Lentivirus vectors construction of SiRNA targeting interference GPC3 gene and its biological effects on liver cancer cell lines Huh-7

Objective:To build GPC3 gene short hairpin interference RNA(shRNA)slow virus veclor.observe expression of Huh-7 GPC3 gene in human liver cell line proliferation apoptosis and the effect of GPC3 gene influencing on liver cancer cell growth,and provide theoretical basis for genc therapy of liver cancer.Methods:Hepatocellular carcinoma cell line Huh-7 wsa transfected by a RNA interference technique.GPC3 gene expression in a variety of liver cancer cell lines was detected by fluorescence quantitative PCR.Targeted GPC3 gene seqnences of small interfering RNA(siRNA)PGC-shRNA-GPC3 were restructured.Stable expression cell linse of siRNA were screened and established with the heplp of liposomes(lipofectamine^(TM2000))as carrier transfcetion of human liver cell lines.In order to validate siRNA interference efficiency.GPC3 siRNA mRNA expression was detected after transfection by using RT-PCR and Western blot.The absorbance value of the cells of blank group,untransfection group and transfection group,the cell cycle and cell apoptosis were calculated,and effects of GPC3 gene nn Huh-7 cell proliferation and apoptosis were observed.Results:In the liver cancer cell lines Huh-7 GPC3 gene showed high expression.PGC-shRNA-GPC3 recombinant plasmid was constructde successfully via sequencing validation.Stable recombinant plasmid transfected into liver cancer cell linse Huh-7can obviously inhibit GPC3 mRNA expression level.Conclusions:The targeted GPC3 siRNA can effectively inhibit the expression of GPC3.

Effects of Lanthanum Carbonate on Vascular Calcification in Elderly Maintenance Hemodialysis Patients

Summary： The effect of lanthanum carbonate on abdominal aortic calcification （AAC） in the elderly maintenance hemodialysis （MHD） patients was investigated. Fifty-four cases subjected to routine MHD complicated with skin pruritus admitted to our hospital were selected and randomly divided into case group （n=28） and control group （n=26）. The control group was given routine MHD alone. The case group was given lanthanum carbonate additionally on the basis of routine MHD. The changes of itching degrees at first and third month, and serum calcium, phosphorus, calcium-phosphorus products, intact parathyroid hormone （iPTH） levels and AAC scores at third month after treatments were compared be- tween the two groups. The correlation between calcium-phosphorus products and AAC scores was also analyzed. There was no significant difference in the baseline of blood urea nitrogen （BUN）, serum creatinine （Scr）, uric acid, albumin, hemoglobin, C reactive protein （CRP）, low density lipoprotein （LDL）, high density lipoprotein （HDL）, triglyceride, total cholesterol between case group and control group （P〉0.05 for all）. There was also no significant difference in the baseline itching scores between the case group and the control group （P〉0.05）. At 1st and 3rd month after treatment, the itching scores in the case group were 14.2±3.2 and 10.5±2.3, respectively, which were significantly lower than the baseline and those in the control group （P〈0.05 for all）. At 1 st and 3rd month after treatment, the itching scores in the control group were 23.6v5.9 and 24.8±6.3, respectively, which were significantly higher than the baseline （P〈0.05）. There was no significant difference in the baseline of serum calcium, phos- phorus, calcium-phosphorus products, iPTH levels between the case group and control group （P〉0.05）. At 3rd month after treatment, serum phosphorus, calcium-phosphorus products and iPTH levels in the case group were decreased significantly as compared with the baseline （P〈0.05）, and the serum calcium, phosphorus, calcium-phosphorus products, and iPTH levels were statistically decreased as compared with those in the control group （P〈0.05）. The AAC scores showed statistically significant difference between the case group and the control group （P〈0.05）. The serum phosphorus and AAC scores showed a positive correlation in both two groups. It was suggested that the administration of lanthanum carbon- ate in the elderly MHD patients can effectively relieve itching, and simultaneously reduce serum phos- phorus and iPTH levels, resulting in the attenuation of vascular calcification.

Study on medication laws of traditional Chinese medicine of 340 cases of COVID-19 based on data mining

Objective:To analyze the prescription rules of traditional Chinese medicine for patients with COVID-19 in Wuhan Third Hospital, and summarize the disease characteristics and clinical treatment ideas, in order to learn from the treatment of this disease by traditional Chinese medicine.Methods: This study established a cloud database based on the ancient and modern medical cases to mine the rules of medication with analyses of frequency of drug used, association analysis, cluster analysis, complex network and other methods.Results:Of 340 COVID-19 patients discharged from our hospital, 143 males and 197 females with male: female = 0.73: 1, and there were 167 patients over 60 years old, and the majority of clinical illnesses were mainly middle-aged and elderly people. Most of them had dry cough, anorexia, fatigue, etc. The traditional Chinese medicine for the treatment of COVID-19 involves a total of 204 flavors. The most frequent use is Fuling of 225 times. The treatment of traditional Chinese medicines is mainly involved with flavors such as warm, spicy and sweet. Mainly detoxification, dampness and phlegm, Qingfei Paidu Decoction is a common clinical prescription, of which the core drugs are Huoxiang, Fabanxia, Chenpi, Baizhu, Cangzhu, Fuling, Chaihu, Huangqin, Xingren, and Gancao.Conclusion: Through this study, the patients with COVID-19 in our hospital are mainly middle-aged and elderly people, and the etiology is characterized by "cold, wet, poisonous, and epidemic". Wetness is the most important core of the disease.

Effect of Bijia Jian Pill on inflammatory factors and liver fibrosis indexes in patients with chronic hepatitis B complicated with cirrhosis

Objective: To detect the effect of BiJia Jian Pill on inflammatory factors and liver fibrosis indexes in patients with chronic hepatitis B complicated with cirrhosis. Methods: A total of 94 patients with chronic hepatitis B combined with cirrhosis admitted in the hospital from January 2017 to December 2017 were selected as the subjects, and they were randomly divided into 47 cases in the observation group and 47 cases in the control group. The control group was given Adefovir Dipivoxil Tablets, and the observation group was treated with Bijia Jian pills on the basis of control group. The changes of liver function, inflammatory factors and liver fibrosis indexes were compared between the two groups before and after treatment. Results Before treatment, there was no significant difference in AST, ALT, TBIL and ALB between the two groups. After treatment, the AST, ALT and TBIL of the two groups were all decreased, and the AST, ALT and TBIL in the observation group were lower than those of the control group;the ALB of the two groups increased, and the ALB of the observation group was higher than that of the control group. Before treatment, there was no significant difference in TNF-α, IL-1 βand IL-10 between the two groups. After treatment, the TNF-α and IL-1β of the two groups were decreased, and the TNF-α and IL-1β in the observation group were lower than those of the control group;the IL-10 of the two groups increased, and the IL-10 in the observation group was higher than that in the control group. Before treatment, there was no significant difference in HA, LN, IV-C and PCIII between the two groups. After treatment, the HA, LN, IV-C and PCIII of the two groups were all decreased, and the HA, LN, IV-C and PCIII of the observation group were lower than those of the control group. Conclusion Bijia Jian pills can effectively improve the inflammatory factors and liver fibrosis in patients with chronic hepatitis B complicated with cirrhosis.