To study the relationship between asthma and bronchial hyper responsiveness(BHR).The reactivity of bronchi to inhalation of methacholine, the plasma levels of substance P(SP),soluble P selection (sP selectin),vasoactive intestinal peptide(VIP), and fibronectin(Fn) were assessed in 68 cases of asthma, 62 patients with cough variant asthma(CAV), and 43 healthy subjects.It was showed that the tests of airway reactivity were all positive in 68 cases of asthma and 62 cases of CVA.Dmin in asthma and CVA was(8 81?9 67)u and(5 97?4 37)u,respectively,in these two groups of patients. Dmin in asthma patients was higher than in CAV patients ( P

Adult ; Carcinoma, Hepatocellular ; secondary ; Humans ; Liver Neoplasms ; secondary ; Lymphoma, Non-Hodgkin ; pathology ; Male ; Spinal Neoplasms ; pathology

In recent years,laparoscopic total gastrectomy (LTG) with lymphadenectomy is increasingly utilized for the management of gastric cancer located in the middle or upper third of the stomach.However,esophagojejunostomy is the key technical difficulty in operation.Compared with conventional extracorporeal esophagojejunostomy via mini-laparotomy,pioneers are attempting to perform intracorporeal anastomosis in order to gain better manipulation and minimally invasive benefits,as well as reducing the difficulties in digestive tract reconstruction.

Objective To investigate the effects of silencing Smo gene on proliferation and apoptosis of rat prima-ry chondrocyte in vitro.Methods The primary chondrocyte was obtained by mechanical-enzyme digestion and identified by Immunohistochemical cells ( ColⅡ) .The animals were divided into control group , control siRNA group and Smo siRNA 1 ~3 group.The siRNA was transfected into chondrocytes by lentivirus vector .After 72 h, the cell viability was detected by MTT, Smo expression was detected by RT-PCR and Western blot, and the apoptosis of chondrocyte was assessed by flow cytometry .Results All types of siRNA were transfected into primary chondrocyte by vectors, the Smo siRNA 1 ~3 may inhibit the expression of Smo mRNA and protein in chondrocytes, and Smo siRNA2 had the highest silencing rate ( the expressions of Smo mRNA and protein were 0.19 ±0.03 and 0.39 ±0.07 ) .The cell viability in Smo siRNA2 group was lowest ( 77.38% ±7.19%) , while the apoptosis rate of Smo siRNA2 was highest ( 21.43%±2.97%) .Conclusions Silencing Smo gene in primary chondrocytes may inhibit proliferation and promote apoptosis , Smo may have a protecting role from apop-tosis of the chondrocyte.

Objective To study the proliferation and apoptosis and investigate the expression of Indian hedgehog (Ihh),parathyroid hormone-related peptide (PTHrp),smoothened (Smo) protein and mRNA in the cultured rat primary chondrocytes exposed to different doses of NaF.Methods The third generation articular chondrocytes of neonate rat were cultured in vitro and treated with 0 (control),5,10,20 and 40 mg/L of fluoride.The proliferation activities of cells at different times (24,48 and 72 h) were tested by Thiazolyl Blue Tetrazolium Bromide (MTT).The apoptosis rate was determined by flow cytometry.The expressions of protein and mRNA of Ihh,Smo and PTHrp at 48 h were determined by Western blotting and semi-quantitative RT-PCR,respectively.Results After exposed to 5 mg/L of fluoride for 24,48 and 72 h,the proliferation rates were significantly increased [(1.17 ± 0.07)％,(1.20 ±0.06)％,(1.16 ± 0.08)％] compared with those of control group [(1.10 ± 0.08)％,(1.13 ± 0.08)％,(1.15 ± 0.08)％],but the proliferation activity at 48 and 72 h in 40 mg/L group [(0.72 ± 0.11)％,(0.68 ± 0.04)％] was significantly lower than those in control group (all P ＜ 0.05).Compared with the control group,apoptosis rate of cartilage cell in fluoride treatment group increased gradually [(1.47 ± 0.05)％,(19.87 ± 3.03)％,(25.30 ± 1.28)％,(45.73 ± 4.63)％,F =123.328,P ＜ 0.01].Western blot analysis and RT-PCR results showed that the Ihh,PTHrp,Smo mRNA and protein expression increased in the fluoride groups at 48 h (Ihh protein:0.77 ± 0.08 vs.0.98 ±-0.07,1.23 ± 0.06,1.37 ±0.07,1.34 ± 0.07;PTHrp protein:0.68 ± 0.04 vs.0.89 ± 0.05,0.83 ± 0.05,1.29 ± 0.05,1.16 ± 0.08;Smo protein:0.37 ± 0.01 vs.0.64 ± 0.06,0.67 ± 0.03,0.96 ± 0.06,0.69 ± 0.06;Ihh mRNA:0.77 ± 0.05 vs.0.98 ± 0.05,1.09 ±0.05,1.27 ± 0.03,1.46 ± 0.06;PTHrp mRNA:0.67 ± 0.07 vs.0.97 ± 0.05,1.07 ± 0.08,1.37 ± 0.05,1.45 ± 0.05;Smo mRNA:0.45 ± 0.03 vs.0.63 ±-0.04,0.71 ± 0.05,0.81 ± 0.01,1.00 ± 0.02,all P ＜ 0.05).Conclusions Low doses of fluoride can promote the proliferation of chondrocytes cultured in vitro,and high doses of fluoride can promote the apoptosis of chondrocytes cultured in vitro.The expression of Ihh signaling pathway RNAs and proteins of the cartilage cells are increased following increased levels of fluoride.The results suggest that fluorine has activated the Ihh signaling pathway in chondrocytes and promoted the proliferation and apoptosis processes which might be involved in chondrocytes injury.

In order to investigate the inhibitory effects of Endostar (rh-endostatin, YH-16) in combination with radiotherapy on lung adenocarcinoma A549 in mice and the interaction mechanisms of combined therapy, the transplantation tumor models of A549 lung adenocarcinoma were established. When the largest diameter of tumor reached 1.0 cm, all nude mice were randomly divided into 4 groups: Endostar group, radiotherapy group, radiotherapy plus Endostar (combined treatment) group, and control group (n=6 in each group). The largest diameter and the vertical diameter of tumor were measured at different time points. At the 16th day, mice were executed, and the tumors were applied to analysis of rate of tumor cell apoptosis, and the expression levels of basic fibroblast growth factor (bFGF) mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR) and those of vascular endothelial growth factor (VEGF) by immunohistochemistry. The results demonstrated that the rate of tumor inhibition in combined treatment group was higher than that in other groups. And the rate of tumor cell apoptosis in combined treatment group was also higher than that in other groups. Meanwhile, the levels of bFGF mRNA and VEGF expression in combined treatment group were lower than those in other groups. It was concluded that Endostar obviously enhanced the curative effectiveness of radiotherapy on lung adenocarcinoma A549 in mice. The underlying mechanisms may involve the down-regulation of bFGF mRNA and VEGF expression to inhibit angiogenesis by Endostar and the cooperative effect of Endostar and radiotherapy to synergistically promote tumor cell apoptosis. And Endostar inhibits angiogenesis by down-regulating the expression of bFGF mRNA and VEGF.

Objective To investigate the levels of serum soluble CDI4(sCD14)in patients early after renal transplantation and the relationship between sCD14 and allograft rejection.Methods This se- ries included 51 consecutive patients undergoing renal transplantation.We prospectively determined levels of serum sCD14 at 1 h before transplantation(d 0)and the first 10 d after transplantation(d 1-10).The ser- um creatinine(Cr)levels were recorded at the same time.The patients were divided into 2 groups(rejection and non-rejection groups)according to whether they experienced acute rejection or not during the first 14 d after transplantation.The levels of serum sCD14 and Cr between the 2 groups were compared.Results Of 51 cases,13 experienced acute rejection,and the mean time from postoperation to rejection start was 7 d;38 cases had no rejection.On d 0,the Cr levels of rejection group[(789?221)?mol/L]and non-rejection group[(742?234)?mol/L]had no significant difference(P＞0.05).The Cr level was higher in rejection group than in non-rejection group on d 1-10.In the 2 groups,the Cr levels of d 3 and d 5 to d 10 were (237?104)vs(160?70),(176?85)vs(117?46),(174?81)vs(112?40),(173?81)vs(112?39),(209?53)vs(112?38),(203?73)vs(103?35),(181?50)vs(102?31)?mol/L,respective- ly,with significant difference between them(P＜0.05).The serum sCD14 levels on d 0 in rejection group [(9.55?5.71)mg/L]and non-rejection group[(8.99?3.89)mg/L]had no significant difference.The sCD14 levels were higher in rejection group than in non-rejection group on d 1-5[(15.52?6.60)vs (9.85?4.11),(15.48?5.85)vs(7.53?3.79),(12.15?4.45)vs(5.88?3.95),(10.84?4.11) vs(4.88?3.17),(7.61?5.37)vs(4.66?1.91)mg/L,respectively]with significant difference(P＜0.05).The sCD14 levels in the 2 groups on d 1 were elevated compared with those on d 0,then decreased gradually.Conclusions It is suggest that the increase in serum sCD14 levels occurs earlier than clinically acute rejection.The serum sCD14 levels on d 1-5 after transplantation can serve as important predictors for acute renal graft rejection.

It is often necessary to construct more than one recombinant plasmids when investigating the characteristics, physchemical features and functional mechanisms of genes or proteins. Repeated sub-cloning procedures including design of primers, enzyme digestion, ligation and verification of recombinant plasmids, have to be involved with. For this reason, it has become a tendency to developing new genetic vectors which can be used in multitude of experiments. Therefore, by using pIRES vector as a backbone, here we reported the construction of a mammalian expression vector: pCMV-Myc-IRES-EGFP which contains the N-terminal c-Myc epitope tag and the enhanced green fluorescent protein (EGFP) translated in an IRES-dependent manner. This novel vector can be used to testify the efficiency of cell transfection, to collect successfully transfected cell population via cytometry, to conduct transcription and translation in vitro, to purify target proteins or to trap their interactional proteins. The availability of this vector can facilitate function study of genes.
Apoptosis Regulatory Proteins ; genetics ; metabolism ; Base Sequence ; Blotting, Western ; Cell Line ; Cloning, Molecular ; Gene Expression ; Genes, myc ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Molecular Sequence Data ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

OBJECTIVETo observe the clinical effect of Huikangling Tablet (HT, extracted from Scabrous Patrinia root) on peripheral blood micrometastasis of differentiated thyroid carcinoma (DTC) patients.

METHODSTotally 87 DTC patients with positive micrometastasis were randomly assigned to the treatment group (45 cases) and the control group (42 cases). DTC endocrine inhibition treatment standards were executed in all patients. They all took levothyroxine sodium (50 microg/tablet, from low dose, 25 microg each time, once per day, 0.5 h before breakfast), and its dosage was gradually added one week later. The dosage was adjusted according to tested results of TSH combined recurrence risk stratification and endocrine suppression induced adverse reactions risk stratification. Patients in the treatment group took HT (0.4 g per tablet, 3 tablets each time, three times per day for a total of 12 weeks) combined TSH suppression therapy, while those in the control group only received TSH suppression therapy. Peripheral micrometastatic cytokeratin 19 (CK19) and polymorphic epithelial mucin1 (MUC1) were detected by FCM at week 4 and 12. Meanwhile, distant metastasis and adverse reactions were observed.

RESULTSAfter 4-week treatment positive micrometastasis was shown in 18 cases (40%) of the treatment group and 29 cases (69%) in the control group with statistical difference (chi2 = 5.68, P < 0.05). After 12-week treatment positive micrometastasis was shown in 7 cases (15.6%) of the treatment group and 17 cases (44.7%) in the control group with statistical difference (chi2 = 8.49, P < 0.01). Pulmonary metastasis occurred in 2 cases and bone metastasis in 1 case of the control group at follow-ups. Cervical lymph node metastasis without accompanied recurrence of thyroid cancer occurred in one case of the treatment group. No obvious liver or renal abnormalities occurred.

CONCLUSIONHT inhibited peripheral blood micrometastasis of DTC patients and its mechanism needed to be further studied.

Antineoplastic Agents ; pharmacology ; therapeutic use ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Neoplasm Micrometastasis ; drug therapy ; Neoplasm Recurrence, Local ; Tablets ; Thyroid Neoplasms ; drug therapy

Brown-Sequard Syndrome ; etiology ; Cervical Vertebrae ; Humans ; Intervertebral Disc Displacement ; complications ; surgery ; Male ; Middle Aged