Establishment of transformation system of Claviceps purpurea Cp-1 strain / 药学学报

Claviceps pururea Cp-1 strain established in our lab is capable of producing variety of bioactive ergot alkaloids, and is broadly used by the pharmaceutical companies. To engineer the strain genetically for the production of specific ergot alkaloid, an effective transformation system must be set up first. However, the reported transformation system is not suitable for this strain due to different genetic backgrounds and the heterogeneity of Claviceps. Thus, in this paper, the hyphae of Cp-1 strain were used to prepare protoplasts by lywallzyme. The formation of protoplasts was investigated under different concentrations and incubation time of enzyme. The strain was tested for sensitivity to several antibiotics at different concentrations. Finally, the genetic transformation system of Cp-1 strain was established. The results suggest that protoplasts were formed efficiently by using 1% lywallzyme at 25℃ for 2 h. Transformants were obtained by PEG mediated protoplast transformation of Cp-1 strain with plasmid pAN7-1, using 1.5 mg·mL-1 hygromycin B as the selective marker. The exogenous gene in the plasmid pAN7-1 was integrated into the genome of Cp-1 strain transformant as demonstrated by PCR result. This study laid an important foundation for genetic manipulation of Cp-1 strain.

Message RNA expression of LUNX, CK19 and CEA genes in NSCLC with micrometastasis in lymph nodes / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the correlation of mRNA expression level of three cancer-associated genes-LUNX mRNA, CK19 mRNA and CEA mRNA with metastasis in lymph nodes and histopathological staging in non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>Fifty-six tumor tissue samples and 103 regional lymph node samples were obtained from 56 patients with NSCLC, and another 35 lymph node samples as control from 15 patients with benign pulmonary diseases. The mRNA expression of LUNX, CK19 and CEA genes was detected in these samples by semi-quantitative RT-PCR analysis (reverse transcriptase polymerase chain reaction), meanwhile, all lymph nodes were also examined by conventional pathological method.</p><p><b>RESULTS</b>mRNA expression of LUNX, CK19 and CEA genes in the regional lymph nodes of NSCLC was significantly higher than that in those of benign lung diseases (P < 0.05). Compared with conventional pathological method, RT-PCT was more sensitive (P < 0.05). No significant correlation was found between positive mRNA expression of LUNX mRNA and CK19 mRNA in the lymph nodes and histopathologic type of lung cancer (P > 0.05). But positive expression rate of CEA mRNA in the lymph nodes from adenocarcinoma patients was significantly higher than that in these from squamous cell carcinoma and other types of NSCLC (P < 0.05). The expression level of LUNX mRNA in the lymph nodes was positively correlated with TNM stages.</p><p><b>CONCLUSION</b>LUNX mRNA and CK19 mRNA may serve as a molecular marker for detection of lymph node micrometastasis in patient with non-small cell lung cancer, but LUNX mRNA is superior to CK19 mRNA in both sensitivity and specificity.</p>

Anti-nasopharyngeal carcinoma effect in vivo and in vitro of Avastin and adenovirus-thymidine kinase suicide gene / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the effect of Avastin and adenovirus-thymidine kinase/ Ganciclovir (Ad-TK/GCV) suicide gene system on nasopharyngeal carcinoma in vivo and in vitro.</p><p><b>METHODS</b>The expression of vascular endothelial growth factor (VEGF) by CNE1 cell line was detected by VEGF ELISA. The effect of Avastin and Ad-TK/GCV on CNE1 cell was detected by methyl thiazolyl tetrazolium (MTT) assay. Flow cytometry analysis and Hoechst 33342 staining were adopted to explore the killing mechanism of Ad-TK/GCV. The nude mice models of CNE1 cell xenografts were established. After intra-tumoral injection of PBS, Avastin, Ad-TK/GCV, Ad-Lac-Z/GCV and Ad-TK/GCV + Avastin, tumor volume was measured and tumor inhibitory rate was calculated. Then the tumors were removed and subjected to histological examination.</p><p><b>RESULTS</b>CNE1 cells could produce VEGF. Avastin had no direct effect on CNE1 cells. The killing effect of Ad-TK/GCV increased with the increase of Ad-TK multiple of infection and the prodrug concentration, which was enhanced by the existence of bystander effect. Compared with control group, the death cell rate (P = 0.000) and apoptosis cell rate (P = 0.000) had significant difference. The study in vivo showed the tumors treated with Avastin, Ad-TK/GCV and Ad-TK/GCV + Avastin grew slowly compared with control. Tumor volume of treated groups was significantly smaller than that of control (all P < 0.05 or P = 0.000). Tumor weight of treated groups was significantly lower than that of control (all P = 0.000). The histological examination showed local necrosis in Ad-TK/GCV group and Ad-TK/GCV + Avastin group, poor angiogenesis in Avastin group and Ad-TK/GCV + Avastin group.</p><p><b>CONCLUSIONS</b>Avastin had no direct effect on CNE1 cells in vitro. Ad-TK/GCV suicide gene system killed NPC cells by inducing cell necrosis and apoptosis, which could be enhanced by the existence of bystander effect. Avastin and Ad-TK/GCV suicide gene system could inhibit the growth of NPC CNE1 cell xenografts. Combination therapy had a synergic effect.</p>

Application of UV similarity in quality control of Chinese medical injection / 中国中药杂志

<p><b>OBJECTIVE</b>To propose an algorithm of the similarity between UV-spectra for controlling the quality of Chinese medical injection.</p><p><b>METHOD</b>The similarity is calculated base on the data of two UV-spectrum curves, thus the sample quality is evaluated according to the average value of the similarity of the UV-spectra between sample with each of standard samples; the proposed algorithm of the similarity is compared with cosine of vectorial angle and relative coefficient; the proposed method for quality evaluation of traditional Chinese injection is discussed with the method according to the chromatographic fingerprint similarity.</p><p><b>RESULT</b>The calculated similarity with the proposed method can be sensitive to show the difference between UV-spectra. This proposed method for quality evaluation was applied for assessing quality of 14 batches of Xiangdan injection and the results well coincident with sample quality.</p><p><b>CONCLUSION</b>The proposed method can be used to evaluate the quality of the Chinese medical injection according to the similarity of two UV-spectrum curves.</p>

Effect of different iodine intake on schoolchildren's thyroid diseases and intelligence in rural areas / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Reports are increasingly appearing on the side effects caused by excessive iodine intake. Our objective was to find out whether iodine excess would impair the thyroid function and intelligence of schoolchildren in rural areas of China.</p><p><b>METHODS</b>A comparative epidemiological study was made on thyroid function and intelligence of the schoolchildren in the areas of low, moderate or excessive intake of iodine. In the area of low intake of iodine (Panshan, Liaoning province, median urinary iodine (MUI) was 99 microg/L), of moderate intake of iodine (Zhangwu, Liaoning Province, MUI was 338 microg/L) and of excessive intake of iodine (Huanghua, Hebei Province, MUI was 631 microg/L). The numbers of schoolchildren from each area selected to take part in a Chinese version of Raven's Test were 190, 236 and 313, respectively, and then 116, 110 and 112 of them were tested for thyroid function, thyroid autoantibody (TAA) and urinary iodine (UI).</p><p><b>RESULTS</b>There were no significant differences in the incidences of overt hyperthyroidism, subclinical hyperthyroidism and overt hypothyroidism in Panshan, Zhangwu and Huanghua. But significant differences were found in the incidences of subclinical hypothyroidism (P = 0.001) in these three areas. The incidences of subclinical hypothyroidism in Huanghua and Zhangwu were 4.76 and 3.37 times higher than that in Panshan. TAA were negative in all the schoolchildren with subclinical hypothyroidism except for one. No significant difference was found among the rates of thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TGAb) in these three areas. Mean serum thyroglobulin (TG) value of Huanghua was markedly higher than those of the other two (P = 0.02). Mean serum TG value of Zhangwu was higher than that of Panshan but the difference was not significant. Mean IQ value of the schoolchildren in Huanghua was markedly higher than that for Zhangwu (P = 0.001). Mean IQ value of the schoolchildren in Panshan was lower than that of Huanghua and higher than that of Zhangwu but, again, the differences were not significant.</p><p><b>CONCLUSIONS</b>The increase of iodine intake may increase the risk for schoolchildren of subclinical hypothyroidism. In the area of iodine excess, most of the subclinical hypothyroidism cases are not of autoimmune origin. No obvious effect of excess iodine was found on mental development of schoolchildren.</p>