Objective To investigate the relationship between V-raf murine sarcoma viral oncogene homolog B1 (BRAF)v600E mutation and radioactive iodine (RAI) uptake in distant metastases from papillary thyroid cancer(PTC).Methods From January 2011 to December 2012,40 PTC patients (21 males,19 females,average age 39.8 years) with distant metastases were recruited and divided into mutation group and wild group according to the BRAFv600E mutation in primary lesions.The clinical,pathological and serological differences were compared between the two groups.The relationship between BRAFv600E mutation and RAI uptake capability in distant metastases from PTC,as well as its relationship with Tg change after 131I treatment were investigated.Statistical analysis was performed with two-sample t test,x2 test or Fisher exact test.Results The BRAFv600E mutation rate was 30.0％ (12/40) in patients with metastases from PTC.There was no significant difference in clinical,pathological and serological features between mutation group (n =12) and wild group (n=28; t:from-0.533 to 1.728,x2:from-1.951 to 1.088,all P＞0.05).Twelve PTC patients had no RAI uptake in the distant metastases,of which 10 belonged to mutation group (83.3％,10/12) and 2 belonged to wild group (7.1％,2/28; x2=19.734,P＜0.05).BRAFv600E mutation group was more likely to have no RAI uptake in the distant metastases.Tg change after 131I treatment in 30 patients were analyzed.In the wild group,Tg level decreased in 66.7％ (14/21) patients,stabilized in 19.0％ (4/21)and increased in 14.3％ (3/21)patients.While there was no decrease of Tg in the mutation group (0/9).Two patients had increased Tg level and 7 patients (with no RAI uptake) kept stable in mutation group.Conclusions Due to poor RAI uptake capability in PTC patients with BRAFv600E mutation,both primary and metastatic sites may have poor response to 131I treatment.Molecular detection of BRAFv600E mutation might be helpful for choosing PTC with distant metastases and predicting the effect of 131 I treatment.

Objective To investigate the change of serum Tg levels of DTC patients with positive stimulated Tg (Tg ≥ 10.00 μg/L),negative 131I-diagnostic whole body scan(Dx-WBS) and no distant metastasis 6 months after initial 131I therapy.Methods Fifty-six DTC patients (20 males,36 females,average age 43.11 (21-70) y) with intermediate or low risk of recurrence according to American Thyroid Association (ATA) guideline were enrolled into the retrospective study.All patients were grouped according to stimulated Tg level after initial 131I therapy:group with positive Tg (Tg+ group,n =19) and group with negative Tg (Tgˉ group,n=37).Changes of suppressed Tg at 1 year and 2.5 years (Tg1ysup and Tg2.5ysup) after initial therapy were compared between the two groups.Serum TSH level,TgAb level,neck ultrasound and chest CT results were also evaluated.The two-sample t test and x2 test were used for statistical analysis with SPSS 17.0.Results Stimulated Tg and Tglysup levels in Tg+ group were remarkably higher than those in Tgˉ group:(24.27±4.10) μg/L vs (2.73±3.01) μg/L,t=7.191,P＜0.05(6 months after initial 131I therapy) ; (2.21±0.55) vs (0.48±0.10) μg/L,t=3.102,P＜0.05(1 year after initial 131I therapy),respectively.In Tg+ group,suppressed Tg level decreased with time in 68.4％ (13/19) of patients,of whom the Tg2.5ysup level was much lower than Tglysup level ((0.53±0.15) μg/L vs (1.38±0.50) μg/L).Tg2.5sup level in Tg+ group became comparable to that in Tgˉ group ((1.44±0.52) μg/L vs (0.38±0.07) μg/L; t =2.001,P＞0.05).In each group,one case of recurrence with suppressed Tg of 1.4 μg/L and 0.1 μg/L respectively,was observed using neck ultrasound after 2 years of follow-up.Conclusions Serum Tg levels decreased with time for Tg+/131I-Dx-WBS-DTC patients with intermediate or low risk of recurrence.It might not be necessary to follow up these patients with Tg and 131 I-DxWBS after 6 months of initial 131I therapy.

OBJECTIVETo observe the neurological protective effects of progesterone (PROG) on focal cerebral ischemia/reperfusion injury in rats and to explore its possible mechanism.

METHODSOne handred and twenty male SD rats were divided into three groups randomly: sham-operated group, middle cerebral artery occlusion ( MCAO ) group and PROG + MCAO group( n = 40). The right temporary MCAO model was established by the line-embolism method. The PROG + MCAO group rats were according to 8 mg/kg intraperitoneal injection PROG, after that 30 min, the rats were suffered ischemia/reperfusion. After rats were suffered ischemia for 2 h and reperfusion 0, 24, 48, 72 h stress, the nervous functional defect degree were evaluated by longe scoring, and the expression of two-pore domain K channel 3 (TASK3) mRNA in brain tissue were detected by the real-time PCR.

RESULTSPROG (8 mg/kg) could significantly reduced the nervous functional defect degree in rats after ischemia/reperfusion 24, 48, 72 h (P < 0.05). The results of real-time PCR showed that the TASK3 mRNA expression in the brain tissue at all time points significantly decreased in MCAO group compared with sham-operated group (P < 0.05). However, compared with MCAO group, the expression of TASK3 mRNA in brain tissue at all time points dramatically increased in PROG + MCAO group (P < 0.05).

CONCLUSIONPROG can improve the nervous functional defect degree after focal cerebral ischemia/reperfusion injury in rats, and the mechanism might be associated with up-regulating the expression of TASK3 mRNA in brain tissue.

Animals ; Brain ; drug effects ; pathology ; Brain Ischemia ; drug therapy ; Infarction, Middle Cerebral Artery ; Male ; Neuroprotective Agents ; pharmacology ; Potassium Channels, Tandem Pore Domain ; metabolism ; Progesterone ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Reperfusion Injury ; drug therapy

AIM: Gene transduction with a recombined murine stem cell retroviral vector has been investigated to find an effective way of gene transduction and to offer theory and experimental basis for the recombined murine stem cell retroviral vector used for gene transduction. METHODS: 1. Construction of retrovirus vector: EC1-4 gene (repeats 1-4 of cadherin-5 extracellular domain) and mutant (Ser 222A) MEK1 gene were cloned into retrovirus vector pMSCV after cut by Bgl Ⅱ and EcoR 1 restriction endonuclease. 2. Obtaining CD41 + cells and cell culture: Cells expressing CD34 + from cord blood were isolated. The inducement of cells expressing CD41 from CD34 + cells was performed by using TPO and cells CD41 + were selected by FACS. NIH 3T3 cells were cultured in high sugar DMEM medium and U937 in RPMI 1640 medium. UT7 cells which is cytokine-dependent cell line were grown in Iscove's modified Dulbeco's medium supplemented by GM-CSF. 3. Determination of viral titers: Retroviral vectors were transferred to packaging cell line 293. Retroviral containing supenatant was collected after transfection. The viral titers was tested on infection of NIH 3T3 cells by FACS analysis. 4. Western blot: Transduced CD41 +,UT7,U937 and MDA-MB-435 cells were analyzed by western blot to examine expression of transduced genes. RESULTS: A packaging cell line 293 produces high-titer MEK1 pMSCV retroviruses (3.1?10 7) and EC1-4 pMSCV retroviruses (1.0?10 8). With 8-folds dilution retroviruses,60.73% GFP positive cells have been obtained in MEK1 pMSCV transduced UT7 cells,72.56% in U937 cells and 30.57% in CD41 + cells,respectively. GFP positive cells have reached up 97.54 % in EC1-4 pMSCV transducted MDA-MB-435 cells. Phosphorylated MEK1 has been decreased in experiment group when TPO has stimulated CD41 + and UT7 cells or serum has stimulated U973 cells. This indicates that is a dominant negative effect of mutation MEK gene. EC1-4 gene transduced MDA-MB-435 cells have expressed EC1-4. CONCLUSION: The recombined murine stem cell retrovirus can effectively mediate gene transduction of CD41 +,UT7,U937 and MDA-MB-435 cells,and transduced genes can be stably expressed.

The optimization on liquid fermentation and purification of the fibrinolytic enzyme from Stenotrophomonas maltophilia(DR-929) was investigated.The results showed the best fermentation are amidulin 2.0%,soya flour 1.0%,yeast extract 0.5%,NaCl 1.0%,CaCl2 0.02%,MgSO4 0.05%,inoculum of 36 hours,fermental time 4d,initial pH 8.0 or 9.0,temperature 25℃,volume of media 30ml,volume of inoculum 5% or 6%.The purification process includes the following steps: removing cells by the centrifugation,25%~70% saturation ammonium sulfate precipitation,HIC with Phenyl FF(high sub),IEC with Q-Sepharose FF,gel filtration chromatography with Superdex 75.SDS-PAGE electrophoresis was used to examine the purification effect,and the results indicated that homogeneous strap in SDS-PAGE and has a molecular weight around 28.3kDa.The purification factor and activity recovery of the fibrinolytic enzyme are 271.5 and 24.5%,respectively.

A rapid LAMP detection method with primers designed on genus-specific region identified in the gyrA gene was established in this assay. All four Campylobacter jejuni from different sources were detected positive and fourteen non Campylobacter bacteria were negative, which shows excellent specificity of the primers. Compared with plate count and PCR method, the LAMP method and the PCR method had equal sensitivity, which were three orders of magnitude higher than plate count. In this assay, we also found out that the treatment of DNase could reduce the dead bacteria DNA effectively. The LAMP detection on chicken indicated relatively good result on detection of Campylobacter jejuni combining with treatment of DNase.

Objective To investigate associations of UC with the polymorphisms of TRAIL receptors.Methods From January 2008 to December 2012, 380 consecutive UC patients [215 males and 165 females, the average age was (42.63 ±14.61) years] as well as 539 sex-and age-matched healthy individuals [290 males and 249 females, the average age was (41.29 ±15.86) years] were recruited from four large scale comprehensive hospitals in Wenzhou city.Five single nucleotide polymorphisms of DR4 (rs20575, rs13278062), DR5(rs1047266), DcR2(rs1133782) and OPG (rs3102735) were detected by a SNaPshot technique.Distributions of mutant alleles and genotypes for targeted polymorphisms in TRAIL receptors were analyzed by Chi-square test or Fisher′s exact test. By means of unconditional Logistic regression analysis, it evaluated associations between the polymorphisms and the risk of UC attack as well as the clinical features of UC patients.Furthermore, an unconditional Logistic multiple regression analysis was employed to investigate the independent risk factors of UC and their multiplicative interaction effects on UC.Results The frequencies of mutant allele (G) and genotype (CG+GG) of DR4(rs20575) were higher in UC patients than those in the controls (3.55%vs 1.95%,χ2 =4.512, P=0.034;6.58%vs 3.71%,χ2=3.938, P=0.047, respectively).However, the frequeucies of mutant allele ( A) and genotype ( GA+AA) of DcR2(rs1133782) were decreased in UC patients compared to the controls(6.18%vs 9.09%,χ2=5.183, P=0.023; 11.32% vs 17.44%, χ2 =6.589, P=0.010, respectively).The frequencies of mutant allele (T) and homozygote (TT) of OPG(rs3102735) were significantly higher in UC patients than in the controls (86.32% vs 81.54%, χ2 =7.385, P=0.007;75.26% vs 66.98%, χ2 =7.346, P=0.007, respectively) .Furthermore, the genotype (GG) of DcR2 (rs1133782) was found to be the independent risk factor for UC attack (OR=4.937, 95%CI:2.320-10.504, P<0.001).Moreover, the (GG) of DcR2(rs1133782) and (CC) of DR4(rs20575) had an interactive effect on UC (OR=0.322, 95%CI:0.164-0.633, P=0.001).The same conclusion was drawn for the ( GG) of DR4( rs20575) and (TT) of OPG(rs3102735) (OR=1.580, 95%CI:1.165-2.144, P=0.003).Conclusions The genetic polymorphisms of DR4 ( rs20575 ) , DcR2 ( rs1133782 ) and OPG ( rs3102735 ) were associated with UC. The mutation of DcR2(rs1133782) might play a protective role in UC.Moreover, the DcR2(rs1133782) and DR4(rs20575) gene had a collaborative effect on UC.So did the DR4(rs20575) and OPG(rs3102735) genes.

BACKGROUND: Silibinin has broad pharmaceutical effects, such as anti-free radicals, anti-lipid peroxidation, anti-lipoid oxidase, anti-glutathione (GSH) depletion, anti-neoplastic and serum lipid-lowering effects. Clinically, silibinin is often used in treating alcoholic liver disease. OBJECTIVE: To investigate the pharmacological mechanism of silibinin for alcoholic fatty liver in rats. DESIGN: Randomized and controlled study.SETTING: Guangzhou Hospital of Traditional Chinese Medicine.MATERIALS: The experiment was conducted at the Animal Experimental Laboratory of Guangdong Pharmaceutical Institute from August to October 2003. Totally 57 SD rats, without unusual bacteria, weighting (150±10)g and of either gender, were selected. Yiganling tablets containing 38.5 mg silibinin were produced by Zhuzhou No.3 Pharmaceutical Factory (Batch No. 20020808).METHODS: Among the 57 SD rats, 18 rats were regarded as normal control group. Rats in normal control group were administered with normal saline by gavage, and fed with normal food and distilled water in place of alcohol for 10 weeks. Rats in model group and silibinin group were fed with high-calorie food and 100 mL/L alcohol for 6 weeks to establish model of rat alcoholic fatty liver. The other rats were divided into model control group (n=18) and silibinin group (n=21). Rats in model control group were treated with distilled water while those in silibinin group were treated with 100 mg/kg silibinin. Meanwhile, 100 mL/L ethanol and hyperalimentation feed were given for 4 weeks. After animals were killed, TG, SOD, GSH and MDA levels were measured with liver suspension.MAIN OUTCOME MEASURES: Contents of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), total cholesterol (TC), triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), tumor necrosis factor (TNF)-α , and transforming growth factor (TGF)-β1.RESULTS: All the 57 rats entered the final analysis. Silibinin could inhibit the activities of serum AST, ALT and AKP [(2 550.5±400.1), (533.4±100.0), (2 217.1±750.2)nkat/L], and the differences were significant as compared with those in model control group [(3 600.7±666.8), (800.2±100.0), (2 900.6±1 333.6) nkat/L, P ＜ 0.05-0.01]. Contents of TG, LDL-C, TNF-α and TGF-β1 in silibinin group [(1.8±0.8), (0.17±0.04), (6.66±1.38), (24.1±4.1) mmol/L] were lower than those in model group [(2.8±1.4), (0.20±0.05), (7.81±1.06), (28.8±6.3) mmol/L] with significant differences (P ＜ 0.05-0.01). Silibinin could increase the content of HDL-C but decrease the contents of TG and MDA (P ＜ 0.05-0.01), and improve SOD activity as well as hepatocyte and fatty degeneration (P ＜ 0.01).However, it had no obvious effect on the content of reduced estathion (P ＞ 0.05).CONCLUSION: Silibinin can inhibitthe formation of alcoholic fatty liver in rats. The pharmacological mechanism of silibinin may involve anti-oxidation, removing free radicals, inhibiting lipid peroxidation, regulating blood lipid component, reducing fatty sediment in liver, and anti-immunoinflammation and anti-hyperplasia effects.

Objective To screen the arsenic content situation of drinking water in lower reaches of Yellow River and survey the amount of threatened people drinking high arsenic water and the condition of endemic arsenism.Methods Four counties of Yuncheng,Jiaxiang,Dongchangfu and Boxing were selected to colleft the water samples by CroOSS-sectional survey method.The water arsenic content wag determined by semi-quantitative rapid kit.All water samples having arsenic were re-determined by atomic fluorescence spectrometry.And the nurober of threatened people who drinking high arsenic water were investigated.Results In 4765 water wells screened,303 water samples had contained arsenic,arsenic content of 35 samples Was≥0.030 mg/L,12 samples were exceeding the international standard (arsenic content≥0.050 ms/L),they distributed in 3 counties of Dongchangfu,Yuncheng and Jiaxiang.The residents drinking water wells of arsenic content≥0.030 mg/L were surveyed by epidemiological investigation.And in the 28 villages 13 032 residents and 11 Bu8picious patient8 wlere checked out.Conclusion The wells with excesive water arsenic content are existing in the lower reaches of Yellow River and people suspicious of endemic arsenism need to be further identified.

OBJECTIVETo evaluate the clinical effects of laparoscopic occlusion of the internal spermatic vein with Hem-o-Lock clips and retroperitoneal ligation of the spermatic vessel (Palomo procedure) in the treatment of primary varicocele.

METHODSWe included in this study 42 varicocele patients treated by laparoscopic occlusion of the internal spermatic vein with Hem-o-Lock clips (Hem-o-Lock group) and another 38 treated by Palomo procedure (Palomo group). We recorded the operation time, intraoperative blood loss, hospital stay, bowel function recovery time, post-operative complications, as well as such seminal parameters as sperm concentration, sperm motility, sperm abnormality and the percentage of grade a + b sperm 6 months after surgery. We measured the interior diameter and reflux time of the spermatic vein and the ratio of flow velocity to the diameter using color Doppler flow imaging (CDFI), and observed the disappearance and recurrence of clinical symptoms.

RESULTSIntraoperative blood loss was markedly less and the operation time, postoperative hospital stay and intestinal function recovery time significantly shorter in the Hem-o-Lock than in the Palomo group (P<0.05), but there were no statistically significant differences in the incidence of postoperative complications (P>0.05). Six months after surgery, seminal parameters and the results of CDFI were significantly improved in both the Hem-o-Lock and Palomo groups (P<0.05), but with no significant difference between the two (P>0.05). No recurrence was found in either group at 18 months.

CONCLUSIONBoth laparoscopic spermatic vein occlusion with Hem-o-Lock clips and Palomo procedure can improve the semen quality and relieve the clinical symptoms of primary varicocele patients, but the former is even more preferable for its shorter operation time and hospital stay, less intraoperative bleeding, fewer complications, better clinical outcomes, and lower rate of recurrence.

Adult ; Humans ; Laparoscopy ; Male ; Treatment Outcome ; Varicocele ; surgery ; Vascular Surgical Procedures ; methods ; Veins ; surgery ; Young Adult