1.Herbal Textual Research on Bletillae Rhizoma in Famous Classical Formulas
Dan ZHAO ; Tao ZHOU ; Chaolei LUO ; Dewei GAN ; Lingling LIU ; Chuanzhi KANG ; Zhikun WU ; Xu LI ; Yan FU ; Guoqiong CAO ; Yongping ZHANG ; Chenghong XIAO ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):77-88
In order to provide basic information for the utilization and development of famous classical formulas containing Bletillae Rhizoma, this article systematically analyzes the historical evolution of the name, origin, harvesting and processing of Bletillae Rhizoma by reviewing the ancient materia medica, prescription books, medical books and modern literature. The research results showed that Baiji(白及) was the main name, some scholars took Baiji(白芨) as its main name, and there were many other names such as Baiji(白给), Baigen(白根), Baiji(白苙). The mainstream source of Bletillae Rhizoma was the tubers of Bletilla striata, and drying, large, white, solid, root-free and skin removed completely were the good quality standards. With the promotion of wild to cultivated medicinal materials, there were certain differences between their traits, and the quality evaluation indexes should be adjusted accordingly. The origin of records in the past dynasties was widely distributed, with Guizhou and Sichuan having high production and good quality in modern times. The harvesting period is mostly in spring and autumn, and harvested in autumn was better. The processing and processing technology is relatively simple, and it was used fresh or powdered in past dynasties, while it is mainly sliced for raw use in modern times. Based on the results, it is suggested that the tubers of Bletilla striata of Orchidaceae should be used in the famous classical formulas, and it should be uniformly written as Baiji(白及). And if the original formula indicates the requirement of processing, it should be operated according to the requirement, if the requirement of processing is not indicated, it can be used in raw form as medicine.
2.Protective effect of astaxanthin on acute liver injury induced by α-amanitin in mice
Yongping LUO ; Jiaju ZHONG ; Qunmei YAO ; Zhengxiang GENG ; Chonggui CHEN ; Chengmin YU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(11):801-806
Objective:To explore the protective effect of astaxanthin on acute liver injury induced by α-amanitin in mice.Methods:In June 2023, 42 healthy SPF male Kunming mice were selected. The mice were divided into blank control group, model (0.45 mg/kg α-amanitin) group, olive oil (10 ml/kg olive oil) group, low dose (20 mg/kg) astaxanthin group, medium dose (40 mg/kg) astaxanthin group, high dose (80 mg/kg) astaxanthin group and silybin (20 mg/kg) group by random number table method. Each group had 6 animals. Mice in the blank control group were intraperitoneally injected with 10 ml/kg normal saline, and mice in the other group were injected with α-amanitin. After that, the blank control group and model group were infused with 10 ml/kg normal saline, olive oil group and astaxanthin groups were given olive oil and astaxanthin according to dose by gavage, and silybin group was injected with silybin by dose. The drug was administered once every 12 h for a total of 4 doses. After 60 h, the mice were killed, the liver weight was weighed, and the liver index was calculated. The contents of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum of mice were detected, and the contents of superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT), malondialdehyde (MDA) in liver tissues were also detected. One-way analysis of variance (ANOVA) was used to compare the difference of indexes among each group, and pairwise comparison was performed by Dunnett- t test. Results:The mice in the blank control group had smooth hair color, good spirit and normal behavior, while the mice in the other groups showed varying degrees of retardation and decreased diet, and no death occurred in each group. Body mass[ (26.67±1.51) g] and liver mass[ (1.23±0.14) g] in model group were significantly lower than those in blank control group [ (33.50±2.43) g and (1.87±0.16) g], and the differences were statistically significant ( P<0.05). The liver index [ (5.39±0.32) %, (5.83±0.30) %, (5.75±0.24) % and (5.78±0.16) %] in low, medium and high dose astaxanthin groups and silybin group were significantly higher than those in model group [ (4.61±0.12) %], and the differences were statistically significant ( P<0.05). Serum ALT and AST contents in model group [ (153.04±13.96) U/L and (59.08±4.03) U/L] were significantly higher than those in blank control group [ (13.77±1.29) U/L and (10.21±0.35) U/L], and the differences were statistically significant ( P<0.05). The contents of CAT, GSH and SOD in liver tissues of model group [ (9.40±2.23) U/mgprot, (3.09±0.26) μmol/gprot and (48.94±3.13) U/mgprot] were significantly lower than those of blank control group [ (26.36±2.92) U/mgprot, (6.76±0.71) μmol/gprot and (89.89±4.17) U/mgprot], the differences were statistically significant ( P<0.05). MDA content[ (6.33±0.24) nmol/mgprot] in liver tissue of model group was significantly higher than that of blank control group [ (0.91±0.21) nmol/mgprot], and the difference was statistically significant ( P<0.05). The CAT contents[ (18.64±1.76) U/mgprot, (18.20±1.76) U/mgprot, and (15.54±1.36) U/mgprot] in liver tissues of low, medium and high dose astaxanthin groups were significantly higher than those of model group, with statistical significances ( P<0.05). Compared with model group, SOD contents[ (72.16±7.44) U/mgprot, (93.18±5.28) U/mgprot, (103.78±7.07) U/mgprot, and (96.60±7.02) U/mgprot] in liver tissues of mice in low, medium and high dose astaxanthin groups and silybin group were significantly increased ( P<0.05), MDA contents [ (4.30±0.84) U/mgprot, (3.66±0.28) U/mgprot, (2.96±0.29) U/mgprot, and (2.88±0.39) U/mgprot] were significantly decreased ( P<0.05). Compared with model group, GSH content [ (7.90±1.25) μmol/gprot] in high dose astaxanthin group was significantly increased ( P<0.05) . Conclusion:Astaxanthin may alleviate acute liver injury induced by α-amanitin by alleviating oxidative stress in mice liver.
3.Protective effect of astaxanthin on acute liver injury induced by α-amanitin in mice
Yongping LUO ; Jiaju ZHONG ; Qunmei YAO ; Zhengxiang GENG ; Chonggui CHEN ; Chengmin YU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(11):801-806
Objective:To explore the protective effect of astaxanthin on acute liver injury induced by α-amanitin in mice.Methods:In June 2023, 42 healthy SPF male Kunming mice were selected. The mice were divided into blank control group, model (0.45 mg/kg α-amanitin) group, olive oil (10 ml/kg olive oil) group, low dose (20 mg/kg) astaxanthin group, medium dose (40 mg/kg) astaxanthin group, high dose (80 mg/kg) astaxanthin group and silybin (20 mg/kg) group by random number table method. Each group had 6 animals. Mice in the blank control group were intraperitoneally injected with 10 ml/kg normal saline, and mice in the other group were injected with α-amanitin. After that, the blank control group and model group were infused with 10 ml/kg normal saline, olive oil group and astaxanthin groups were given olive oil and astaxanthin according to dose by gavage, and silybin group was injected with silybin by dose. The drug was administered once every 12 h for a total of 4 doses. After 60 h, the mice were killed, the liver weight was weighed, and the liver index was calculated. The contents of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum of mice were detected, and the contents of superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT), malondialdehyde (MDA) in liver tissues were also detected. One-way analysis of variance (ANOVA) was used to compare the difference of indexes among each group, and pairwise comparison was performed by Dunnett- t test. Results:The mice in the blank control group had smooth hair color, good spirit and normal behavior, while the mice in the other groups showed varying degrees of retardation and decreased diet, and no death occurred in each group. Body mass[ (26.67±1.51) g] and liver mass[ (1.23±0.14) g] in model group were significantly lower than those in blank control group [ (33.50±2.43) g and (1.87±0.16) g], and the differences were statistically significant ( P<0.05). The liver index [ (5.39±0.32) %, (5.83±0.30) %, (5.75±0.24) % and (5.78±0.16) %] in low, medium and high dose astaxanthin groups and silybin group were significantly higher than those in model group [ (4.61±0.12) %], and the differences were statistically significant ( P<0.05). Serum ALT and AST contents in model group [ (153.04±13.96) U/L and (59.08±4.03) U/L] were significantly higher than those in blank control group [ (13.77±1.29) U/L and (10.21±0.35) U/L], and the differences were statistically significant ( P<0.05). The contents of CAT, GSH and SOD in liver tissues of model group [ (9.40±2.23) U/mgprot, (3.09±0.26) μmol/gprot and (48.94±3.13) U/mgprot] were significantly lower than those of blank control group [ (26.36±2.92) U/mgprot, (6.76±0.71) μmol/gprot and (89.89±4.17) U/mgprot], the differences were statistically significant ( P<0.05). MDA content[ (6.33±0.24) nmol/mgprot] in liver tissue of model group was significantly higher than that of blank control group [ (0.91±0.21) nmol/mgprot], and the difference was statistically significant ( P<0.05). The CAT contents[ (18.64±1.76) U/mgprot, (18.20±1.76) U/mgprot, and (15.54±1.36) U/mgprot] in liver tissues of low, medium and high dose astaxanthin groups were significantly higher than those of model group, with statistical significances ( P<0.05). Compared with model group, SOD contents[ (72.16±7.44) U/mgprot, (93.18±5.28) U/mgprot, (103.78±7.07) U/mgprot, and (96.60±7.02) U/mgprot] in liver tissues of mice in low, medium and high dose astaxanthin groups and silybin group were significantly increased ( P<0.05), MDA contents [ (4.30±0.84) U/mgprot, (3.66±0.28) U/mgprot, (2.96±0.29) U/mgprot, and (2.88±0.39) U/mgprot] were significantly decreased ( P<0.05). Compared with model group, GSH content [ (7.90±1.25) μmol/gprot] in high dose astaxanthin group was significantly increased ( P<0.05) . Conclusion:Astaxanthin may alleviate acute liver injury induced by α-amanitin by alleviating oxidative stress in mice liver.
4.Construction of a novel disulfidptosis-related prognostic model for patients with hepatocellular carcinoma based on bioinformatics analysis
Zheng SONG ; Wei LUO ; Xiujuan CHANG ; Yongping YANG
Journal of Clinical Hepatology 2024;40(9):1822-1832
Objective To investigate the expression of disulfidptosis-related genes in hepatocellular carcinoma(HCC)and the prognostic value of disulfidptosis in HCC,to construct a prognostic model,and to analyze its impact on the biological processes of HCC and sorafenib resistance.Methods The TCGA-LIHC database was used to collect the mRNA expression profiles and corresponding clinical data of HCC patients,and the LASSO-Cox regression algorithm was used to construct a four-gene predictive model for prognosis in the TCGA cohort.The external datasets ICGC and GSE14520 were used to validate the prognostic efficacy of the model,and the Cancer Drug Sensitivity Genomics(GDSC)data were used to investigate the value of the disulfidptosis model in predicting sorafenib treatment response,and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were used to investigate the biological functions of disulfidptosis-related genes.The independent-samples t test was used for comparison of continuous data between two groups,and the chi-square test was used for comparison of categorical data between two groups.The Kaplan-Meier curve and the log-rank test were used to evaluate the difference in prognosis,and univariate and multivariate Cox regression analyses were used to investigate whether risk score was an independent influencing factor for patient prognosis.Results The univariate Cox regression analysis in the TCGA cohort showed that seven known disulfidptosis-related genes were significantly associated with overall survival(OS)in HCC(all P<0.05).The LASSO-Cox regression analysis was used to construct a prognostic model based on disulfidptosis-related genes(DRG),and the risk score RS-DRG was calculated as RS-DRG=(0.061 6)×GYS1 expression level+(0.152 8)×LRPPRC expression level+(0.268 3)×RPN1 expression level+(0.183 5)×SLC7A11 expression level.The log-rank test showed that the patients with a high risk score based on the disulfidptosis model had a significantly lower OS than those with a low risk score(P<0.001).Based on the results of the multivariate Cox regression analysis,risk score was an independent predictive factor for OS in both TCGA and ICGC cohorts(TCGA:hazard ratio[HR]=1.869,P=0.002;ICGC:HR=3.469,P=0.004).The Spearman correlation analysis showed that RS-DRG was significantly positively correlated with the infiltration level of various immune cells(including B lymphocytes,CD4+T lymphocytes,neutrophils,macrophages,and dendritic cells)in tumor microenvironment(all P<0.05).The patients in the high-risk score group had a significantly lower IC50 value of sorafenib and were more sensitive to sorafenib(P<0.001).The KEGG/GO enrichment analysis showed that the differentially expressed disulfidptosis-related genes were significantly enriched in various mitosis-related molecular functions.Conclusion This study constructed a novel prognostic model based on disulfidptosis-related genes,which has a potential clinical value in predicting the prognosis of HCC,and targeting disulfidptosis-related genes may provide a promising approach for HCC treatment.
5.Interventional effect of bone marrow mesenchymal stem cell transplantation with different doses of X-ray irradiation induced hepatic injury in mice
Yue LIANG ; Lan LUO ; Tianyu CHENG ; Gaofeng CHEN ; Wei LIU ; Yongping MU ; Jiamei CHEN ; Ping LIU
Chinese Journal of Hepatology 2024;32(11):1019-1027
Objective:To investigate the interventional effect of bone marrow mesenchymal stem cell (BMMSC) transplantation with different doses of X-ray irradiation induced hepatic injury in mice.Methods:Eighteen female C57BL/6J mice were randomly divided into 0, 2, and 3 Gy irradiation groups and 0, 2, and 3 Gy transplantation groups. The irradiation group was used as the control and injected with an equal volume of culture medium. The mice in the transplantation group were irradiated with different doses of X-ray irradiation, and BMMSCs were intravenously infused into the bone marrow. The mice were sacrificed for sampling at the end of the 21st day. Mice body weight changes were recorded daily. The changes in the content of peripheral blood lymphocytes, red blood cells, platelets, and hemoglobin were detected by an automatic blood tester. The morphological changes in mice liver tissues were observed by hematoxylin-eosin staining. The serum activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected by a biochemical analyzer. The reduced glutathione contents in liver tissue were detected by the microplate method. The malondialdehyde content in liver tissue was detected by thiobarbituric acid. The content of total superoxide dismutase (T-SOD) in liver tissue was detected by the hydroxylamine method. The expression of the F4/80 protein in liver tissue was detected by the immunohistochemistry method. The protein expression of nuclear transcription factor erythroid 2 related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) in liver tissue was determined by the western blotting method. The mRNA expression of NLRP3, IL-6, and Nrf2 in liver tissue was detected by a real-time quantitative polymerase chain reaction. The multiple-group comparisons were analyzed by factorial analysis of variance. The inter-group comparisons were analyzed by the LSD method for statistical analysis.Results:The contents of peripheral blood lymphocytes, erythrocytes, platelets, and hemoglobin were significantly decreased in the 3 Gy irradiation group than the 0 Gy irradiation group ( P<0.05), while the activities of serum ALT and AST were significantly increased ( P<0.05). The malondialdehyde content, F4/80 protein expression level, nucleotide-binding domain and leucine-rich repeats, nucleotide oligomerization domain-like receptor family, pyrin domain containing 3 (NLRP3), and interleukin 6 mRNA expression levels were significantly increased in liver tissue, while the contents of T-SOD and glutathione, Nrf2 and HO-1 protein expression levels, and Nrf2 mRNA expression level in liver tissue were significantly decreased ( P<0.05). The contents of peripheral blood lymphocytes, red blood cells, platelets, and hemoglobin were significantly increased in the 3 Gy transplantation group than the 3 Gy irradiation group ( P<0.05), while the activities of serum ALT and AST were significantly decreased ( P<0.05). The malondialdehyde content, F4/80 protein expression level, NLRP3 and interleukin-6 mRNA expression levels in liver tissue were significantly decreased ( P<0.05), while the content of T-SOD and glutathione, Nrf2 and HO-1 protein expression levels, and Nrf2 mRNA expression level in liver tissue were significantly increased ( P<0.05). Conclusion:X-ray irradiation at a dose of 3 Gy can induce liver oxidative damage in mice. BMMSC transplantation can improve X-ray irradiation-induced liver oxidative damage in mice, and its mechanism of action may be related to the regulation of the Nrf2/HO-1 pathway.
6.Association of P-I-R classification and Laennec grading with histology and prognosis after antiviral therapy in patients with hepatitis B cirrhosis
Caihong LYU ; Zheng SONG ; Jing LUO ; Xiujuan CHANG ; Yongping YANG
Journal of Clinical Hepatology 2023;39(3):580-589
Objective To investigate the role of P-I-R classification and Laennec grading in evaluating histological changes in patients with hepatitis B cirrhosis after receiving antiviral therapy, as well as the association of these two evaluation systems with clinical prognosis. Methods A total of 218 patients from 14 centers were consecutively screened from October 2013 to October 2014, and these patients were diagnosed with liver cirrhosis based on pathology (Ishak score ≥5), received antiviral therapy for 72 weeks, completed two liver biopsies, and met the P-I-R classification criteria. The 218 patients were divided into non-hepatocellular carcinoma (HCC) group with 186 patients and HCC group with 32 patients. The chi-square test and the Fisher's exact test were used for comparison of categorical data between groups. For the comparison of HCC after antiviral therapy, the non-parametric Mann-Whitney U test was used for continuous variables, and for the comparison of P-I-R classification and Laennec grading, the non-parametric Kruskal-Wallis H test was used for continuous variables. Univariate and multivariate Cox regression analyses were used to calculate hazard ratio ( HR ) and 95% confidence interval ( CI ), and the Kaplan-Meier method was used to calculate the cumulative incidence rate of HCC. Results After 72 weeks of antiviral therapy, there was a significant difference in P-I-R classification between the non-HCC group and the HCC group ( P < 0.001). There were significant differences in the distribution of Laennec grading and P-I-R classification before and after antiviral therapy ( P < 0.001). After antiviral therapy, the 218 patients were divided into 4A group with 33 patients, 4B group with 71 patients, and 4C group with 114 patients according to Laennec grading, and there were significant differences between these three groups in platelet count (PLT) ( H =36.429, P < 0.001), liver stiffness measurement (LSM) ( H =13.983, P =0.004), Ishak score ( χ 2 =23.060, P < 0.001), and HAI score ( P < 0.001). After antiviral therapy, the 218 patients were divided into R group with 70 patients, I group with 52 patients, and P group with 96 patients according to P-I-R classification, and there were significant differences between these three groups in PLT ( H =7.193, P =0.028), LSM ( H =6.238, P =0.045), Ishak score ( χ 2 =7.986, P < 0.001), HAI score ( P =0.002), and HCC ( P < 0.001). There was a significant difference in the incidence rate of HCC between the P and R groups based on P-I-R classification ( HR =24.21, 95% CI : 0.46-177.99, P =0.002). After adjustment for other confounding factors, P-I-R classification was an independent predictive factor for HCC ( HR =12.69, 95% CI : 4.63-34.80, P =0.002). Conclusion Both P-I-R classification and Laennec grading can reflect the features and changes of fibrosis before and after antiviral therapy, and P-I-R classification is more sensitive to fibrosis changes after antiviral therapy. P-I-R classification (after treatment) can be used to assess the risk of HCC in patients after antiviral therapy.
7.Clinical efficacy and safety of percutaneous cryoablation combined with percutaneous ethanol injection in elderly patients with hepatocellular carcinoma aged 70 years or older
Jing LUO ; Caihong LYU ; Yongping YANG
Journal of Clinical Hepatology 2022;38(2):365-371
Objective To investigate the clinical efficacy and safety of percutaneous cryoablation combined with percutaneous ethanol injection (PEI) in elderly patients with early-stage hepatocellular carcinoma aged 70 years or older. Methods A retrospective analysis was performed for the clinical data of 92 elderly patients with hepatocellular carcinoma who were admitted to The Fifth Medical Center of Chinese PLA General Hospital from January 2014 to January 2018, among whom 46 underwent cryoablation alone (CRYO group) and 46 underwent cryoablation combined with PEI (combination therapy group). The two groups were compared in terms of clinical outcome, adverse reactions, and changes in liver function parameters after treatment, and the patients were followed up to observe tumor recurrence and survival. The t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. The Kaplan-Meier method was used for survival analysis, and the log-rank test was used for comparison of survival curves. The Cox regression analysis was used to identify the independent risk factors for survival and prognosis. Results There was no significant difference in the response rate of initial ablation between the combination therapy group and the CRYO group (89.1% vs 73.9%, P > 0.05). There were no significant differences between the CRYO group and the combination therapy group in overall survival time and tumor-free survival rate after surgery ( P > 0.05), and compared with the CRYO group, the combination therapy group had significantly lower 1-, 2-, and 3-year local tumor progression rates (20%/21%/21% vs 30%/46%/46%, χ 2 =4.187, P < 0.05). The multivariate Cox regression analysis showed that cryoablation alone might be an independent risk factor for local tumor progression( HR =2.206, 95% CI : 1.003-4.850, P =0.049). There was no significant difference in the incidence rate of adverse reactions between the two groups ( P > 0.05), but 3 patients in the CRYO group experienced serious adverse reactions, while no patients in the combination therapy group experienced such reactions. Conclusion For elderly patients with early-stage hepatocellular carcinoma, cryoablation combined with PEI is safer and more effective than cryoablation alone and can significantly reduce local tumor progression rate.
8.Comparison of Improvement Effects and Its Mechanism of Different Doses of Total Alkaloids from Aconitum racemulosum on CIA Model Rats Based on Information Entropy Theory
Xiaomei YANG ; Xiaolan CHEN ; Yongping ZHANG ; Fangfang YANG ; Lang XIE ; Jinghong YANG ; Chengyue LUO ; Hua ZHA
China Pharmacy 2019;30(16):2206-2209
OBJECTIVE: To investigate the effects of different doses of total alkaloids from Aconitum racemulosum (ARTA) on serum inflammation factors and FOS protein expression in synovial tissue of joint in collagen-induced arthritis (CIA) model rats, and to investigate its potential mechanism of anti-rheumatoid arthritis (RA). METHODS: Male SD rats were randomly divided into blank group, model group, positive group (Compound dexamethasone acetate ointment, 0.2 g/kg), ARTA low-dose, medium-dose and high-dose groups (56.26, 112.50, 225.00 mg/kg, by the weight of ARTA in the extract), with 10 rats in each group. Except for blank group, other groups were given subcutaneous injection of Bovine collagen Ⅱ emulsified with incomplete Freund’s adjuvant into the left foot to establish CIA model; the left foot were smeared with relevant medicine from the day of modeling. Blank group and model group were smeared with constant volume of 65% ethanol, 3 times a day, for consecutive 28 days. On the 7th, 14th, 21st and 28th day of administration, the thickness of left hind toe was measured with vernier caliper, and the degree of foot swelling was calculated. The serum contents of IL-1β, IL-6 and TNF-α in rats were measured by ELISA after last administration. The expression of FOS protein in synovial tissue was determined by immunohistochemical method [expressed by HIS]. The comprehensive score was conculated by entropy weight method. Effects of each dosage on above indexes of CIA model rats were evaluated with the comprehensive score. RESULTS: Compared with blank group, the degree of foot swelling, serum content of inflammatory factors and HIS value were increased significantly in model group (P<0.05). Compared with model group, the degree of foot swelling in each administration group, serum contents of IL-1β, IL-6 and TNF-α, HIS in positive group and ARTA high-dose group, serum contents of IL-6 and TNF-α in ARTA medium-dose group as well as serum content of TNF-α in ARTA low-dose group were decreased significantly(P<0.05). Comprehensive score of above indicators were 0.37(positive group), 0.31(ARTA high-dose group), 0.23(ARTA medium-dose group) and 0.09(ARTA low-dose group). CONCLUSIONS: ARTA can improve CIA model rats, and the effect tends to increase with the increase of dose. Above effect may be associated with reducing serum content of inflammatory factors and inhibiting the expression of FOS protein in synovial tissue.
9.Establishment of retroviral vector inducibly expressing RN181 and its expression in hepatocellular carcinoma
Kunping LI ; Suihai WANG ; Xiaozai LUO ; Zhijian LIANG ; Wuye CHEN ; Yongping FANG
Chinese Journal of Hepatobiliary Surgery 2017;23(12):841-844
Objective To study the expression and biological functions of RN181 in SMMC7721 cells,the retroviral vector was constructed.Methods Gene cloning techniques were used to construct pRetrox-TRE3G/RN1S1 recombinant vector.The regulating plasmid pRetroX-TRE3G/RN181 or the response plasmid of pRetroX-Tet3G were respectively cotansfected into GP2-293 cells with Envelope Vector plasmid to package retrovirus after routine identification.Both viruses co-infected target cells SMCC7721,and then were selected by G418 to obtain stable cell lines.The stable cell lines were induced by doxycycline (DOX),and then verified by RT-PCR and Western blotting.CCK-8 was used to evaluate the effect of RN181 on growth of SMMC7721 cells.Results We constructed the recombinant plasmid.Stable recombinant plasmid were verified by screening.And there were significant differences of RN181 between the induced and uninduced cell lines through RT-PCR and Western blot.Conclusions We have successfully constructed the inducible stable RN181 expression SMCC7721 cell,which can be used as an effective cell model to study the biological functions of RN181.We found RN181 could suppress the proliferation and invasion in SMMC7721 cells in vitro.
10.Endoscopic sphincterotomy and laparoscopic cholecystectomy in treatment of gallbladder stones and common bile duct stones in the elderly
Zhijian LIANG ; Yongping FANG ; Kunping LI ; Xiaozai LUO ; Wuye CHEN
Chinese Journal of Hepatobiliary Surgery 2017;23(8):521-525
Objective To study the clinical results of patients with gallbladder stones and common bile duct stones treated either by laparoscopic cholecystectomy followed by endoscopic sphincterotomy (EST + LC) or laparoscopic cholecystectomy + common bile duct exploration (LC + LCBDE) in the elderly patients.Methods A retrospective study was conducted on 96 patients who had common bile duct and gallbladder stones treated from January 2012 to January 2016.The patients were divided into the control group and the observation group.46 patients were in the observation group who underwent LC three days after EST,while the remaining patients were in the control group who underwent LC and LCBDE.The serum amylase levels before LC,operation time,intraoperation bleeding volume,postoperative time to first flatus,total hospitalization stay and total hospitalization costs and incidences of postoperative complication were compared.Results The total operation time was (95.0 ±7.0) minutes and (125.0 ± 18.0) minutes,respectively,(P<0.05).The total costs in the two group were (39515.0 ±4 135.0) yuan and (28287.0 ± 2 254.0) yuan (P < 0.05),respectively.Postoperative complications were observed in 5 (6.1%) and 10 (13.2%) patients (P < 0.05),respectively.The preoperative serum amylase levels were (97.6 ± 48.5) IU/L and (131.4 ± 68.7) IU/L,respectively.The blood loss was (35.7 ± 8.5) ml and (31.8 ± 7.3) ml,respectively.The postoperative time to first flatus was (1.7 ± 0.5) days and (1.9 ± 0.4) days,respectively.The total hospitalization stay was (16.3 ±2.8) days and (15.2 ±3.7) days.There were no significantly differences (P > 0.05).Conclusions LC carried out on day 3 after EST to treat elderly patients with cholecystolithiasis and choledocholithiasis was safe and efficacious and the treatment had the advantages of minimal trauma,short operative time,rapid recovery and low complication rates.This should be recommended in clinical practice.

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