OBJECTIVE To explore the ameliorative effect of ursolic acid on carbon tetrachloride-induced acute liver injury in mice,and the feasibility of multispectral optoacoustic tomography(MSOT)for characteristic structural and functional imaging of liver tissues.METHODS Kunming mice were randomly divided into the normal control group,model group,model+ursolic acid 30,60 mg·kg-1 groups and model+bifendate 5.625 mg·kg-1 group,with 14 mice in each.Each group was given the corresponding drug once daily for 7 days.An acute liver injury model was established in mice by intraperitoneal injection of 0.2％carbon tetrachloride in olive oil solution after the last administration.Blood was collected,liver tissues were taken 24 h after modeling,and the liver index was calculated,8 mice from each group and the levels of serum glutamic pyruciv transaminase(GPT)and glutamic oxaloacetic transaminase(GOT),as well as superoxide dismutase(SOD)activity and malondialdehyde(MDA)content in liver tissues were measured.The enzyme-linked immunosorbent assay(ELISA)method was used to detect the level ofα-glutathione S-transferase(α-GST)in serum.The histopathological changes of the liver were observed under a light microscope.The remaining 6 mice in each group underwent MSOT technique was used for characteristic structural and functional imaging of liver tissue.Levels of oxygenated hemoglobin(HbO2)and deoxygenated hemoglobin(Hb)were analyzed,oxygen saturation was calculated,and the extent of liver injury was assessed by examining the intrahepatic distribution of indocyanine green(ICG).RESULTS Compared with the normal group,the levels of GPT,GOT and α-GST in serum,content of MDA in liver tissues and the liver index in the model control group were significantly increased while the activity of SOD in liver tissues were significantly decreased.Compared with the model group,ursolic acid in each dose group significantly reduced the liver index of mice,lowered the serum levels of GPT and GOT as well as the level of α-GST,decreased the content of MDA in liver tissues,and elevated the activity of SOD in liver-injured mice.Hematoxylin-eosin staining showed that significant steatosis and hepatocyte necrosis and inflammatory cell infiltration in hepatocytes of mice in the model group.Ursolic acid significantly attenuated the degree of hepatocellular lesions and markedly reduced steatosis in mice.MSOT imaging showed that the HbO2 level and oxygen saturation were significantly lower while the Hb level was remarkably higher in the liver of mice in the model group.In each administration group,the level of HbO2 significantly increased,the level of Hb was significantly decreased,oxygen saturation was significantly increased in the liver of model mice and the accumulation of ICG dye probe was atten-uated in the body after hepatocyte injury.CONCLUSION Ursolic acid can elevate the hepatic oxygen saturation,improve the metabolism of ICG,reduce the degree of hepatic necrosis in mice,and help protect against carbon tetrachloride-induced hepatic injury in mice.The mechanism is probably related to the inhibition of oxidative stress.

OBJECTIVE To explore the ameliorative effect of ursolic acid on carbon tetrachloride-induced acute liver injury in mice,and the feasibility of multispectral optoacoustic tomography(MSOT)for characteristic structural and functional imaging of liver tissues.METHODS Kunming mice were randomly divided into the normal control group,model group,model+ursolic acid 30,60 mg·kg-1 groups and model+bifendate 5.625 mg·kg-1 group,with 14 mice in each.Each group was given the corresponding drug once daily for 7 days.An acute liver injury model was established in mice by intraperitoneal injection of 0.2％carbon tetrachloride in olive oil solution after the last administration.Blood was collected,liver tissues were taken 24 h after modeling,and the liver index was calculated,8 mice from each group and the levels of serum glutamic pyruciv transaminase(GPT)and glutamic oxaloacetic transaminase(GOT),as well as superoxide dismutase(SOD)activity and malondialdehyde(MDA)content in liver tissues were measured.The enzyme-linked immunosorbent assay(ELISA)method was used to detect the level ofα-glutathione S-transferase(α-GST)in serum.The histopathological changes of the liver were observed under a light microscope.The remaining 6 mice in each group underwent MSOT technique was used for characteristic structural and functional imaging of liver tissue.Levels of oxygenated hemoglobin(HbO2)and deoxygenated hemoglobin(Hb)were analyzed,oxygen saturation was calculated,and the extent of liver injury was assessed by examining the intrahepatic distribution of indocyanine green(ICG).RESULTS Compared with the normal group,the levels of GPT,GOT and α-GST in serum,content of MDA in liver tissues and the liver index in the model control group were significantly increased while the activity of SOD in liver tissues were significantly decreased.Compared with the model group,ursolic acid in each dose group significantly reduced the liver index of mice,lowered the serum levels of GPT and GOT as well as the level of α-GST,decreased the content of MDA in liver tissues,and elevated the activity of SOD in liver-injured mice.Hematoxylin-eosin staining showed that significant steatosis and hepatocyte necrosis and inflammatory cell infiltration in hepatocytes of mice in the model group.Ursolic acid significantly attenuated the degree of hepatocellular lesions and markedly reduced steatosis in mice.MSOT imaging showed that the HbO2 level and oxygen saturation were significantly lower while the Hb level was remarkably higher in the liver of mice in the model group.In each administration group,the level of HbO2 significantly increased,the level of Hb was significantly decreased,oxygen saturation was significantly increased in the liver of model mice and the accumulation of ICG dye probe was atten-uated in the body after hepatocyte injury.CONCLUSION Ursolic acid can elevate the hepatic oxygen saturation,improve the metabolism of ICG,reduce the degree of hepatic necrosis in mice,and help protect against carbon tetrachloride-induced hepatic injury in mice.The mechanism is probably related to the inhibition of oxidative stress.

ObjectiveTo statistically analyze the efficacy and application characteristics of animal-derived drugs in the 2020 edition of the Pharmacopoeia of the People's Republic of China （referred to as Chinese Pharmacopoeia）， and provide data references for the research and application of animal-derived drugs. MethodAnimal-derived drugs included in the 2020 edition of the Chinese Pharmacopoeia were selected and screened. Information such as the classification， nature and flavor， meridian tropism， usage， dosage， toxicity or contraindications of single animal-derived drugs were statistically analyzed. The efficacy and clinical application analysis of externally applicable Chinese medicinal preparations containing animal-derived drugs， the dosage forms of preparations containing animal-derived drugs， and the contraindications of their use were also analyzed. The above information was recorded in an Excel spreadsheet for statistical analysis. ResultThe 2020 edition of the Chinese Pharmacopoeia clearly records a total of 51 types of animal-derived drugs. Among them， liver-calming and wind-relieving drugs and deficiency-tonifying drugs are the most common in the efficacy classification of animal-derived drugs. The properties are mainly categorized as neutral and salty， with meridians including liver， kidney， heart， and lung meridians with a frequency of >10 times. Most animal-derived drugs are used in decoctions and pills， with the decoction method being the primary method for decoction. The common dosage of animal-derived drugs is 3-9 g， 5-10 g， or 3-6 g， and animal-derived drugs are used externally in appropriate amounts. In addition to mentioning the toxicity of drugs， the most common precautions for animal-derived drugs are caution and prohibition for pregnant women. Externally applicable Chinese medicinal preparations containing animal-derived in the Chinese Pharmacopoeia are widely used in various departments such as pediatrics， gynecology， orthopedics and traumatology， stomatology， internal medicine， surgery， dermatology， ophthalmology， and otorhinolaryngology， with extensive clinical applications. The dosage forms of preparations containing animal-derived drugs are mainly pills， capsules， tablets， and capsules. Animal-derived drugs are mainly in the form of fine powder before use. The precautions for use in preparations mainly focus on the contraindications for pregnant women and children. ConclusionThe clinical application of animal-derived drugs included in the 2020 edition of Chinese Pharmacopoeia is extensive and has unique efficacy. However， there are still problems such as imperfect quality standards and unclear mechanisms of action for animal-derived drugs， which require further research to better utilize the advantages of internal and external use of animal-derived drugs. In addition， non-medicinal parts of animals also have development and utilization value， which should be fully explored and utilized to achieve the green development of animal-derive drug resources.

Objective To explore the correlation between ATF5 protein expression level and drug sensitivity of gastric cancer AGS cells.Methods We established the high,middle and low ATF5 expression groups by transfecting the AGS cells with ATF5 pcDNA3.1 plasmid,ATF5 siRNA and empty plasmid using the liposome transfection technique.Then we detected the expression of the ATF5 protein of the 3 groups using the Western Blot.The drug sensitivity of AGS cells in 3 groups to paclitaxel and cisplatin was detected using the MTT experiments and plate clone formation experiments then the correlation between ATF5 protein expression level and drug sensitivity of gastric cancer AGS cells was analyzed.Results With the effect of paclitaxel or cisplatin on AGS cells in 3 groups,the surviving fraction and the IC50 of high ATF5 expression group were higher than those of control group,which indicated statistical significance (all P ＜ 0.05);while the surviving fraction and the IC50 of low ATF5 expression group were lower than those of control group (all P ＜ 0.05,P cisplatin =0.00,P paclitaxel =0.002).We found that cell clone formation of AGS cells to paclitaxel and cisplatin was significantly increased after the transfection with ATF5 protein plasmid and decreased after transfection with ATF5 siRNA plasmid (all P ＜ 0.05).Conclusion The expression of ATF5 protein is related to the drug sensitivity of gastric cancer AGS cells that upregulation of ATF5 protein expression can decrease the drug sensitivity of AGS cells to paclitaxel and cisplatin in gastric cancer.

Objective To investigate the effects of silencing STMN1 by siRNA on the sensitivity of oesophageal cancer cells Eca-109 to paclitaxel .Methods The STMN1 siRNA(siSTMN1) or scramble siRNA(SCR) were transient transfected into Eca-109 cells .The mRNA and protein levels of STMN1 were detected by qPCR and Western blot in the Eca-109 cells of different groups .In vitro paclitaxel sensitivity of siSTMN1 and SCR transfected Eca-109 cell lines was tested by MTT assay and colony formation as-say .Hoechst 33258 nuclear staining were used to investigate the effect of silencing STMN 1 on the sensitivity of SCR ,siSTMN1 transfected Eca-109 cells and nontreated counterparts under paclitaxel induced apoptosis .Results The transient transfection cell lines were successfully established .Both protein and mRNA levels of STMN1 were effectively down-regulated in the siSTMN1 transfected Eca-109 cells .Down-regulation of STMN1 significantly enhanced the sensitivity of Eca-109 cells in response to paclitaxel (P<0 .01) .In addition ,the siSTMN1 transfected Eca-109 cells displayed significant apoptosis as assessed by Hoechst nuclear stai-ning(P<0 .01) .Conclusion Silencing STMN1 by siRNA could enhance the sensitivity of oesophageal cancer cells Eca-109 to paclitaxel .

Objective To evaluate the significance of electronic gastroscopy examination combined with miR-21 in the diagnosis of gastric carcinoma.Methods 96 gastroscopy cases in Guangzhou General Hospital of Guangzhou Military Command were adopted from June,2011 to December,2012.The patients received gastric endoscopy examination,and the diseased tissue samples were taken out at the same time.To calculate the diagnosis rate and missed rate with gastroscopy,by comparing the gastroscopy diagnosis with the pathology diagnosis.To evaluate the expression level of miR-21 of the tissue by applying quantitative real time PCR.Results The diagnosis rates of gastric cancer,gastric ulcer,chronic superficial gastritis,gastroesophageal reflux gastritis,chronic atrophic gastritis under gastroscopy were 28.12 ％ (27/96),30.21％ (29/96),18.75 ％ (18/96),10.42 ％ (10/96),12.50 ％ (12/96).The diagnosis rate of gastric cancer was 100.00 ％ and the missed diagnosis rate was 6.90 ％ (2/29),the relative transcript level of miR-21 in the gastric cancer group was 54.41±6.66,which was obviously higher than the gastric ulcer group and the other groups (P ＜ 0.01).Conclusions Electronic gastroscopy is a clinically practical examination method in screening gastric cancer cases,evaluating the expression level of miR-21 in the diseased tissue under gastroscopy can enhance the accuracy of screening gastric cancer.Combining the two methods is useful in the diagnosis of gastric cancer.

Objective To evaluate the clinical effects of early endoscopic therapy for elderly patients with acute severe biliary pancreatitis.Methods Ninety-two elderly patients with acute severe biliary pancreatitis were randomly divided into 2 groups:ERCP group ( n =43) and non-ERCP group ( n =49).Serum TNF-a,IL-6,IL-8,the recovery time of blood amylase,the duration of abdominal pain,hospitalization,mortality and complications were compared.Results ERCP group showed a greater decrease in serum TNF-α,IL-6 and IL-8 levels than the control group ( 45.16 ± 13.48 ) μg/L v.s.( 176.89 ± 47.35 ) μg/L,(31.76 ± 13.85)μg/L v.s.(68.48 ±24.87) μg/L,(113.39 ±63.78) μg/L v.s.(309.86 ± 117.13)μg/L) (P ＜0.05 ).The duration of abdominal pain,the recovery time of blood amylase and hospitalization in ERCP group were significantly shorter compared to the non-ERCP group [ ( 10.2 ± 1.7 ) d v.s.( 13.2 ±2.4)d,(3.3 ±1.0)dv.s.(5.5 ±1.2)d,(15 ±1.6)dv.s.(20±3.0)d] (P＜0.05),and complication rate of the ERCP group was lower,too (5％ v.s.22％,P ＜ 0.05).Conclusion Early ERCP is safe and highly effective for the elderly patients with acute severe biliary pancreatitis.

Objective To investigate the regulation of metabolize characteristic of some antibiotics in ACST in human bile, and provide the theoretic basis for physician selecting antibiotic rationally when biliary tract has been infected. Methods Samples of ACST in human bile were obtained by ERCP + ENBD, contents of Cefradine, Cefoperazone, Ceftriaxone, Ciprofloxacin and Lomefloxacin in human bile were simultaneously measured by HPLC method. Results After antibiotics were given through iv 0. 25 ～ 1. 5 hour, an increase was observed in contents of all antibiotics in bile, and the changes of Ceftriaxone and Ciprofloxa-cin were the most significant ( P <0.01). Ceftriaxone and Ciprofloxacin had a higher Cmax than other antibiotics, and Cefoperazone had a longer tl/2. Conclusion As maintaining a little time to tmax , a higher Cmax and a longer tl/2 in the bile by iv, Cefradine and Ciprofloxacin are recommended in the patient with ACST.

【Objective】 To explore a method for the construction of antitumor suicide-gene therapeutic system. 【Methods】 The thymidine kinase gene of herpes simplex virus type 1 (HSV1-tk) was orientationally cloned into the retroviral vector plasmid (pLXSN) by DNA recombinant technique, and then the recombinant plasmid of pLXSN-tk was identified by restriction endonuclease cutting and DNA sequencing. Introduced by PolyFect Transfection reagent, the recombinant plasmid was transfected into the packaging cell line PT67. Screened by G418, a cell line, which could produce virus stably, was obtained. The virus was transfected into human gastric carcinomatous cell strain SGC-7901 and the transfected SGC-7901 was applied to evaluate an anti-tumor effect. 【Results】 The identification with restriction endonuclease cutting and DNA sequencing showed that HSV1-tk was successfully inserted into the recombined plasmid of pLXSN-tk. The stable cloned PT67/tk was obtained by screening with G418 and then its amount was enlarged by culture, the titer of the PT67/tk solution being 4?10~4 cfu/mL. The anti-G418 cloned cell line SGC-7901/tk was got by infecting SGC-7901 with the virus. Anti-tumor experiment showed that GCV had an obvious toxic effect on SGC-7901/tk but had no effect on SGC-7901, indicating recombinant retroviral HSV1-tk expressed HSV1-tk gene which had biological activity. 【Conclusion】 Cloning HSV1-tk into the retroviral vector is effective in obtaining the recombinant retroviral HSV1-tk which can express HSV1-tk gene, and also effective in establishing an antitumor suicide-gene therapeutic system. This research naturally lays a foundation for further studying of Chinese medicines having synergistic anti-tumor action with suicide gene.

Objectives:To observe growth suppressive effects of human wild type p53(wt p53) gene on human esophageal cancer cell line. Methods:Using the retroviral vector to introduce exogenous wt p53 gene into human esophageal cancer cell line ECA109,the gene expression and tumor inhibition were studied in vitro and in vivo . Results:The expression of p53 in transfected cell lines(ECA 109/p53 ) was increased.The growth rates and the ability to form colony in soft agar were greatly inhibited in ECA 109/p53 cells versus ECA109/neo and ECA109 cells.The G 0+G 1 ratio increased and S ratio decreased in cell cycle distribution,and apoptosis index significantly rose in the ECA109/p53 cells,which were confirmed by FCM analyzing.The tumorigen icity of ECA109/p53 cells in nude mice was obviously suppressed by exogenous p53. Conclusions:Exogenous wt p53 mediated by retroviral vector could inhibit the growth of human esophageal cancer cell.