Objective To investigate the inhibitory effect of recombinant LIGHT-Fe gene on the proliferation of human esophageal carcinoma cell line Eea109. Methods LIGHT-Fc expression vector was transfected into human esophageal squamous carcinoma cell line Eca109 by using DOTAP liposomal transfection reagents. The effects of LIGHT-Fc gene on the proliferation of esophageal carcinoma cell line Eca109 in vitro were detected by cell growth curve and MTr assay. Forty-five nude mice were equally divided into Eea109/Wt group, Eca109/neo group and Eca109/LIGHT group. Carcinogenesis and pathological expression of the esophageal carcinoma tissues were observed. Results The expressions of LIGHT receptors were detected in Eca109 cells. The proliferation of Eca109 cells was inhibited after trasfecting LIGHT-Fc gene into Eca109 cells. The numbers of tumors generated in Eea109/Wt group, Eca109/neo group and Eca109/LIGHT group were 12, 11 and 5, with statistical significance between Eca109/LIGHT group and the other two groups (X2 =6.652, 4.821, P <0.05). The result of histopatholagical examination indicated that the tissue necrosis appeared significantly in tumors derived from Eea109/LIGHT cells. Conclusions The growth of esophageal squamous carcinoma cell line Eca109 can be suppressed by LIGHT-Fc gene whether in vitro or in vivo.

Objective To investigate the therapeutic effects and the possible mechanism of fecal transplantation on rats with Clostridium difficile-associated pseudomembranous colitis. Methods A total of 40 Sprague-Dawley rats were divided into four groups including the healthy control group, model group, fecal transplant treatment group and vancomycin treatment group. Rats in three experimental groups were subcuta-neously injected with clindamycin phosphate (10 mg), followed by treatment with toxin producing Clostridi-um difficile (ACTT43255) enema 24 hours later. The rats in fecal transplant treatment group and vancomy-cin treatment group were respectively treated with fecal suspension and vancomycin one day after modeling. The rats were fasted for one day after the last administration and then executed. The levels of potassium ion ( K) , sodium ion ( Na) , albumin ( ALB) , white blood cells ( WBC) , C-reaction protein ( CRP) , interleu-kin-1β ( IL-1β) , interleukin-10 ( IL-10 ) , interleukin-12 ( IL-12 ) and interleukin-17 ( IL-17 ) as well as the percentage of neutrophils ( N%) in serum samples were detected. The colon tissue samples were collect-ed for pathology examination. Results The rat model of pseudomembranous colitis was successfully estab-lished by subcutaneous injection of clindamycin in combination with toxin-producing Clostridium difficile (ACTT43255) enema. The signs of intestinal inflammation including serious weight loss, remarkably short-ened colon length and significantly increased colon wet weight index were observed in rats from the model group (P<0. 05). Compared with the rats from model group, the rats received fecal transplant showed sig-nificantly increased levels of K, ALB, IL-10 and IL-10/IL-12 in serum and decreased levels of WBC, N%, CRP, IL-1β and IL-17 (P<0. 05). Conclusion Fecal transplantation was proved to be an effective ap-proach for the treatment of pseudomembranous colitis. The therapeutic mechanism might due to its impacts on serum inflammatory factors.

Objective To establish a new approach based on mathematical morphology that can effectively reduce the drifts in functional magnetic resonance imaging (fMRI) signals. Methods Based on investigation of the characteristic of drifted fMRI signals, a mathematical morphology method for baseline drift correction was presented. Results With both simulated data and real fMRI data, the results of the experiment showed that the mathematical morphology method can effectively correct the baseline drifts. Conclusion Both linear and nonlinear drifts can be removed with the proposed method without any statistical model assumption.

This study was aimed to improve the transfer rate of sinomenine in the intermediate product of Caulis Sinomenii in order to optimize the purification process of intermediate product ofCaulis Sinomenii fromTong-An (TA) injection. The transfer rate of sinomenine and the stability of fingerprints in the intermediate product of Caulis Sinomenii were used as indexes for the investigation on the impact from different pH ofCaulis Sinomenii extract before extraction. The transfer rate of sinomenine was used as an index for the investigation on the impact by different pH of hydrochloric acid created to dry extract solution. The transfer rate of sinomenine was used as an index for the investigation of four separation ways, which included the vacuum filtration, plate and frame filters, high-speed tube separator, and flat direct centrifuge, on the liquid separation of sinomenium acutum acid. The results showed that the pH ofCaulis Sinomenii extract before extraction was 10-11; the transfer rate of sinomenine was the highest in the extraction process and the fingerprints of TA injection was stable. The pH of hydrochloric acid was 2.0-2.5; and the highest transfer rate of sinomenine in acid dissolution process was 92.94%. The high-speed tube separator had the best separation to sinomenium acutum acid-dissolving liquid. The highest transfer rate of sinomenine was 93.34%. It was concluded that the optimized process can effectively improve the transfer rate of sinomenine in the intermediate product ofCaulis Sinomenii. Meanwhile, fingerprints of the product were stable. The process was simple with good repeatability.

Objective To investigate the effect of α-galactosidase A (GLA) gene mutation on cell autophagy and to elucidate its mechanism preliminarily.Methods Two families were diagnosed by ultrastructural pathological examination,GLA gene activity test and GLA gene mutation screening.Mutant type recombinant expression plasmid of two pedigrees (pcDNA3.1-GFP-ex1 (EX1 group),pcDNA3.1-GFP-ex3 (EX3 group)) and wild type recombinant expression plasmid of GLA (pcDNA3.1-GFP-GLA,GLA group) were constructed.Hela cell line (control group) was transiently transfected with recombinant expression plasmid according to lipofectin transfection.The relative gene expression of Beclin-1 was measured with real-time PCR,and protein expression level of LC3-Ⅱ/LC3-Ⅰ,Beclin-1 and P62/SQSTM1 was examined by Western blotting.Results The LC3 protein values of groups EX1,EX3,GLA and control were 1.495 ± 0.064,1.490 ± 0.020,1.285 ± 0.021,1.260 ± 0.042,respectively;P62/ SQSTM1 values were 0.555 ± 0.086,0.480 ± 0.084,0.785 ± 0.439,0.980 ± 0.278,respectively;Beclin-1 mRNA 2-△Ct values were 0.011 ±0.003,0.008 ±0.002,0.005 ±0.001,0.003 ±0.001,respectively;Beclin-1 protein values were 1.178 ±0.098,1.209 ±0.092,0.931 ±0.100,0.796 ±0.184,respectively.Compared with the wide type group,the level of LC3-Ⅱ/LC3-Ⅰ protein was significantly higher in the mutant type groups(t =5.118,4.984;P =0.007,0.008),though no statistically significant difference was found in the expression levels of P62/SQSTM1 (t =1.052,1.400;P =0.323,0.199).Besides,the expression levels of Beclin-1 mRNA (t =3.800,2.445;P =0.005,0.040) and protein (t =2.424,2.729;P =0.042,0.026) were significantly higher in the mutant type groups.Conclusions GLA gene mutation can induce cell autophagic dysfunction,and signaling pathway of autophagic activation may be Beclin-1 dependent.

Objective:To explore the association of the genes GSTM1 and GSTT1 with oligozoospermia infertility.Methods:PCR technique was used to analyze the GSTM1 and GSTT1 genes polymorphisms in 75 with oligospermia infertility and 36 healthy individuals of Zhuang population from Guangxi Baise area,and then the possibility function of GSTM1 and GSTT1 genes in human oligozoospermia were studied.Results:Analyses of the polymorphisms in GSTM1 and GSTT1 genes showed that GSTM1 defect or combined defects of both GSTM1 and GSTT1 was found more frequent in patients with infertile oligospermia than in the healthy control.Conclusion:GSTM1 and GSTT1 genes may have modulating effects on human spermatogenesis,whose mechanism needs further studies.

Objective To evaluate and summarize a new way of forward intensity modulated radia- tion therapy (IMRT) plan for nasopharyngeal carcinoma with 3Dmulti-leaf collimator (MLC) planning sys- tems is practised routinely in our department.Methods From September 2000 to July 2003 and November 2005 to March 2006,78 patients with nasopharyngeal carcinoma who were treated with eonformal radiothera- py used CT simulation for localizing,then doctors supervised the delineation of the planned tumor volume (PTV),gross tumor volume (GTV) and other sensitive organs on ACQsim workstation,and then sent the CT imagines to ELEKTA Precise Plan by DICOM RT Ethernet.Then,the physicists take over the responsibility of all directions of beam projection according to these organs and PTV shape,and completed the IMRT plan by manual correction making and the optimization of fields or segments shaped with forward 3D planning sys- tem.Results To analyse all the patient's dose distributions taking conformal radiotherapy in our hospital, we found the dose distribution and DVH data excellent,even better than the inverse planning of nasopharyn- geal carcinoma.Conclusions Intensity modulated radiation therapy (IMRT) is an advanced radiotherapy technique.A very good IMRT plan of nasopharyngeal carcinoma by forward planning can be obtained with 3D multiple leaf collimator (MLC) planning system.This planning method is more flexible,but the radia- tion physicists should be very much experienced in professional skill.

Four primers against the genes encoding(cpa,cpb,etx,and iA) four major toxins(?,?,?,?) of Cl. perfringens were designed and the colony multiplex PCR of identification and genotyping of Cl. perfringens strains were developed. Cl. perfringens reference strains stored in china institute of veterinary drug control including A,B,C,D and E genotyping were genotyped using the colony muitiplex PCR assay. The expected sequences were obtained successfully by the colony multiplex PCR assay. But the sequences were not obtained from Cl. novyi,Cl. septicum and Cl. tetani. The expected sequences were obtained from Cl. perfringens individual colony diluted to 100 times with 0.85% saline solution.13 Cl. perfringens strains isolated from diferent animals were genotyped using the colony multiplex PCR assay,and the results were comparaed with the results of toxins neutranization test in mice. The two assays showed good accordance. These results showed that the development of the colony multiplex PCR is very important for early and fast identification and genotyping of Cl. perfringens in china.

Objective To evaluate myocardial apoptosis with 99Tcm-C2A-GST myocardial imaging using the recombined C2A domain of Synaptotagmin Ⅰ by gene engineering. Methods ( 1 ) The C2A gene was inserted into the prokaryotic glutathione S-transferate (GST) fusion protein expression plasmid pGEX-6P-1. The recombinant plasmid was transformed into E. coli BL21. C2A-GST fusion protein was purified after BL21 was induced with isopropyl-β-D-1-thiogalactopyranoside (IPTG). (2)The activity of fusion protein was identified by cell binding test with fluorescein-5-isothiocyanate (FITC)-C2A-GST. (3) The C2A-GST fusion protein was labeled with 99Tcm using 2-iminothiophene hydrocoride method. Radiochemical purity was determined with thin layer chromatography. (4)99Tcm-C2A-GST (7.4 MBq) was injected to ischemia-reperfusion rat models through tail vein. The image was acquired with SPECT at 1 h after injection, and then hearts were removed, rinsed with saline and dyed with triphenyl tetrazolium coride (TTC). The ischemic myocardium was separated from the viable myocardium and was weighted. Its radioactivity was measured by gamma counting. The difference of uptake of radiotracer between ischemic myocardium and normal myocardium was compared using percentage activity of injected dose per gram of tissue ( % ID/g) with standard deviation. SPSS 12.0 and t-test were used for data analysis. Results ( 1 ) C2A-GST fusion protein wassuccessfully expressed and its relative molecular weight was 3.8 × 104. (2) FITC-C2A-GST binding to apoptotic cells could be observed by fluorescent microscopy. (3) The radiochemical purity of 99Tcm-C2A-GST was (98.90 ±0.43)%. (4)The imaging studies showed that there was focal uptake of radioactivity in the ischemic myocardium. In vitro uptake of 99Tcm-C2A-GST was (2.41 ±0.32) % ID/g by the ischemic myocardium, however 99Tcm-C2A-GST-N-hydroxysuccinimide (C2A-GST-NHS) was (0. 82 ± 0. 24) % ID/g. There was statistically significant difference between those two groups (t = 10. 6, P ＜0.01 ). Conclusion The C2A domain of Synaptotagmin Ⅰ expressed by gene engineering can be used as the tracer for noninvasive detection of ischemic myocardium in the ischemia-reperfusion rat model.

In order to effectively remove the invalid impurities in Tongan injection, optimize the optimal parameters of the impurity removal technology of liquid mixing process, in this paper, taking Tongan injection as the research object, with the contents of celandine alkali, and sinomenine, solids reduction efficiency, and related substances inspection as the evaluation indexes, the removal of impurities and related substances by the combined process of refrigeration, coction and activated carbon adsorption were investigated, the feasibility of the impurity removal method was definited and the process parameters were optimized. The optimized process parameters were as follows: refrigerated for 36 h, boiled for 15 min, activated carbon dosage of 0.3%, temperature 100 degrees C, adsorption time 10 min. It can effectively remove the tannin, and other impurities, thus ensure the quality and safety of products.
Adsorption ; Charcoal ; chemistry ; Drug Compounding ; instrumentation ; methods ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Quality Control