Objective To analyze the drug resistance status of mycobacterium tuberculosis in patients with double immunization of human immunodeficiency virus (HIV) and tuberculosis (TB) by phage bioassay (PhaB),and to optimize the control strategy.Methods One hundred and twelve cases of HIV/TB infected patients.in Chongqing Ninth People's Hospital were treated with PhaB method,and the drug susceptibility testing results were compared with 208 cases of simple pulmonary tuberculosis patients.Results The anti-tuberculosis drug resistance rate of HIV/TB patients was lower than that of simple pulmonary tuberculosis patients.The resistance rates of 5 common anti-tuberculosis drugs in HIV/TB patients were 7.14％ of isoniazid (INH),7.14％ of pyrazinamide (PZA),5.36 ％ of rifampicin(RFP) streptomycin(SM),and 4.46 ％ of ethambutol (EMB),compared with simple pulmonary tuberculosis(resistance rates of RFP were 17.31％,IN H 13.46 ％,PZA 11.54 ％,EMB 10.58 ％,SM 9.62 ％),RFP resistance rate of HIV/TB infected patients was lower(P＜0.05).There was no significant difference between two groups in the other four anti-tuberculosis drug(P＞0.05).The coincidence rate with the absolute concentration method were INH 96.4％,RFP 98.2％,PZA 96.4％,EMB 93.8％ and SM 96.4％,respectively.Conclusion The resistance rate of mycobacterium tuberculosis to RFP in patients with HIV/TB infection in this region is lower than that in patients with common pulmonary tuberculosis,which is related to the good medication compliance of these patients.PhaB has the characteristic of fast,simple,without special equipment,it can be used as a rapid screening of mycobacterium tuberculosis drug resistance method.

Objective To evaluate the relationship between the amount of grafted cells and the incidence of acute graft versus host disease (aGVHD) after haploid hematopoietic stem cell transplantation (haplo-HSCT). Methods Data of 68 patients who underwent haplo-HSCT from Jan 2009 to Dec 2013 were analyzed retrospectively. Influences of different factors on the incidence of Ⅲ-Ⅳ degree of aGVHD after HSCT were evaluated. Results 68 patients including 42 males and 26 females were 5/10-9/10 HLA match with 19 father donors, 24 mother donors, 16 sibling donors and 9 children donors. 51 patients not suffered Ⅲ-Ⅳdegree of aGVHD included 32 males and 19 females with the mean age of 20 years old (5-55 years old). 17 patients sufferedⅢ-Ⅳdegree of aGVHD including 10 males and 7 females with the mean age of 23 years old (5-54 years old). There were no significant differences in the amount of the grafted mononuclear cells (MNC) and CD34+cells, and the white blood cell counts (WBC) and platelet count (Plt) recovered time between two groups (P>0.05). However, MNC number was related to CD34+cell number (P<0.05) and WBC recover time (P<0.05), and the CD34+cells number was related to WBC and Plt recover time (P< 0.05). Conclusion The incidence of Ⅲ-Ⅳ degree of aGVHD is unrelated to the amount of grafted MNC, and CD34+cells.

OBJECTIVE: To explore the effection of the pulmonary function of patients of chronic rhinosinusitis (CRS) with asthma which treated with endoscopic sinus surgery (ESS) based comprehensive treatment. METHOD: There were 50 cases of chronic rhinosinusitis with asthma whom met the study criteria. 35 cases enrolled in the tri al group, which treated with endoscopic sinus surgery, and routine perioperative tratment. Another 15 cases as control group which underwent conservative treatment. Both groups underwent the rule treatment of asthma. The main monitoring indexes, which included visual analogue scale (VAS) score, endoscopic Lund-Kennedy score, control of asthma symptoms, the pulmonary function which involved forced expiratory volume in first second (FEV1), forced vital capacity (FVC), the ratio of forced expiratory volume in first second and forced vital capacity (FEV1/FVC) and peak expiratory flow (PEF), were measured in the patients of each groups before surgery, follow-up for 1 year and 3-year. RESULT: Our study found that the VAS score of CRS with asthma was significantly negatively correlated with FEV1 and PEF (P < 0.05), endoscopic Lund-Kennedy score was significantly negatively correlated with PEF (P < 0.05); After the trial group underwent ESS based comprehensive treatment, the improvement of VAS score and endoscopic Lund-Kennedy score of postoperative compared with preoperative and the same period in the control group were significantly (P < 0.05). The difference of the postoperative asthma control rate of trial group after 1 year and after 3 years, respectively, compared with the same period control group were statistically significant (P < 0.05). The preoperative FEV1, FVC, FEV1/FVC and PEF of trial group compared with preoperative were significantly (P < 0.05). Even the difference of them compared with the same period control group were significantly (P < 0.05), except the FVC in the follow-up 3 years (P = 0.088). CONCLUSION The CRS may aggravate asthma symptoms and affect negatively the pulmonary function, and poor asthma control or aggravate may exacerbate the CRS in the course of CRS with asthma patient. With ESS based on combined therapy, it can improve the condition of CRS significantly and improve the control of asthma symptoms and pulmonary function else.
Adolescent ; Adult ; Asthma ; complications ; Chronic Disease ; Endoscopy ; Female ; Humans ; Male ; Middle Aged ; Nose ; surgery ; Pulmonary Ventilation ; physiology ; Rhinitis ; complications ; surgery ; Sinusitis ; complications ; surgery ; Young Adult

OBJECTIVETo set up a feasible method for detection of objective genes from paraffin-embedded tissues of oral leukoplakia (OLK).

METHODSTwenty-five pieces of formalin-fixed, paraffin-embedded tissues of OSCC were selected and ATM gene was detected respectively by 3 methods: the microdissection-nested PCR method, proteinase K-PCR method and conventional phenol-chloroform-PCR method. The positivity rates were compared statistically.

RESULTSThe positivity rates of these 3 methods were 84%, 52% and 64% respectively. Significant difference was found in positivity rate between the microdissection-nested PCR method and the proteinase K-PCR method (P = 0.032).

CONCLUSIONSThe microdissection-nested PCR method merits recommendation because it is more efficient, easy to perform and has the advantage of less sample amount.

Ataxia Telangiectasia Mutated Proteins ; Cell Cycle Proteins ; genetics ; DNA-Binding Proteins ; genetics ; Humans ; Leukoplakia, Oral ; genetics ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Protein-Serine-Threonine Kinases ; genetics ; Tumor Suppressor Proteins ; genetics

Objective To report the experience of lung transplantation with size reduced graft lung.Methods Four cases receiving lung transplantation with size reduced graft lung were analyzed retrospectively.In case 1,left lung transplantation combined with contra-hteral lung volume mduction.In case 2,right lung transplantation Wag individually performed with partially msecfion of upper lobe of graft lung.In case 3.bilateral sequential lung transplantation wag performed using graft lung with partially resection of bilateral upper lobes.In the remained ease,bilateral sequential lung tansplantation was performed using graft lung with resection of right lower lobe.Results All the size reduced graft lungs had good functions during the peri-operation period.Case 1 and case 2 still survived without obvious complication.Case 3 experienced temporary air leak on the 5th day postoperation and cured by water seal drainage but died of abrupt bronchorrhea due to aspergillus infection on the 32th day postoperation.The last cage experienced smoothly recovery excepted fatal virus pneumonia 2 months postopemtion.Conclusion Size reduced graft lungs can be successfully used for transplantation.

OBJECTIVETo investigate individualized one-staged correction of alveolar cleft and lip and nasal deformities secondary to lip cleft.

METHODSThe alveolar cleft and lip and nasal deformities secondary to lip cleft were corrected in one stage.

RESULTSFrom 2004 to 2007, 37 cases were treated. 33 patients were treated successfully with primary healing in bony recipient area. Cancellous bone exposure happened in 3 cases. The wounds healed after debridement and drainage. The cosmetic results were satisfactory.

CONCLUSIONSOne-staged correction of alveolar cleft and the lip and nasal deformities secondary to lip cleft can achieve good results.

Adolescent ; Alveolar Process ; abnormalities ; Child ; Cleft Lip ; complications ; surgery ; Cleft Palate ; complications ; surgery ; Female ; Humans ; Male ; Nose ; abnormalities ; Nose Deformities, Acquired ; surgery

OBJECTIVETo explore a new method of establishing HepG2 cell model of steatosis and observe the expression and significance of nuclear factor erythroid-2p45-related factor 2(Nrf2)/antioxidative response element (ARE) pathway related factors in HepG2 cells of steatosis.

METHODSHepG2 cells were induced with DMEM containing 25% fetal bovine serum, 0.1% MCT/LCT Fat Emulsion and 0.1 mmol/L free fatty acid (FFA) at different stages and the control group cells were cultured with normal DMEM medium. After the cell models were successfully established, lipid droplets in cytoplasm were observed with Oil Red 0 staining, and the triglyceride (TG) accumulation in HepG2 cells were tested by biochemical assay. Intracellular reactive oxygen species (ROS) concentration were detected by flow cytometry. Nitric oxide (NO), superoxide dismutase(SOD), malonyldialdehyde(MDA) and glutathione peroxidase(GSH-Px) were tested by biological reagent kit, while the protein expression of nuclear factor erythroid-2p45-related factor 2(Nrf2), heme oxygenase-1 (HO-1) and

NAD(P)Hquinone oxidoreductase-1(NQO1) were analyzed by Western blot.

RESULTSCompared with that in the control group, red cytoplasmic lipid droplets were visible in model group; TG,ROS, NO, MDA concentration (P < 0.05, P < 0.01) and the protein expression of Nrf2, HO-1 and NQO1 (P < 0.05, P < 0.01)were significantly higher in model group, while SOD, GSH-Px concentration reduced significantly (P < 0.01).

CONCLUSIONThe in vitro cell model of steatosis and oxidative stress was successfully established. The activation of Nrf2/ARE pathway related factors maybe relevant to the overreaction of oxidative stress in HepG2 cells of steatosis.

Antioxidant Response Elements ; Culture Media ; Fatty Acids, Nonesterified ; Fatty Liver ; metabolism ; GA-Binding Protein Transcription Factor ; Glutathione Peroxidase ; metabolism ; Heme Oxygenase-1 ; metabolism ; Hep G2 Cells ; Humans ; Malondialdehyde ; metabolism ; NAD(P)H Dehydrogenase (Quinone) ; metabolism ; NF-E2-Related Factor 2 ; metabolism ; Nitric Oxide ; metabolism ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Triglycerides ; metabolism

To produce specific monoclonal antibody (McAb) against human thrombomodulin (hTM), the full-length hTM cDNA-expressing plasmid pThr402 was transfected into CHO cells by Lipofectamine 2000 reagent. The hTM-expressing CHO cells, which was confirmed by flow cytometry and Western blot, were obtained by G418 selection. Then the McAb against hTM was prepared with classic hybridoma technique. A cell line of CHO-TM5 with high level of hTM was used to immunize female Balb/c mice 3 times at an interval of 4 weeks. On the third day after the third immunization, mice were sacrificed and spleen cells were harvested to prepare hybridoma cells with SP2/0 cells at the ratio of 10 to 1. Hybridoma cells were then cultured at 96-well plates for screening. Cellular enzyme-linked immunoabsorbent assay (CELISA) was applied twice. The first CELISA was done with polythene ELISA plate with a monolayer of CHO-TM5 cells. The positive clones from the first screen were then selected by reacting with similar screening ELISA plate but with CHO cell monolayer instead. Only clones that were positive for the first screening and negative for the second screening were kept, and called as CHO-TM5(+)CHO(-) hybridoma cells. Balb/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded McAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. One line of hybridoma cells with high expression of specific McAb against hTM, NH-1, was obtained. The Ig subclass of the McAb was IgG1 and the titer of ascitic McAb was 1x10(-6). Flow cytometry, CELISA and Western blot assays demonstrated that McAb NH-1 could specifically recognize hTM expressed in CHO-TM5 cells and human umbilical vascular endothelial cells. Meanwhile, the tissue specificity of antigen recognized by McAb NH-1 was identified by immunohistochemical ABC staining. NH-1 can specifically recognize the natural hTM expressed mainly in vascular endothelial cells, which will potentially be useful for investigation of the functions and clinic values of hTM.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; CHO Cells ; Cricetulus ; Female ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Thrombomodulin ; immunology ; Transfection

OBJECTIVETo study whether hematologic malignancy patients with anemia have a lower erythropoietin (EPO) response.

METHODSSerum EPO levels were detected by ELISA in patients with hematologic malignancies and with iron deficiency anemia (IDA). Eighty patients with hematologic malignancies, including 13 multiple myeloma (MM), 7 chronic lymphocytic leukemia (CLL) and 60 non-Hodgkin's lymphoma (NHL) were studied. Thirty of them had anemia(21 NHL,6 MM and 3 CLL). Twenty patients with IDA were studied as the control.

RESULTSHematologic malignancy patients with anemia had higher EPO levels [(97.8 +/- 183.9) IU/L] than those with normal Hb values [(27.8 +/- 85.4) IU/L; P <0.01]. In patients with IDA, serum EPO response was inversely correlated with Hb level (r= -0.5, P <0.05) , but no such inverse correlation was found in the hematologic malignancy patients with anemia (r = -0.14). After corrected for Hb level, the serum EPO levels were significantly lower in anemic patients with hematologic malignancies than in IDA patients (P = 0.032) , indicating a decreased EPO response in the former group.

CONCLUSIONAnemia associated with hematologic malignancy might result from an inappropriately low EPO response. EPO treatment for these patients may be beneficial.

Adolescent ; Adult ; Aged ; Anemia, Iron-Deficiency ; blood ; complications ; Enzyme-Linked Immunosorbent Assay ; Erythropoietin ; blood ; Female ; Hematologic Neoplasms ; blood ; complications ; Hemoglobins ; metabolism ; Humans ; Male ; Middle Aged ; Prospective Studies

OBJECTIVETo investigate the amplification of EMS1 gene in the carcinogenesis of oral mucosa.

METHODSA total of 78 subjects, including 30 patients with oral leukoplakia (OLK), 33 with oral squamous cell carcinoma (OSCC), and 15 healthy controls, were studied. By using microdissection method, we obtained normal mucosa, hyperplastic epithelia, mild-dysplastic epithelia, moderate-dysplastic epithelia, severe-dysplastic epithelia and primary OSCC tissue. Then we analyzed EMS1 amplification by using differential PCR.

RESULTSEMS1 amplification began from moderate-dysplastic epithelia and occurred in 20.0% OLK cases and 57.6% OSCC cases. In the progress of OSCC, no gene amplification was observed in normal tissues, non-dysplastic OLK and mild-dysplastic OLK, while in the cases with metastasis, amplification frequency increased significantly (P = 0.015).

CONCLUSIONSEMS1 amplification parallels with the progress of oral carcinogenesis, indicating their potential roles in oral carcinogenesis.

Carcinoma, Squamous Cell ; genetics ; pathology ; Case-Control Studies ; Cortactin ; genetics ; Gene Amplification ; Humans ; Leukoplakia, Oral ; genetics ; pathology