Aim To study the role of TNF-α/NF-κB signaling in matrix metalloproteinase ( MMP)-9 expres-sion induced by lipopolysaccharide ( LPS ) in airway epithelial cells, and to investigate the effects of lenti-virus mediated RNAi targeting a disintegrin and metal-loproteinase 17 ( ADAM17 ) gene on MMP-9 expression induced by LPS. Methods The ADAM17 siRNA ex-pression vector was constructed, and packaged to re-combinant lentivirus in 293T cells. The HBE4-E6/E7 cells were pretreated for 30 min by NF-κB inhibitor ( PDTC) and a recombinant human TNFR p75-Fc fu-sion protein ( Etanercept) , or infected by the recombi-nant lentivirus for 72 h, and then stimulated for 24 h by LPS or TNF-α. The release of TNF-α was detected by ELISA. The mRNA and protein levels of MMP-9 were analyzed respectively by RT-PCR and Western blot. NF-κB activity was detected by electrophoretic mobility shift assay. Results LPS and TNF-α signifi-cantly increased MMP-9 mRNA and protein expressions and the activation of NF-κB in HBE4-E6/E7 cells ( P<0. 05 ) . Etanercept and PDTC significantly inhibited MMP-9 expression and the activation of NF-κB induced by LPS ( P<0. 05 ) . Lentivirus mediated RNAi targe-ting ADAM17 significantly decreased TNF-α produc-tion, inhibited MMP-9 mRNA and protein expressions and the activation of NF-κB induced by LPS in HBE4-E6/E7 cells ( P <0. 05 ) . Lentivirus mediated RNAi targeting ADAM17 did not inhibit MMP-9 mRNA and protein expressions and the activation of NF-κB in-duced by TNF-α ( P>0. 05 ) . And PDTC significantly inhibited MMP-9 mRNA and protein expressions and the activation of NF-κB induced by TNF-α ( P <0. 05 ) . Conclusions TNF-α/NF-κB signaling partic-ipates in the regulation of MMP-9 expression induced by LPS in airway epithelial cells, and lentivirus-media-ted RNAi targeting ADAM17 plays an important role in that signaling pathway upstream by regulating TNF-αrelease.

Objective To explore the outcome of modified bladder neck reconstruction in treating the urinary incontinence due to the sphincter dysfunction.Methods Between January 2010 and December 2014,a total of 23 patients,including 16 male and 7 female cases,with incontinence due to sphincter dysfunction had undergone a procedure of modified bladder neck reconstruction.The mean age was 36 years (range 17-61 years).Etiology of incontinence was secondary to pelvic fracture and urethral rupture procedure in 19 patients and other failure procedures in 4 cases.The mean duration of incontinence was 2 years (range 1-5 years).Urodynamic examination was performed in all patients and the mean maximum urethral pressure was 34 cmH2O (range 21-43 cmH2O).The modified bladder neck reconstruction was performed in 23 patients.Under the general anesthesia and vertical bladder incision,triangular mucosal strips,from the bladder neck to ureteral office,were denuded and leaving the central urethral plate,which was 2.0-2.5 cm in width.In order to form a neourethra and bladder neck,the multilayer triangular muscles were sutured interruptedly over a 14-16 F catheter using 2-0 or 3-0 polyglactin.And the anterior wall of the bladder was then closed over the new bladder neck using continuous sutured.Results The 23 patients were followed up for 6 months to 3 years,mean 1.4 years.Continence achieved in 5 patients.Of them,2 patients had difficulty in voiding but corrected by indwelling the catheter for another 2 weeks.Social continence was achieved in 7 patients.Incontinence status was improved in 7 cases and failed in 2 cases.Urodynamic examination was performed in 4 patients and the mean maximum urethral pressure was 64 cmH2O (range 52-72 cmH2O).Conclusions Our study suggested that the modified bladder neck reconstruction was a good procedure for the treatment of urinary incontinence due to sphincter dysfunction,particularly for the incontinence secondary to the traumatic urethral stricture or other operation.

OBJECTIVETo construct certain chimeric E3s expression plasmids targetting oncoprotein Ras by harnessing the theory of protein knockdown.

METHODSWe chose the binding domain of Raf-1, PI3K, RalGDS, and the function domain of F-Box as well as the U-Box to construct the plasmids. Then used the double enzyme, PCR, and sequence to test the validity and integrity of the cloned nucleotide fragments. The expression efficiency of the plasmids in eukaryotic cells was detected by Western blot analysis.

RESULTSFive of 6 plasmids in this study expressed the corresponding fusion proteins in HEK293T cells, and (RBD+CRD)(Raf-1)- U-Box-pcDNA3.1 can knocked down the protein level of Ras in PANC-1 cells.

CONCLUSIONSWe successfully constructed the chimeric E3 expression plasmids, which provides a solid basis for further research on protein knockdown.

Cloning, Molecular ; Genetic Vectors ; HEK293 Cells ; Humans ; Phosphatidylinositol 3-Kinases ; genetics ; Plasmids ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Ubiquitin-Protein Ligases ; genetics ; ral Guanine Nucleotide Exchange Factor ; genetics ; ras Proteins ; genetics

Ventricular remodeling is one of the main causes of heart failure. A large number of studies have shown that inflammation plays an important role in the occurrence and development of ventricular remodeling. Recent studies found that chronic inflammation mediated by T cells is closely related to the progression of ventricular remodeling. This review summarized the recent research progress of T lymphocyte subsets in ventricular remodeling.

OBJECTIVETo investigate long-term survival after multivisceral resection in patients with locally advanced right colon cancer.

METHODSThe clinical data and survival of 13 patients with locally advanced right colon cancer were retrospectively analyzed.

RESULTSThere were 8 males and 5 females with a mean age of 58.6 years. Location of the primary tumor included hepatic flexure(n=6), transverse colon(n=2), and ascending colon(n=5). Three patients had duodenal invasion alone, 9 had involvement of duodenum and other organs, and 1 had pancreas and stomach involvement. Right colectomy and pancreaticoduodenectomy and(or) resection of other organs were performed. The 1-, 3-, and 5-year survival rates were 69%, 54%, and 30%, respectively.

CONCLUSIONRight colectomy combined with multivisceral resection is a promising procedure for selected patients with locally advanced colon cancer.

Adult ; Aged ; Colonic Neoplasms ; surgery ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.

OBJECTIVE: To investigate the prevalence rate, malocclusion type and treatment rate as well as awareness of malocclusion among children and adolescents living in the Hangzhou municipality. METHODS: 1818 children and adolescents ages 7 approximate, equals 16 living in Hangzhou municipality were evaluated. RESULTS: The overall malocclusion rate was 35.75%. In Angle's classification: Angle I 593 cases (32.62%), Angle II 48 cases(2.64%),Angle III 9 cases (0.49%). Among the malocclusion type of crowding was mixed dentition 70.66%,permanent dentition 85.89%. In the overjet malocclusion mixed dentition was noted in 60.57% and permanent dentition in 51.05%.The type of overbite was mixed dentition 67.82%,permanent dentition 31.23%. In the crossbite of anterior teeth mixed dention was noted in 12.30% and permanent dention in 9.91%. Overall treatment rate for malocclusion was 10.15%.CONCLUSION: Among Hangzhou municipality juveniles there is both inadequate prevention and treatment of dental malocclusion.

BACKGROUNDFanconi anemia is a severe congenital disorder associated with mutations in a cluster of genes responsible for DNA repair. Arriving at an accurate and timely diagnosis can be difficult in cases of Fanconi anemia with atypical clinical features. It is very important to increase the rate of accurate diagnosis for such cases in a clinical setting. The purpose of this study is to explore the clinical diagnosis of Fanconi anemia in children with atypical clinical features.

METHODSSix cases of Fanconi anemia with atypical clinical features were enrolled in the study, and their clinical features were recorded, their FANCA gene transcription was assessed by RT-PCR, and FANCA mutations and the ubiquitination of FANCD2 protein were analyzed using DNA sequencing and western blotting respectively.

RESULTSAll six cases showed atypical clinical features including no apparent deformities, lack of response to immune therapy, and progressively increasing bone marrow failure. They also have significantly increased fetal hemoglobin, negative mitomycin-induced fracture test results, and carry a FANCA gene missense mutation. Single protein ubiquitination of FANCD2 was not observed in those patients.

CONCLUSIONThe combination of clinical features, FANCA pathogenic gene mutation genotype and the absence of FANCD2 protein ubiquitination are helpful in the accurate and timely diagnosis of Fanconi anemia in children.

Child ; Child, Preschool ; Fanconi Anemia ; diagnosis ; genetics ; metabolism ; Fanconi Anemia Complementation Group D2 Protein ; genetics ; metabolism ; Female ; Humans ; Male ; Mutation ; Ubiquitination

OBJECTIVETo investigate the anatomic characteristics of splenic flexure, surgical techniques, and oncologic outcomes in 52 patients with non-obstructive splenic flexure colon cancer.

METHODSClinical data of 52 patients with non-obstructive splenic flexure colon cancer from March 2004 to March 2011 in the Department of General Surgery at the Henan Province Tumor Hospital were analyzed retrospectively.

RESULTSThere were 37 patients of regular type, 5 of mobile type, and 10 of adhesive type. All the patients received radical operation. Eighteen patients received pre-small intestine anastomosis, including 12 cases with regular type, 4 with mobile type, and 2 with adhesive type. The difference in pre-small intestine anastomosis among the three types was not statistically significant(P=0.062). In addition, 32 cases received retro-ileum anastomosis. There were no significant differences in operative time, intraoperative blood loss, number of lymph node dissection and positive lymph node, and postoperation complication rate among the three types. Follow up was available in all the cases. Five-year survival rates of cases with regular type, mobile type and adhesive type were 62.5%, 59.2% and 58.7% respectively(P>0.05).

CONCLUSIONSRadical resection can provide satisfactory survival for splenic flexure colon cancer patients. The anatomy of splenic flexure does not affect the type of anastomosis. Retro-ileum anastomosis is a simple and effective method for reconstruction after radical resection of the tumor.

Adult ; Aged ; Anastomosis, Surgical ; Colon, Transverse ; anatomy & histology ; pathology ; surgery ; Colonic Neoplasms ; pathology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the effect of compound qingqin liquid (CQL) on Toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) in rats with urate nephropathy, and to explore its renal protection mechanism.

METHODSTotally 55 SD rats were randomly divided into 5 groups, i.e., the normal control group (n =5), the model group (n =10), the positive drug group (n=10), and the high-, medium-, low-dose CQL groups (n=10) respectively. The urate nephropathy model was induced by intragastrically administering adenine and feeding yeast. Distilled water was intragastrically administered at the daily dose of 10 mL/kg to rats in the normal control group and the model group. Allopurinol was intragastrically administered at the daily dose of 9.33 mg/kg to rats in the positive control group. CQL was intragastrically administered at the daily dose of 3.77, 1.89, 0.94 g/kg to rats in the high-, medium-, and low-dose CQL groups. Rats of each group were executed in batches at the 4th and 6th week respectively. Their kidney tissues were taken out to determine the mRNA transcription level of TLR2 and TLR4 by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression level of TLR2 and TLR4 were determined by Western blot. The protein expression level of TLR4 was also detected by immunohistochemical assay.

RESULTSAt week 4 and 6, the protein expression of TLR2 and TLR4 as well as the mRNA transcription of TLR4 increased in the model group, when compared with the control group (P < 0.05, P < 0.01). Compared with the model group, there was no statistical difference in the transcription level of TLR2 mRNA or TLR4 mRNA among the 3 CQL groups (P > 0.05) at week 4 and 6. Additionally, at week 6, the protein expression of TLR4 and TLR2 could be reduced by CQL (P < 0.05, P < 0.01).

CONCLUSIONCQL might protect kidney tissue against inflammatory injury by inhibiting the protein expression levels of TLR2 and TLR4.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; drug effects ; metabolism ; Kidney Diseases ; drug therapy ; metabolism ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Uric Acid