).The sensitivity of anti-cmDNA antibody was similar to AnuA(P＞0.05) and the specificity was lower than AnuA (P＜0.01).The sensitivity of ANA was higher than anti-cmDNA antibody (P＜0.01) and the specificity was much lower than anti-cmDNA antibody(P＜0.01).The sensitivities of anti-dsDNA antibody,anti-Sm antibody and AnuA were much higher when combined with anti-dsDNA antibody than any one antibody only (P＜0.05).Anti-cmDNA antibody was correlated with mucosa ulcer in SLE patients(OR=2.343,P=0.029).The ESR of SLE patients was also correlated with anti-cmDNA antibody(OR=l.031,P=0.012).Anti-cmDNA antibody was not correlated with SLEDAI (r=0.070,P=0.600).ConclusionRaji cell line is better than HL60 cell line in detecting anti-cmDNA antibody with IIF.Anti-cmDNA antibody has higher sensitivity and specificity in SLE.Combined detection of anti-cmDNA antibody and other autoantibodies can further improve the diagnostic accuracy of SLE.

Purpose To evaluate the value of real-time contrast-enhanced ultrasound (CEUS) combined with comparative observation in determining the initial time of peripheral lung lesion,and to provide a new method for the diagnosis of peripheral lung disease by CEUS.Materials and Methods Sixty patients with peripheral lung disease diagnosed by chest X-ray or CT examination were examined by CEUS,and the lesions and perfusion of surrounding lung tissue were observed in real time and comparatively.The time of initial enhancement of peripheral lung lesion was determined by real-time comparative observation method,that is,to compare the time of contrast agent to reach the peripheral lung lesion with the time to reach the adjacent gas-containing lung tissue.All lesion specimens were obtained by surgery or ultrasound-guided percutaneous biopsy,and the pathological diagnosis was performed.According to the pathological results,the peripheral lung lesions were classified into the malignant group,the chronic inflammation group and the pneumonia group.Results Sixty cases of peripheral lung lesions were identified as 38 cases of malignant tumors (including 15 cases ofsquamous cell carcinomas,9 cases of adenocarcinoma cell carcinomas,2 cases of large cell carcinomas,9 cases of small cell carcinomas and 3 cases of metastatic carcinomas),14 cases of chronic inflammation lesions (including 7 cases of tuberculomas,3 cases of inflammatory pseudotumors and 4 cases of lung abscess) and 8 cases of pneumonia.The detection rates of pulmonary arterial phase and bronchial artery phase using the real-time comparative observation method were 100％ (60/60) and 85％ (51/60),respectively.The initial time of pulmonary arterial phase and bronchial artery phase were (6.1 ± 0.9) s and (10.5 ± 1.6) s,respectively.The initial time of pulmonary arterial phase was sooner than bronchial artery phase,the difference was statistical significance (P＜0.05).Conclusion The real-time CEUS combined with comparative observation method has important diagnostic value in judging the initial enhancement time of peripheral lung lesion and can provide a new method for the accurate diagnosis of peripheral lung disease,which is worth popularizing.