Objective To explore the therapeutic effect of budesonide combined with salbutamol on serum interleukin (IL-6) and tumor necrosis factorα (TNF-α) in treatment of bronchial pneumonia. Methods A total of 110 cases of bronchial pneumonia patients were randomly divided into the study group and the normal group, with 55 cases in each group, the two groups were given oxygen, anti-inflammatory and symptomatic treatment, the study group received budesonide combined with salbutamol treatment, the curative effect and changes of serum IL-6, TNF-alpha levels were compared between two groups. Results The disappeared time of cough, fever, shortness of breath,rhonchi and moist rales, hospitalization time in study group were significantly shorter than the conventional group, the difference was statistically significant (P<0.05). After 5 days' treatment, the serum IL-6, TNF-α in study group were lower than normal group(P<0.05), the PaO2, SaO2 in study group were higher than normal group (P<0.05). The efficacy in study group was higher than normal group (P<0.05). Conclusion Budesonide combined with salbutamol in treatment of bronchial pneumonia patients could improve clinical symptoms and anti-inflammatory effect, so as to shorten the treatment time and improve the clinical effect.

Objective To investigate the effects of esculin and digitalisglycosides eye drops combined with 532 laser on the treatment of diabetic retinopathy.Methods 65 cases(130 eyes)with diabetic retinopathy in our hospital from November 2012 to November 2015 were selected and divided into observation group(33 cases)and control group(32 cases).The control group was treated with 532 laser treatment,and the observation group was combined with esculin and digitalisglycosides eye drops.Two groups of treatment were one months.Therapeutic effect of two groups were compared,improvement of visual acuity,retinal hemorrhage,exudation,edema absorption time,before and after macular retinal thickness changes.Results After treatment,the total efficiency of the observation group(96.97％)was higher than the control group(75.00％),the difference was statistically significant(P<0.05),the visual acuity of the observation group was significantly higher than that of the control group,and decreased significantly less than the control group,the difference was statistically significant(P<0.05),the retinal hemorrhage,exudation and edema were observed in the observation group faster than the control group,the difference was statistically significant(P<0.05),the macular retinal thickness decreased in the two groups after treatment,the difference was statistically significant(P<0.05),the retinal thickness of the macular area was lower in the observation group than in the control group,the difference was statistically significant(P<0.05).Conclusion Esculin and digitalisglycosides eye drops combined with 532 laser in the treatment of diabetic retinopathy has significant effect,and can reduce the macular retinal thickness,which has important clinical significance.

Objective Getting the robust exogenous gene expression vector under the control of porcine insulin promoter, and to lay the foundation for pancreaticβ-cells specific transgene expressing pigs.Method Using porcine insu-lin promoter ( PIP, 1500 bp of the 5′UTR from the porcine INS gene including the first exon and the first intron) to con-struct expression vector, the HindIII restriction site which connected the sequences of PIP and EGFP was designed before ATG, named PIP-HindIII-EGFP.Considering that the different location of restriction site may affect the expression efficien-cy of the transgene, we optimized the expression vector.Firstly the HindIII restriction site was deleted to realize the seam-less connection of PIP and EGFP,the vector was named PIP-EGFP.Also we mutated the 3′intron splicing acceptor site( SA) of the first intron into HindIII restriction site, named as PIP-SA( M)-EGFP.Three different EGFP expression vectors were respectively transfected MIN-6 mouse pancreatic β-cells, pig ear fibroblasts and kidney cells.The transfected cells were cultured for 48 h and harvested for RT-PCR, flow cytometry and Western blot analysis, to analyze and compare the expres-sion efficiency of vectors.Results After transfection,green fluorescence was observed only in MIN-6 mouse pancreaticβ-cells.RT-PCR analysis and product sequencing showed that the three expression vectors did have different stability with in-tron splicing.The PIP-HindIII-EGFP construct and PIP-EGFP vector produced two kinds of mRNA with the first intron spliced and no spliced, indicating the instability of intron splicing.Mutation of the PIP splice site would cause the first in-tron not spliced, while flow cytometry and Western blot displayed that the mutation induced a most efficient expression of the downstream gene.Conclusions A robust and specific β-cells expression vector has been successfully generated by mutating the intron splicing acceptor site of the porcine insulin promoter.It provides the foundation for preparation of pigs with pancreaticβ-cells specifically expressing the transgene.

Objective To investigate the relationship between the expression of trannsient receptor potential vanilloid 1 (TRPV1) and the severity of airway remodeling in elderly patients with chronic obstructive pulmonary disease(COPD).Methods According to airflow obstruction severity,totally 100 cases of elderly patients with COPD were divided into chronic obstructive pulmonary disease Global Initiative(gold) grade 1 in 23 cases,24 cases of grade GOLD2,GOLD3 27 cases,GOLD4 26 cases,respectively.The TRPV1 concentrations in induced sputum supernatant and serum from each level of elderly patients with COPD as well as in 50 cases of healthy old people were analyzed.Results TRPV1 concentrations in serum and induced sputum in the COPD group was significantly increased compared with the healthy elderly group[(9.94±2.91)μg/L vs.(3.68±0.46)μg/L,(3.29± 1.32)μg/L vs.(0.70 ± 0.30)μg/L] (P ＜ 0.01).The serum and induced sputum TRPV1 concentrations in the mutual pairwise comparison between the elderly COPD patients with all levels had statistical difference (P ＜ 0.01).Conclusion The expression of TRPV1 protein become increased with the severity of airway remodeling in elderly patients with chronic obstructive pulmonary disease.

AIM: To decide the effect that selected siRNA degrades mRNA of IL-1? specifically and suppression of its expression after connected with target site with homology complementary sequence. METHODS: Synthesized DNA expression box aimed directly at target site through PCR reaction in vivo was purified, and transfected into lymphocytes stimulated by LPS. siRNA was transcribed by cellular endogenous RNA polymerase Ⅲ and then evoke the degradation of target mRNA. After 48 hours of transfection, the cell culture supernatant was collected and the concentration of IL-1? was assayed using ELISA. RESULTS: Compared with blank-control and negative-control, selected sequence decreased the expression of IL-1?. Rate of the suppression was about 15%. CONCLUSION: RNAi technology produces specific interference effect in mouse spleen lymphocytes in original culture and inhibits the excretion of IL-1?.

ObjectiveTo assess the differences of short- and long-term postintervention status on ocular and systemic symptoms for patients with ocular myasthenia gravis (OMG) after pyridostigmine bromide, corticosteroid, thymectomy, or thymectomy-corticosteroid combination therapy ( combination ).MethodsThis retrospective plus prospective study included 180 OMG patients, whose age of onset ≥ 15 years, treated non-randomly with above therapies separately: thymectomy group (60 cases ), corticosteroid group (39 cases), combination group ( 31 cases ), symptomatic group ( 50 cases ). Postintervention status complying with Myasthenia Gravis Foundation of America (MGFA)complete stable remission ,pharmacologic remission, or minimal manifestations was considered as desirable response, which was used as statistical indicator. Results ①Corticosteroid group showed higher desirable response rates on ptosis, ophthalmoplegia and general weakness at 3-6 months after treatment than other groups, and 42. 1％( 16/38 ) of them at 3 months achieved the desired state of ptosis, higher than the symptomatic group (7/48,14. 6％, ×2 = 8. 200, P = 0. 004 ). ② Ascending ideal rates had been presented in both combination and thymectomy groups since 1 year after treatments, while a little bit higher rate was presented in the former. At the end of observation, 21.7％ ( 13/60 )of patients in thymectomy group achieved complete stable remission.By paired longitudinal comparisons,thymectomy group showed higher ideal rates on ptosis (22/40,55.0％ ), ophthalmoplegia ( 16/27,59. 3％ ) and general weakness (20/40,50. 0％ ) at 2 years than that at 3 months( 11/59,18.6％ ;11/44,25.0％ ;9/60,15.0％ ;P =0. 002, 0. 031,0.000). ③For those patients by symptomatic treatment, the average age of onset was (51.9 ± 18.0) years, higher than that by other 3 therapies (F = 10. 563 ,P =0. 000). ④OMG patients with ophthalmoplegia more likely select corticosteroid or combined therapy. Ophthalmoplegia in combination group was higher than that in symptomatic and surgery groups( ×2 = 12. 939,14. 380, P =0. 000 in both). Ophthalmoplegia in corticosteroid group was higher than that in surgery group ( ×2 = 8. 017, P = 0. 005 ).Conclusions Corticosteroid appears to early overcome ptosis, ocular motor dysfunction and general weakness for patient with OMG in early-to-middle adulthood.Thymectomy andsurgery-corticosteroid combinationtherapies bothshowlong-term effectonthem.

[Objective]To establish a novel noninvasive fluorescent animal model for endometriosis in vitro and in vivo.[Methods]Adenovirus encoding enhancing green fluorescent protein(Ad-eGFP)was used to transfect endometrial glandular cells and stromal cells(cells transfection and injection,Method No.1),and fragments(tissues transfection and injection,Method No.2).Transfection efficiencies were compared between the two methods in vitro.Then GFP transfected glandular cells and stromal cells suspension were injected into nude mice subcutaneously(Method No.1),taking Method No.2 as a comparison.In vivo observation last for 25 days,and positive rates and duration times of fluorescent lesions were calculated.Histological examination was used to confirmed lesion formation.[Results]On the fifth day after injection,lesion positive rate of Method No.1 was 88.9%,which was statistically significantly higher than that of Method No.2(22.2%),P=0.015＜0.05.The fluorescent positive duration of Method No.1 and No.2 were 12 ± 8 days and 7±4 days.The structures of lesions were all identified as human original endometrium by histological examination,including HE staining and immunofluoresceney.[Conclusion]Noninvasive animal model of endometriosis can be built up by subcutaneously injection of Ad-EGFP transfected endometrial glandular cells and stromal cells suspension with higher positive rate and longer observation time

Objective To investigate the feasibility of combined laparoscopic and thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity.Methods We retrospectively analyzed the clinical data of 38 patients who underwent esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity from October 2011 to August 2012.To remove the stomach in laparoscopic and the esophagus in thoracoscopy.The main portion of a gastric conduit is created using three to four firings of a linear stapler(Ethicon Endo-surgery,Cincinati,OH) and jejunum stoma.Gastric conduit was pulled into the chest cavity and anastomosed to the esophagus.Results The average operative time was 280 minutes,the mean operative blood loss was 120 ml.No patient required laparotomy.No pulmonary complications or anastomotic leaks occurred.One had gastric retention,another one had chylous hydrothorax.All patients were cured,no one dead in hospital.Conclusion Combined laparoscopic and thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity is technically feasible and safe,minimized trauma,less operative blood loss and quick recovery.

Objective To explore thc clinical manifestation of secondary generalized myasthenia gravis(GMG) and analyze the factors affecting the progression from ocular myasthenia gravis(OMG) to GMG.Methods This research constitutes a single-center,retrospectively-collected prospective cohort study.We comprehensively reviewed our self-managed myasthenia gravis (MG) database drawn from personal clinical experience from January 2000 to Junc 2013.Patients underwent series of examination including repetitive nerve stimulation (RNS) tests,measurement of serum acetylcholine receptors antibody and serum muscle-specific tyrosine kinase antibodies,thymus computer tomography scan etc.Patients were treated with pyridostigmine bromide,corticosteroid therapy and (or) thymectomy based on a nonrandomization pattern and they were documented for their respective symptoms of OMG and GMG and date of GMG conversion.Logistic regression analysis was adopted to determine the influencing factors correlated with the development of GMG during the follow-up.Results Totally 770 patients initially diagnosed with OMG were included,among whom 573 (74％) patients remained with OMG (R-OMG group) and 197(26％) patients developed into GMG (GMG group) during the follow-up.(1) In comparison with their R-OMG counterparts,patients with secondary GMG were older at onset; Displayed more frequent RNS abnormality of facial nerve,accessory nerve and ulnar nerve ; Showed higher incidence of thymoma and were less treated by early corticosteroids.(2) For GMG group,81％ (160/197) of them displayed bulbar MG; 67％ (132/197) of GMG conversion occurred within 2 years,and 84％ (166/197) within 5 years.In comparison with the patients with onset of≤ 14-year-old,both of patients with15-49-year-old and≥ 50-year-old displayed higher conversion rate and shorter conversion duration (median:10 years versus 1 year and 6.5 months).(3) RNS abnormality of accessory nerve(OR =6.650,95％ CI 3.547-12.471 ; P ＜ 0.05) and thymoma(OR =7.924,95％ CI 2.554-24.585 ; P ＜ 0.05) were prognostic factors for the development of GMG,while early corticosteroid(OR =0.232,95％ CI 0.119-0.452 ; P ＜ 0.05) predicted the reduction of the risk of generalization.Conclusions Multiple factors including abnormal RNS of proximal limb muscles,thymoma,early corticosteroids therapy and possibly even onset age of over 15-year-old may involve the generalization in patients with OMG at onset.

Objective To identify the rate of hepatitis B virus (HBV) reactivation and potential risk factors in hepatitis B surface antigen negative/hepatitis B core antibody positive patients with lung cancer receiving adjuvant chemotherapy without concomitant antiviral prophylaxis.Methods The records of 3280 patients with lung cancer who received adjuvant chemotherapy were retrospectively reviewed from January 2003 to December 2011.Among these patients,367 hepatitis B surface antigen negative/hepatitis B core antibody positive patients were analyzed for the HBV reactivation in this study.The HBV serology marker and biochemical tests of the 367 patients were performed.The data were analyzed by chi square test.Results Among 367 hepatitis B surface antigen negative/hepatitis B core antibody positive patients with lung cancer,14 patients suffered HBV reactivation.Univariate analysis showed that age≥70 years(x2 =13.003,P=0.019),abnormal liver computed tomography findings (x2 =11.225,P =0.026) and the amount of corticost eroids≥ 150 mg(x2 =7.008,P =0.033)were associated with HBV reactivation.However,gender and adjuvant chemotherapy regimens were not related with HBV reactivation.Conclusion HBV reactivation occurs in a proportion of hepatitis B surface antigen negative/hepatitis B core antibody positive patients with lung cancer during adjuvant chemotherapy.