Adolescent ; Adult ; Aged ; Child ; Endoscopes ; Female ; Humans ; Male ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; Smell ; Turbinates ; surgery ; Young Adult

In gene manipulation, different selectable markers with various linkers are necessary. In order to get selectable markers directly, we constructed from pBlueScript SK(-) a series of particular plasmids, pSKsymKm, pSKsymBle, pSKsymEry, pSKsymHyg and pSKsymGm, each contains Kanamycin, Bleomycin, Erythromycin, Hygromycin or Gentamycin resistance cassette. By restriction enzyme digestion and gel extraction, any of five antibiotic resistance genes with specific ends can be conveniently obtained.

Objective To obtain the gene fragment encoding human mannan-binding lectin (MBL)-associated serine protease-2 (MASP-2). Methods The target cDNA fragment was amplified from the total RNA extracted from human fetal liver tissue by RT-nested PCR, cloned into pGEM-T vector and identified by restriction analysis and sequencing. Results The cDNA fragment encoding MASP-2 polypeptide with the signal sequence was isolated, linked with pGEM-T vector and transformed into E.coli TG1. The restriction maps of the selected clones were consistent with those generated by computer analyses. The result of sequencing of one selected clone showed that its sequence was identical with that of reported human MASP-2 cDNA. Conclusion The human MASP-2 gene was successfully cloned, which prepares the ground for studying the molecular mecha- nisms by which MBL activates the complement lectin pathway and the mutations of MBL gene cause immunodeficiency.

Objective To obtain the gene fragment encoding human mannan-binding lectin (MBL)-associated serine protease-2 (MASP-2). Methods The target cDNA fragment was amplified from the total RNA extracted from human fetal liver tissue by RT-nested PCR, cloned into pGEM-T vector and identified by restriction analysis and sequencing. Results The cDNA fragment encoding MASP-2 polypeptide with the signal sequence was isolated, linked with pGEM-T vector and transformed into E.coli TG1. The restriction maps of the selected clones were consistent with those generated by computer analyses. The result of sequencing of one selected clone showed that its sequence was identical with that of reported human MASP-2 cDNA. Conclusion The human MASP-2 gene was successfully cloned, which prepares the ground for studying the molecular mecha- nisms by which MBL activates the complement lectin pathway and the mutations of MBL gene cause immunodeficiency.

Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Carbon Disulfide ; adverse effects ; antagonists & inhibitors ; Cells, Cultured ; Endothelium, Vascular ; drug effects ; Humans ; Umbilical Veins ; cytology ; drug effects ; metabolism

OBJECTIVETo explore the efficacy and feasibility of dog days moxibustion plaster therapy in treatment of allergic rhinitis of different patterns/syndromes.

METHODSAllergic rhinitis of lung deficiency and invasion of cold, spleen qi deficiency and kidney yang deficiency, 56 patients for each pattern/syndrome were randomized into a plaster therapy group and a nasal spray group, 28 cases in each one. In the plaster therapy group, according to the pattern/syndrome differentiation, with literature retrieval method, 3 acupoints of high frequency utility in clinic were selected as one group in acupoint plaster therapy. For lung deficiency and invasion of cold pattern/syndrome, Feishu (BL 13), Fengmen (BL 12) and Hegu (LI 4) were selected. For spleen qi deficiency pattern/syndrome, Pishu (BL 21), Zusanli (ST 36) and Dazhui (GV 14) were selected. For kidney yang deficiency pattern/ syndrome, Shenshu (BL 23), Dingchuan (EX-B 1) and Bailao (EX-HN 15) were selected. Separately, on July 13, 2013, July 23, 2013, August 2, 2013 and August 12, 2013, the aucpoint plaster therapy was applied, 2 to 4 h (1 to 2 h for children) each time. In the nasal spray group, beclometasone dipropionate aqueous nasal spray, 2 presses one nostril each time, 2 to 3 times a day, continuously for 4 weeks. The symptom score and efficacy were compared before and after treatment in the patients of the two groups.

RESULTSThe symptom scores of 3 patterns/syndromes were all apparently improved after treatment as compared with those before treatment in the patients of the two groups (all P<0.05), and the result in the plaster therapy group was better than that of the nasal spray group (P<0.05, P<0.01). For lung deficiency and invasion of cold pattern/syndrome, the total effective rate was 87.3% (20/24) in the plaster therapy group, better than 84.6% (22/26) in the nasal spray group (P<0.05). For spleen qi deficiency pattern/syndrome, the total effective rate was 83.3% (20/24) in the plaster therapy group, obviously better than 76.9% (22/26) in the nasal spray group (P<0.05). For kidney yang deficiency pattern/syndrome, the total effective rate was 79.2% (19/24) in the plaster therapy group, better than 76.9% (22/26) in the nasal spray group (P<0.05).

CONCLUSIONThe dog days moxibustion plaster therapy achieves definite efficacy on allergic rhinitis at the acupoints selected based on the differentiation of different patterns/syndromes and the efficacy is better than beclometasone dipropionate aqueous nasal spray.

Acupuncture Points ; Administration, Cutaneous ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Child ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Rhinitis, Allergic ; drug therapy ; therapy ; Yang Deficiency ; drug therapy ; Young Adult

Objective To explore the investigate of X-chromosome-linked inhibitor of apoptosis pro- tein(XIAP)and its effect on chemotherapeutic sensitivity in bladder carcinoma.Methods Using immu- nohistochemistry methods,the expression of XIAP was evaluated in 47 bladder carcinomas and 6 normal bladder tissues.The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418.Cellular XIAP mRNA level was detected by RT-PCR.The apoptosis of T24 cells was induced by low-dose of mitocycin C(0.005 mg/ml and 0.05 mg/ml,respectively).The in vitro cellular growth activities were assayed by MTT color imetry;and the apoptosis rate was assayed by TUNEL methods. Results The expression rate of XIAP was 78.7%(37/47)in bladder carcinoma samples,with no corre- lation with carcinoma stages and grades(P＞0.05).XIAP mRNA level in transfected T24 ceils was signifi- cantly increased by 3.8 times.Treated with 0.005 mg/ml and 0.05 mg/ml of mitomycin C,the growth rates of XIAP transfected T24 cells were increased [(11.60?0.25)% and(16.51?0.87)% ,respectively,P＜0.05];and the apoptosis rates were decreased [(10.1?0.2)% and( 11.9?0.2)% ,respectively,P＜0.05]compared with those in control cells.Conclusions XIAP is highly expressed in humun bladder car- cinoma samples.Overexpression of XIAP in T24 cells results in decrease in bladder carcinoma cell apoptosis induced by MMC,which may decrease the chemotherapeutic sensitivity of T24 cells.

Objective To find out the iodine nutritional status and thyroid function of pregnant and lactating women in the urban areas of Wuwei City of Gansu Province,and to provide a scientific basis for iodine supplementation.Methods Blood and urine samples of pregnant (52 persons) and lactating women (59 persons) were collected in 2009.Urinary iodine content was measured with arsenic cerium catalytic spectrophotometric.Serum free three triiodothyronine(FT3),free thyroid hormone(FT4),triiodo thyronine(TT3),serum total thyroxine (TT4) and thyroid-stimulating hormone (TSH) were detected with direct chemiluminescence immunoassay.Results The medians urinary iodine of pregnant and lactating women were 274.68,217.88 μg/L.The rates of low urinary iodine (pregnant women below 150 μg/L,lactating women lower than 100 μg/L) were 9.62％ (5/52) and 6.78％ (4/59).Serum TT3,TT4 levels in pregnant women[(2.48 ± 0.59),(132.18 ± 33.36)nmoL/L] were higher than that in the lactating women[(2.16 ± 0.49),(108.79 ± 28.36)nmol/L,t =-3.123,-3.971,all P ＜ 0.05].Thyroid dysfunction incidence rates of pregnant and lactating women were 17.31％ (9/52) and 8.47％ (5/59).Thyroid dysfunction was mostly subclinical hypothyroidism.Conclusions Overall iodine nutrition of pregnant and lactating women is in good condition,some individuals have the trend of hypothyroidism.It is necessary to carry out routine monitoring of urinary iodine and thyroid function.

To automatically infer the patterns of vessel structure such as the distal ends, segments, bifurvessel structures, and crossing of two vessels in X-ray angiographic images, a novel method is presented based on Gabor filter and circle detector. The method can cope with varying vessel curvature and intensity feature occur along the longitudinal vessel direction. The present study can facilitate 2-D quantitative description of vessel tree and 3-D vessel reconstruction, and provide an elementary clue for the diagnostics. The proposed method has been successively applied to both synthetic images for validation purposes and the actual angiographic images, which yielded encouraging results.
Algorithms ; Angiography ; methods ; Artifacts ; Blood Vessels ; anatomy & histology ; pathology ; Humans ; Image Enhancement ; methods ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; Pattern Recognition, Automated ; methods

OBJECTIVETo investigate the effect of PDTC (inhibitor of NF-κb) on apoptosis of human gastric cancer cell line SGC-7901 induced by tumor necrosis factor α (TNF-α) and explore the related mechanisms.

METHODSAfter the treatment with different concentrations of PDTC, TNF-α or PDTC combined with TNF-α on gastric cancer cell line SGC-7901, the growth inhibition of SGC-7901 was measured by MTT assay. Hoechst was used to assess SGC-7901 cell apoptosis. The protein expressions of survivin and caspase-3 were detected by Western blot assay.

RESULTSThe growth inhibition rate of SGC-7901 induced by PDTC (15, 30, 60, 100 μmol/L) was (12.14±0.91)%, (20.00±1.11)%, (37.63±1.01)% and (41.46±1.07)%. Different concentrations of PDTC all inhibited the growth of SGC-7901 significantly (all P<0.01), The growth inhibition rate of SGC-7901 induced by 25 mg/L TNF-α was (2.38±0.67)%, which could not significantly inhibit the growth of SGC-7901 [control (1.50±0.81)%], while TNF-α of 50, 100, 150 mg/L could inhibit the growth of SGC-7901 significantly [(4.53±0.85)%, (4.43±0.70)% and (4.74±1.07)%, all P<0.05]. PDTC (15 μmol/L) combined with TNF-α (25, 50, 100, 150 mg/L) significantly increased the cell growth inhibition rate compared with TNF-α alone or PDTC 15 μmol/L alone (all P<0.01). Hoechst assay showed that 100 mg/L TNF-α, 15 μmol/L PDTC and combination of above two all induced cell apoptosis (P<0.01), and the combination group had significantly higher percentage of cell apoptosis (P<0.01). Survivin protein was significantly down-regulated in combination group as compared with single TNF-α (100 mg/L) group, but was not significant down-regulated as compared with single PDTC (15 μmol/L) group. Caspase-3 protein expression was significantly increased in combination group as compared with other two groups.

CONCLUSIONPDTC can enhance the cell apoptosis induced by TNF-α, which may be associated with the blocking of TNF-α-activated NF-κB signaling pathway by PDTC, the down-regulation of survivin expression, and up-regulation of caspase-3 expression.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Neoplastic ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; NF-kappa B ; antagonists & inhibitors ; Proline ; analogs & derivatives ; pharmacology ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology ; Thiocarbamates ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology