Role and mechanism of action of phytoestrogen biochanin A in improving liver fibrosis in ovariectomized mice / 临床肝胆病杂志

ObjectiveTo investigate the effect of phytoestrogen biochanin A （BCA） on liver fibrosis induced by CCl4 in female mice with bilateral oophorectomy （ovariectomized） and its mechanism. MethodsA total of 50 ovariectomized Kunming mice were selected and given intraperitoneal injection of CCl4 to establish a model of liver fibrosis， and then according to body weight， they were randomly divided into model group， positive control group， and low-， middle-， and high-dose BCA groups， with 10 mice in each group. In addition， 10 female mice in the same litter were given resection of a small amount of adipose tissue near both ovaries to establish the sham-operation group. The mice in the positive control group were given estradiol 2 mg/kg by gavage， and those in the low-， middle-， and high-dose BCA groups were given BCA by gavage at a dose of 25， 50， and 100 mg/kg， respectively， once a day for 7 consecutive weeks； the mice in the sham-operation group and the model group were given an equal volume of 0.5% sodium carboxymethyl cellulose solution by gavage. The mice were anesthetized and sacrificed after administration to collect samples. Liver index and uterus index were measured； HE staining and Masson staining were used to observe liver histopathological changes； the biochemical analysis was used to measure the activity of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）； ELISA was used to measure the levels of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in liver tissue， and Western blot was used to measure the relative protein expression levels of collagen Ⅰ， transforming growth factor-beta 1 （TGF-β1）， alpha-smooth muscle actin （α-SMA）， estrogen receptor beta （ERβ）， and p-NF-κBp65/NF-κBp65 in liver tissue. The t-test was used for comparison of continuous data between two groups； a one-way analysis of various was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and further comparison between two groups. ResultsCompared with the sham-operated group， the model group had a significant increase in liver index and a significant reduction in uterus index， as well as significant increases in the activities of serum AST and ALT， the levels of IL-6 and TNF-α in liver tissue， and the protein expression levels of collagen Ⅰ， TGF-β1， α-SMA， and p-NF-κBp65/NF-κBp65 in liver tissue （all P<0.05）， with no significant change in the expression of ERβ in liver tissue （P>0.05）， and the model group showed significant fibrosis lesions in the liver， such as hepatocyte edema， steatosis， and necrosis with inflammatory cell infiltration and hyperplasia， deposition， and staggered distribution of collagen fibers. Compared with the model group， the low-， middle-， and high-dose BCA groups had significant reductions in liver index， the activities of serum AST and ALT， the levels of IL-6 and TNF-α， and the protein expression levels of collagen Ⅰ， TGF-β1， α-SMA， and p-NF-κBp65/NF-κBp65 in liver tissue （all P<0.05）， with no significant change in uterine index （P>0.05）， as well as a significant increase in the protein expression level of ERβ in liver tissue （P<0.05） and varying degrees of improvement in liver fibrosis lesions. ConclusionBCA can effectively improve CCL4-induced liver fibrosis in ovariectomized female mice， possibly by upregulating ERβ to inhibit the NF-κB signaling pathway and then alleviating inflammatory response.

Protective effects of twin drug ST-11 against PC12 cell injury induced by oxygen-glucose deprivation/reperfusion and its mechanism / 中国药房

OBJECTIVE To study the protective effects of ligustrazine-scutellarein twin drug ST-11 on rat adrenal medullary pheochromocytoma PC12 cell injury induced by oxygen-glucose deprivation/reperfusion （OGD/R） and its mechanism. METHODS PC12 cells were divided into blank group， model group， nimodipine group （positive control， 5 μmol/L） and different concentration groups of ST-11 （5， 10， 20 μmol/L）. After 24 hours of pre-administration intervention， all the other groups except the blank group were cultured in glucose-free DMEM culture medium containing 10 mmol/L Na2S2O4 for 4 hours with glucose deficiency and hypoxia. After 4 hours of glucose and oxygen re-introduction， the survival rate of cells in each group， the contents of lactate dehydrogenase （LDH）， catalase （CAT）， glutathione （GSH）， malondialdehyde （MDA） and superoxide dismutase （SOD） in cell supernatant， apoptosis rate， the levels of reactive oxygen species （ROS） and mitochondrial membrane potential （MMP）， the protein expressions of B-cell lymphoma 2 related X protein （Bax）， B-cell lymphoma 2 （Bcl-2） and caspase-3 were all detected in each group. RESULTS Compared with blank group， the cell survival rate， the contents of CAT， GSH and SOD in cell supernatant， MMP level， relative expression of Bcl-2 and Bcl-2/Bax ratio in model group decreased significantly （P＜0.05）， while the contents of LDH and MDA， ROS level， apoptosis rate， relative expressions of Bax and caspase-3 were significantly increased （P＜0.05）. Compared with model group， above indexes of ST-11 groups （except for the protein expression of caspase-3 in 5 μmol/L ST-11 group） were reversed signifi-cantly （P＜0.05）. CONCLUSIONS ST-11 has a certain protec-tive effect on OGD/R-injured PC12 cells， and its effects may be related to reduction of oxidative stress and inhibition of cell apoptosis.

Effects and mechanism of the ethanol extract of Tiarella polyphylla on learning and memory impairment in mice / 中国药房

OBJECTIV E To investigate the effect s and mechanism of the ethanol extract of Tiarella polyphylla （“TPE”）on learning and memory impairment in mice. METHODS Male Kunming mice were randomly divided into normal group ，model group，positive group （donepezil hydrochloride 4 mg/kg）and TPE low-dose ，medium-dose and high-dose groups （150，300，600 mg/kg），with 10 mice in each group. Drug administration groups were given relevant medicine intragastrically once a day ，and normal group and model group were given water intragastrically once a day ，for consecutive 22 d. On the 17th day ，administration groups and model group were intraperitoneally injected with scopolamine hydrobromide （3 mg/kg）to establish a model of learning and memory impairment. The learning and memory ability of the mice were evaluated by the Morris water maze. Hematoxylin-eosin （HE） staining was used for morphological observation of hippocampus cells of the mice. The levels of acetylcholinesterase （AChE），choline acetyltransferase （ChAT），superoxide dismutase （SOD），malondialdehyde（MDA），interleukin-6（IL-6）and tumor necrosis factor-α（TNF-α）in cerebral tissue as well as the relative expression of phosphorylated Tau protein （p-Tau），β-site amyloid precursor protein cleaving enzyme 1（BACE1）and amyloid precursor protein （APP）in hippocampus tissue were all detected. RESULTS The escape latency of mice in positive group ，TPE medium-dose and high-dose groups were all significantly shortened than the model group on the 4th to 5th day of training ，while the times of crossing platform and the percentage of movement distance in target quadrant were significantly increased （P＜0.05）. Compared with model group ，the neurons in the hippocampal CA 1 region of mice were increased to var ying degrees in administration groups ，the ne urons in solidified and atrophic state decreased ，and the arrangement of neurons tended to be close；the levels of ChAT and SOD in cerebral tissue were significantly increased in positive group and TPE medium-dose and high-dose groups ；the levels of AChE ，MDA，IL-6，the levels of TNF-α and relative expression of p-Tau ，BACE1 and APP in hippocampus tissue were decreased significantly （P＜ 0.05）. CONCLUSIONS TPE can improve the learning and memory impairment induced by scopolamine in mice ，and the mechanism may be related to balancing the brain cholinergic system ，alleviating oxidative stress injury ，improving inflammatory response，and inhibiting the overexpression of p-Tau ，BACE1 and APP .

Effects of water extract and ethanol extract from the root of Caragana sinica on hyperuricemia in mice / 中国药房

OBJECTIVE To investigat e the effects of water extract （WCS）and ethanol extract （ECS）from the root of Caragana sinica on hyperuricemia （HUA）in mice. METHODS Kunming mice were randomly divided into normal control group ， model group ，allopurinol group （positive control ，5 mg/kg），benzbromarone group （positive control ，7.8 mg/kg），WCS low-dose ， medium-dose and high-dose groups （38，75，150 mg/kg），ECS low-dose ，medium-dose and high-dose groups （50，100，200 mg/kg）， with 10 mice in each group. Except for the normal control group ，the other mice were given potassium oxazinate intraperitoneally and hypoxanthine intragastrically for consecutive 7 d to establish HUA model. On the third day of modeling ，mice in each administration group were given corresponding drugs intragastrically ，and normal control group and model group were given equal volume of normal saline once a day for 5 consecutive days.The body weight of mice were weighted during administration ；one hour after the last administration ，the organ indexes of liver ，kidney and spleen were calculated ；the contents of serum uric acid （SUA）， blood urea nitrogen （BUN）and serum creatinine （SCR）；the activity of xanthine oxidase （XOD）in serum and liver tissue were determined. Relative mRNA and protein expressions of XOD in liver tissue ，relative expre ssions of GLUT9，URAT1 and OAT 1 in renal tissue were all detected ；and the pathological changes of renal tissue were observed. RESULTS There were no significant differences in liver index and spleen index in each group （P＞0.05）. Compared with normal control group ， except for allopurinol group ， there were no significant differences in the body weight and the contents of BUN and SCR in mice of other administration groups （P＞0.05）；the renal index and SUA content of mice in the m odel group and allopurinol group were significantly increased （P＜0.05）；in the model group ，the XOD activity in serum and liver tissue ，the relative mRNA and protein expression of XOD in liver tissue ，the relative expressions of GLUT 9 and URAT 1 protein in renal tissue were significantly increased （P＜0.05），and the relative expression of OAT 1 protein in renal tissue was significantly decreased （P＜ 0.05）. Compared with model group ，renal indexes of mice were decreased significantly in WCS and ECS groups （P＜0.05），and the pathological damage of renal tissue was significantly improved ；SUA content ，XOD activity in serum and liver tissue ，the relative mRNA and protein expression of XOD in liver tissue ，and the relative expression of URAT 1 protein in renal tissue were decreased significantly in administration groups （P＜0.05）. The relative expression of GLUT 9 protein in renal tissue of mice in benzbromarone group and ECS high-dose group decreased significantly （P＜0.05）；relative expression of OAT 1 protein in renal tissue of mice in benzbromarone group ，WCS low-dose and high-dose groups ，ECS low-dose group were increased significantly （P＜0.05）. CONCLUSIONS WCS and ECS can significantly decrease the contents of SUA in HUA model mice ，and improve pathological state of renal tissue ，the mechanism of which may be associated with inhibiting XOD activity and uric acid reabsorption，and down-regulating protein and mRNA expression of XOD.

Preliminary Study on Improvement Effect of Tiarella polyphylla Ethanol Extract on CCl 4-induced Hepatic Fibrosis in Mice and Its Mechanism / 中国药房

OBJECTIVE：To investigate the improvement effect of Tiarella polyphylla ethanol extract （TPME）on CCl 4-induced hepatic fibrosis in mice ，and to explore its possible mechanism preliminarily. METHODS ：Totally 60 male Kunming mice were randomly divided into normal group ，model group ，positive control group （colchicine 0.1 mg/kg），TPME low-dose ，medium-dose and high-dose groups （250，500，1 000 mg/kg）according to body weight ，with 10 mice in each group. Except for normal group ， other groups were given 20％ CCl4 olive oil solution intraperitoneally to induce hepatic fibrosis ，twice a week ，for consecutive 8 weeks. From the fifth week after modeling ，administration groups were given relevant medicine intragastrically ，normal group and model group were given constant volume of normal saline intragastrically ，once a day ，for consecutive 4 weeks. Twelve hours after last administration ，the liver weight of mice in each group was measured and the liver index was calculated. The serum contents of ALT，AST，SOD，MDA，PC-Ⅲ，C-Ⅳ，LN，TNF-α and IL- 6 were determined. Western blot assay was used to detect the protein expression of α-SMA，TGF-β1 and Smad 3 in liver tissue. HE and Masson staining were used to observe the pathological changes of hepatic tissue. RESULTS ：Compared with normal group ，the liver index ，the activities of ALT and AST and the contents of MDA ， LN，PC- Ⅲ ，C- Ⅳ ，LN，TNF-α and IL- 6 in serum were increased significantly ， while the activity of SOD was 6011） decreased significantly in model group （P＜0.01）；the protein expression of α-SMA，TGF-β1 and Smad 3 in liver tissues were hfjsznd8@126.com increased significantly （P＜0.01）. Obvious fibrosis lesions was observed in liver tissue. Compared with model group ，the live indexes ，the activities of ALT and AST ，the contents of MDA，PC-Ⅲ，C-Ⅳ，LN，TNF-α and IL-6 in serum were decreased significantly in positive control group and TPME groups ， while the activities of SOD were increased significantly （P＜0.05 or P＜0.01）. The protein expression of α-SMA，TGF-β1 and Smad3 in liver tissue were decreased significantly （P＜0.05 or P＜0.01），and liver fibrosis was improved to different extent. Compared with TPME low-dose group ，the contents of PC- Ⅲ，LN and IL- 6 in serum ，protein expression of TGF-β1 and Smad 3 in liver tissue were decreased significantly in TPME high-dose group （P＜0.05）. CONCLUSIONS ：TPME can improve hepatic fibrosis induced by CCl 4 in mice ，the mechanism of which may be associated with the inhibition of collagen synthesis and oxidative stress，the reduction of inflammatory factors ，and the down-regulation of the expression α-SMA and relative proteins of TGF-β1/ Smad signal pathway.

Clinical application progress of variations in the technique of liver partition from ALPPS / 国际肿瘤学杂志

There are many technical variations and improvements in associating liver partition and portal vein ligation for staged hepatectomy (ALPPS). Its main purpose is to reduce the incidence of complications and mortality, and at the same time, the future liver remnant compensation can be increased more faster and its function becomes more robust. In clinical practice, different researchers have studied the variation and related clinical results of liver disconnection technique in ALPPS, such as partial hepatectomy, portal vein ligation assisted radio frequency hepatectomy, laparoscopic microwave ablation and portal vein ligation staged hepatectomy, combined liver tourniquet and portal vein ligation staged hepatectomy, continuous combined liver tourniquet and portal vein ligation staged hepatectomy, in situ hepatectomy combined with portal vein embolization via anterior approach and portal vein embolization instead of hepatectomy separation of visceral substance, and so on.

Effects of doxorubicin-loaded tumor-derived extracellular vesicles on cell proliferation and apoptosis of human hepatocellular carcinoma / 中华肝胆外科杂志

Objective:To investigate the effects of doxorubicin (Dox)-loaded tumor-derived extracellular vehicles (EVs) on cell proliferation and apoptosis of human hepatocellular carcinoma.Methods:The extracellular vesicles loaded with Adriamycin (EVs-Dox) were prepared by the method of directly co-incubation. The morphology of EVs-Dox was detected by transmission electron microphotometer. The diameter of EVs-Dox was determined by dynamic light scattering (DLS). Western blotting was utilized to detect the expression of CD63, HSP 70 and TSG 101 in the EVs-Dox. The encapsulation efficiency of EVs-Dox was calculated by tandem mass spectrometry (LC-MS/MS). The drug release experiment in vitro was utilized to simulate the drug release of drug-loaded vesicles in vivo. PKH67-labeled EVs-Dox was showed cellular uptake. After treatment with EVs-Dox, MTS assay and flow cytometry assay were conducted to investigate the effects of EVs-Dox on cell proliferation and apoptosis of PLC/PRF/5.Results:The EVs-Dox showed an elliptical double-layer membrane structure of different sizes under transmission electron microscope. The diameter of EVs-Dox was (115.9±5.2) nm.Western blotting data showed high expression of CD 63, HSP 70 and TSG 101 in the EVs-Dox. The encapsulation efficiency of EVs-Dox was 0.77%. The in vitro release experiment showed that the drug-loaded vesicles could release the drug slowly. PKH67-labeled EVs-Dox showed that carcinoma cells can uptake EVs-Dox within 16h. MTS assay showed that the cell viability rate of (54.9±3.2) % was significantly lower than that of in the Dox group [(77.7±5.4)%, P<0.05]. EVs-Dox inhibited hepatocellular carcinoma proliferation. Flow cytometry assay showed that the apoptosis rate of EVs-Dox (47.9±7.0) % was higher than that in the Dox group [(38.0±1.5)%, P<0.05]. Conclusion:EVs-Dox inhibits cell proliferation and accelerates apoptosis of hepatocellular carcinoma cells.

Inhibitory Effects of Protopine on the Proliferation of Human Hepatic Stellate Cells HSC-LX 2 and Its Mechanism Study / 中国药房

OBJECTIVE： To investigate inhibitory effects of protopine on the proliferation of human hepatic stellate cells HSC-LX2 and to explore its mechanism preliminarily. METHODS： MTT assay was used to detect the effects of 25， 50， 100， 200， 400 and 500 μmol/L protopine on the proliferation of HSC-LX2 cells. The inhibitory effect of cell proliferation was calculated. HSC-LX2 cells were divided into control group （1640 medium containing 5％ fetal bovine serum）， protopine low-concentration， medium-concentration and high-concentration groups （100， 200， 400 μmol/L）. After treated for 24 h. The apoptotic rate of the cells was detected by flow cytometry. RT-PCR was used to determine the mRNA expression of α-SMA， Collagen Ⅰ， Collagen Ⅲ， MMP-2 and TIMP-1 in cells. The protein expressions of α-SMA， Collagen Ⅰ， Collagen Ⅲ and MMP-2 were detected by Western blot. RESULTS： The inhibitory rates of 25， 50， 100， 200， 400 and 500 μmol/L protopine on proliferation HSC-LX2 cells were 0， 6.9％， 18.7％， 34.2％， 48.9％， 53.9％， respectively. Compared with control group， mRNA expression of Collagen Ⅰ， TIMP-1 and protein expression of α-SMA were decreased significantly in protopine low-concentration， medium-concentration and high-concentration groups， while protein expression of MMP- 2 was increased significantly， with statistical significance （P＜0.05 or P＜0.01）. Apoptotic rate of HSC-LX2 cells and mRNA expression of MMP-2 were increased significantly in protopine medium-concentration and high-concentration groups， mRNA expression of α-SMA and Collagen Ⅲ， protein expression of Collagen Ⅰ and Collagen Ⅲ were decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. CONCLUSIONS： Protopine can induce the apoptosis of HSC-LX2 cells and inhibit their cell proliferation， and reduce the expression of a-SMA， Collagen Ⅰ， Collagen Ⅲ and TIMP-1， and increase the expression of MMP-2.

Study on Prevention Effect and Mechanism of Miao Medicine Liangjiang Weiyang Capsule on Gastric Ulcer Model Rats / 中国药房

OBJECTIVE： To study the protective effect and mechanism of Miao medicine Liangjiang weiyang capsule on gastric ulcer model rats. METHODS： Rats were randomly divided into normal group （normal saline）， model group （normal saline）， positive control group （omeprazole， 0.02 g/kg） and Liangjiang weiyang capsule low-dose， medium-dose and high-dose groups （0.3， 0.6， 1.2 g/kg）， with 12 rats in each group. All rats were intragastrically administered once a day for consecutive one week. 1 h after last administration， all rats except those in normal group were given the absolute ethanol to induce gastric ulcer model. 1 h after modeling， gastric juice volume， gastric juice pH， pepsin activity， gastric ulcer area and inhibitory rate of gastric ulcer were recorded in each group. Histopathological changes of gastric mucosa in rats of each group were observed by HE staining. The serum levels of TNF-α and IL-6 were determined by ELISA. The expression of nuclear factor-κB pathway related protein （p-NF-κB p65， p-IκBα） in gastric tissue of rats were determined by Western blot. RESULTS： Compared with normal group， gastric juice volume， pepsin activity， gastric ulcer area， TNF-α and IL-6 levels in serum， p-NF-κB p65 and p-IκBα levels in the gastric tissue were significantly increased/rised （P＜0.05）， while gastric juice pH was significantly decreased （P＜0.01）； there were gastric mucosal hyperemia and redness， obvious defect of mucosal epithelial cells， destruction of gland structure and incomplete cell structure. Compared with model group， gastric juice volume， pepsin activity， gastric ulcer area and the levels of TNF-α and IL-6 were reduced/decreased significantly in positive control group， Liangjiang weiyang capsule medium-dose and high-dose groups （P＜0.05 or P＜0.01）， while pH value of gastric juice was increased significantly （P＜0.05）； gastric mucosa was normal， gland destruction was alleviated and cell structure was intact. The levels of p-NF-κB p65 and p-IκBα in gastric tissue were significantly decreased in Liangjiang weiyang capsule high-dose groups （P＜0.05）. CONCLUSIONS： Liangjiang weiyang capsule play a role to protect gastric ulcer by increasing gastric juice pH， inhibiting pepsin activity， reducing the release of inflammatory factors as TNF-α， IL-6 and inhibiting the expression of NF-κB pathway related protein.

Safety and feasibility of enhanced recovery after surgery in perioperative management of pancreatectomy: a Meta analysis / 中华消化外科杂志

Objective To systematically evaluate the safety and feasibility of enhanced recovery after surgery (ERAS) in perioperative management of pancreatectomy.Methods Literatures were researched using CNKI,Wanfang database,VIP database,PubMed,Cochrane Library,Embase from January 1990 to March 2018 with the key words including "快速康复外科,加速康复外科,胰腺切除术,胰十二指肠切除术,惠普而术,ERAS,enhanced recovery,fast track,pancreatic surgery,pancreatectomy,Whipple,pancreatoduodenectomy,pancreatoduodenal resection".The cohort study about ERAS in elective pancreatic surgery or pancreaticoduodenectomy were received and enrolled.The patients using ERAS in perioperative management and using traditional perioperative management were respectively allocated into the ERAS group and control group.Two reviewers independently screened literatures,extracted data and assessed the risk of bias.Count data were described as odds ratio (OR) and 95％ confidence interval (CI).Weighted Mean Difference (WMD) was used as a consolidated statistics for measurement data that were measured using the same tool,and standardized mean difference (SMD) was used as a consolidated statistics for measurement data that were measured using the different tools.The heterogeneity of the studied was analyzed using the I2 test.Results Nineteen retrospective cohort studies were enrolled in the Meta analysis,and total sample size was 3 699 patients,including 1 823 in the ERAS group and 1 876 in the control group.The results of Meta analysis showed that there were statistically significant differences in the time of postoperative nasogastric tube removal,time for postoperative solid diet intake,time of postoperative defecation recovery,incidence of postoperative overall complications,incidence of postoperative delayed gastric emptying,incidence of postoperative intra-abdominal infection,duration of postoperative hospital stay and hospital expenses between ERAS group and control group (WMD=-1.70,-3.61,-0.86,OR =0.65,0.60,0.70,WMD=-4.64,SMD=-0.48,95％CI:-2.97--0.42,-4.70--2.53,-1.01--0.71,0.52-0.81,0.45-0.80,0.54-0.91,-5.91--3.38,-0.77--0.18,P ＜ 0.05).There was no statistically significant difference in the operation time,volume of intraoperative blood loss,incidence of postoperative pancreatic fistula,incidence of postoperative wound infection,readmission rate,reoperation rate and mortality between ERAS group and control group (WMD=-9.73,-14.39,OR=0.85,0.72,1.05,0.81,0.74,95％CI:-34.24-14.78,-116.96-88.17,0.72-1.01,0.46-1.14,0.83-1.32,0.58-1.13,0.53-1.02,P＞0.05).The results of subgroup analysis showed that heterogeneity of data was from eastern and western countries.Conclusion ERAS in the perioperative management of pancreatectomy is safe and feasible,it can also promote postoperative recovery of patients and reduce incidence of complications and financial burden.

Research advances in the role of mesenchymal stem cells and their exosomes in treatment of liver diseases / 中华肝脏病杂志

At present, liver transplantation remains the most effective treatment for acute liver failure and advanced cirrhosis, but its use and promotion are limited by insufficient organ donors, financial consideration, and requirements for lifelong immunosuppression. In recent years, stem cell transplantation has been recommended as an effective substitutive therapy for liver disease. Mesenchymal stem cells, also known as pluripotent interstitial stromal cells, are self-renewing cells that can be found in almost all postnatal organs and tissues, including the liver. Their potential to differentiate into hepatocytes and immunomodulatory properties provide new insights into the use of mesenchymal stem cells in the treatment of acute and chronic liver diseases. This article reviews the characteristics of mesenchymal stem cells, their mechanisms in the treatment of acute and chronic liver diseases, and related risks.

Study on the Preventive and Therapeutic Effect of Shuganning Injection on Alcoholic Liver Fibrosis in Rats / 中国药房

OBJECTIVE:To investigate the preventive and therapeutic effect of Shuganning injection on alcoholic liver fibrosis (ALF) in model rats,and provide experimental basis for its clinical application for alcoholic liver disease. METHODS:50 rats were enrolled and intraperitoneally given mixed liquid of 60% alcohol-corn oil-pyrazole to reduce ALF model. Another 10 rats were enrolled and intraperitoneally given normal saline,as normal control group. After 16 weeks,survived model rats(n=40)were ran-domly divided into model group,positive control group(Anluo huaxian pill 0.75 g/kg,ig),Shuganning injection high-dose,medi-um-dose,low-dose groups(4.8,2.4,1.2 mL/kg,ip),8 in each group. Normal control group and model group were intraperitone-ally injected equal volume of normal saline (5 mL/kg),administration groups were given relevant medicines,once a day,for 8 weeks;and modeling was contiuously conducted at the same time. After administration,body mass of rats was weighed,and the levels of liver function indexes [aspartate aminotransferase(AST),alanine aminotransferase(ALT)] and liver fibrosis indexes [hyal-uronic acid(HA),laminin(LN),type Ⅲ procollagen(PⅢNP),type Ⅳ collagen(Ⅳ-C)] in serum of rats were detected. Liver index of rats was determined and pathological changes of liver tissue were observed. RESULTS:Compared with normal control group,body mass of rats in model group was significantly decreased(P<0.05);liver index,and liver function index,liver fibro-sis index levels in serum were significantly increased (P<0.05 or P<0.01). Liver tissue showed steatosis,hepatocyte vacuoliza-tion,a large number of fibrous tissue deposition around portal areas and other pathological changes. Compared with model group,above-mentioned changes were improved significantly in administration groups (P<0.05 or P<0.01). CONCLUSIONS:Shugan-ning injection can obviously improve liver tissue damage of model rats with ALF,showing certain preventive and therapeutic effect on alcoholic liver disease.

Effect of Bunao Capsule on Learning,Memory and Antioxidative Abilities of Rats with Alzheimer′s Disease / 医药导报

Objective To investigate the effect of Bunao capsule on learning,memory and antioxidative abilities of rats with Alzhheimer’s disease(AD)induced by D-galactose combined with amyloid β-protein(Aβ25-35 ),and provide experimental basis for the prevention and treatemtn of AD. Methods A total of 90 SD male rats were randomly divided into model control group,piracetam group,sham operated group,Bunao capsule(0.79,1.58,3.15 g·kg-1 )groups(n= 15 each).The rat models were established by intraperitoneal injection of D-galactose and injection of Aβ25-35 into the bilateral lateral cerebral ventricle.Then rats were given corresponding drugs by gavage in different groups for 8 weeks.The learning and memory abilities were meseured by Morris water maze test.The morphology of brain cells was observed by HE staining.The activities of glutathione peroxidase(GSH-Px)and superoxide dismutase( SOD),and the malondialdehyde( MDA)contents in the brain tissues were measured by spectrophotometry. Results The target quadrant residence time was(20.39±7.75)s and(20.82±5.09)s in Bunao capsule (1.58,3.15 g·kg-1 )groups,which were significantly increased as compared with that in model control group[(12. 35 ± 6.95)s](P<0.01).Brain nerve cell morphology in Bunao capsule(1.58,3.15 g·kg-1 )groups was obviously improved as compared with that in model control group,and was close to that in sham operated group.The activities of GSH-Px and SOD were significantly increased,and MDA contents decreased in Bunao capsule groups as compared with those in model control group (P<0.01). Conclusion Bunao capsule can dose-dependently improve the learning,memory and antioxidative abilities of AD rats.The mechanism may involve upregulation of antioxidative enzyme activities and removal of oxidative products.

Levels of histone modifications in activated primarily cultured rat hepatic stellate cells / 中国病理生理杂志

AIM:To investigate the changes of histone modifications during the activation of primarily cultured rat hepatic stellate cells ( HSCs) and the relationship between histone modification patterns andα-smooth muscle actin (α-SMA) expression, and to explore the roles of histone modifications in the activation of HSCs.METHODS:The rat HSCs were isolated by in situ perfusion of collagenase combined with density gradient centrifugation, cultured in vitro and identi-fied by immunofluorescence staining.The morphological features of the cells were observed under inverted microscope.The changes of desmin and α-SMA during the activation of HSCs were detected by immunofluorescence staining and Western blotting.The levels of histone 3 lysine 4 dimethylation (H3K4me2), histone 3 lysine 9 dimethylation (H3K9me2), his-tone 3 lysine 9 acetylation (acH3K9) and histone 4 lysine 12 acetylation (acH4K12) in quiescent HSCs and activated HSCs were determined by Western blotting.RESULTS: The morphology of HSCs shifted from a quiescent phenotype to highly activated myofibroblast during the culture.Immunofluorescence staining and Western blotting showed that the expres-sion levels of α-SMA and desmin were increased over time and reached maximum at 15 d.According to the results of cell morphology and immunofluorescence staining, the cells cultured for 24 h and 15 d were quiescent and activated HSCs, re-spectively.Compared with quiescent HSCs, there were higher H3K4me2 and lower H3K9me2, acH3K9 and acH4K12 modification levels in activated HSCs ( P<0.01 ) .CONCLUSION: Histone modifications show anomalous expression during the activation of primarily cultured rat HSCs.Histone modifications may contribute to the transdifferentiation of HSCs and the development of hepatic fibrosis.

Comparison of acute toxicity of extract of unprocessed indian atringbush root and its two different processed products / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the acute toxicity and content of daphnoretin among extracts of unprocessed indian string-bush root and its two different processed products, and to provide a basis for discussion of the mechanism of two processed methods.</p><p><b>METHOD</b>Extracts of unprocessed indian stringbush root and processed indian stringbush root with "sweat" and "artificial sweat" were prepared. The mice were intragastrically administrated once with these three extracts, the mortalities of mice were observed, and the median lethal dose (LD50) of different extracts were calculated with Bliss method. The determination of daphnoretin in these three samples was performed by high performance liquid chromatography.</p><p><b>RESULT</b>The LD50 of indian stringbush root extracts, indian stringbush root processed with "sweat" and with "artificial sweat" were 46.678, 72.190, 67.953 g x kg(-1), respectively. The contents of daphnoretin in unprocessed indian stringbush root, indian stringbush root processed with "sweat" and with "artificial sweat" were 0.189%, 0.407% and 0.345%, respectively.</p><p><b>CONCLUSION</b>The toxicity of indian stringbush root processed with both "sweat" and "artificial sweat" is lower than that of the original rude drug. But the decreasion of toxicity of processed products is not by the reduced daphoretin content.</p>

Virulence analysis between environmental and clinical isolates of Cryptococcus neoformans var. grubü based on microsatellite patterns / 中华微生物学和免疫学杂志

Objective To investigate the genetic relation between Cryptococcus neoformans var.the clinical strains in MLMT - 13 genotype and the environmental strains in MLMT - 36 genotype. Methods Multilocus microsatellite typing (MLMT) method was applied for the genotype analysis in our study.Through this method, we recognized two genotypes that distinguish a majority of clinical and environmental strains. In order to compare virulence between the two types, we chose to infect BALB/c mice (6 weeks,female) with 9 MLMT-13 strains and 10 MLMT-36 strains intravenously. Results Forty( 17 clinical and 23 environmental isolates) were analyzed. Of 17 clinical strains, 9 belonged to a major type of MLMT-13 (52.9%). They were mainly isolated from clinical specimens. About 43.5% of strains from the environment belong to a major type of MLMT-36, which are indigenous to environments and which were not isolated from clinical samples. The mortality rate and pathological changes of the above mice were observed during two months after injection. The results showed that the mortality rate of mice infected with MLMT-13 strains was 100%, while the mortality rate with MLMT-36 strains was 7. 5%. The pathological sections showed that lesions of MLMT-13 infected mice appeared in the brain, lungs, liver and kidneys, while the lesions of MLMT-36 infected mice only appeared in the brain. Most brains of MLMT-13 infected mice were distorted,and both the number and size of lesions in such brains were much larger than those of MLMT-36 infected mice. Conclusion Our study illustrated the virulent difference between MLMT-13 and MLMT-36, which are isolated from patients and environment respectively. The results inferred that some genetic changes, such ss microsatellite repeats, might occur between environmental and clinical isolates through their environmental adaptation progress.

Isolation, culture, induction and differentiation of fetal bone marrow-derived postembryonic pluripotent stem cells / 中国组织工程研究

BACKGROUND: At present, studies show that a kind of stem cell community which in many kinds of organizations can differentiate into tissue cells of different embryonic layers; but those are different from embryonic stem cells, embryonic stem cell will lose the part differentiation potential gradually during the development of pregnancy, and will present some special phenotypes or the molecular markers, as CD105 and so on, will cell it postembryonic pluripotent stem cells.OBJECTIVE: To study the isolation of postembryonic pluripotent stem cells from fetal bone marrow, proliferative culture in vitro, induction and differentiation; transplantation to the liver of SCID mice with hepatic failure, and detect therapy effects.DESIGN, TIME AND SETTING: Cell observation and animal randomization experiment which was completed in the Ministry of Health of Cell Engineering Technology Research Center, Tianjin Third Central Hospital from March 2003 to March 2005.MATERIALS: The postembryonic pluripotent stem cells were extracted from thighbone and shinbone of 22-week old fetuses under sterile circumstance. Adult female SCID mice ware regarded as the recipients. CD105 immunornagnetic beads were provided by Miltenyi Biotec, Germany; mouse-anti-human albumin by Sigma, USA; basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF) by PEPROTECH, UK.METHODS: Postembryonic pluripotant stem cells obtained from fetal bone marrow were isolated using density gradient centrifugation and micromagnetic beads technique. The hepatocyte-like cells were induced and differentiated with culture media containing HGF (30 ng/mL) and bFGF (20 ng/mL). Twenty-four SCID mice were randomly divided into experimental group and control group with 12 mice in each group. Hepatic injury models were established with intraperitoneal injection of D-galactosamine On the next day, about 106 CD105(+) cells were perfused into liver in situ in the experimental group, and about 106 CD105(-) cells or isovolumic culture medium were perfused in the control group.MAIN OUTCOME MEASURES: Two, seven days, one and three months after the transplantation of cells, human albumin expression in the liver tissue was detected by immunohistochemistry.RESULTS: The immunocytochemical assay of the cells after micromagnetic beads selection showed that the CD105 expression was slightly positive; the doubling time of the cells in the logarithmic growth period was around 30 hours; after being expanded for 10 population doublings, the cells entered decline period. The cells were transplanted into SCID mice's liver; 3 months later, the human serum albumin in the mouse liver was assessed by using monoclonal antibody of mouse-anti-human serum albumin, dotted or small focal expression of the protein could be detected. However, any expression was not observed in the control group.CONCLUSION: The bone marrow-derived pluripotent stem cells are able to transform to hepatocytas in the hepatic microenvironment.

Development in the study on seed cell resources of liver tissue engineering / 国际生物医学工程杂志

Liver tissue engineering is one of the important subjects in tissue engineering field.The major goal of the research is to treat end-stage liver failure and liver based inborn errors of metabolism.Two strategies are usually employed:seed cell transplantation for liver tissue repair and regeneration of "liver tissue" consisted of an extraeorporeal bioreactor loaded with cenular component.The seed cells is one of the key ingredient of liver tissue engineering,which include adult hepatocytes,various kinds of liver stem cells,and immortalized cell fines,etc.However,to date,there is no optimal cell resource for application.Many problems should be solved before these cells can be widely used in clinic.This review focuses on the research regarding the seed cells and their potential for the application in liver tissue engineering.

Design of a hollow-fiber bioreactor and perfusion study in vitro / 生物医学工程学杂志

This article presents the design of a bioreactor using hollow fiber membrane and, isolated hepatocytes of suckling pigs, and the experimental study of its efficacy in vitro. Liver cells were harvested from suckling pigs with collagenase perfusion in situ, and parenchymal and non-parenchymal hepatocytes were cocultured in a hollow fiber module which was rotated sporadically. Bioartificial liver(BAL) was developed using this bioreactor,and the BAL was perfused with ascites of patients suffering from liver cirrhosis. The yield of viable hepatocytes was (6.29 +/- 0.37) x 10(8) cells, and cell viability was greater than 84%. Hepatocytes aggregated to multi-cells spheroids after being rotated every thirty minutes for three hours. The hepatocytes in the bioreactor could synthesize urea. Total billirubin was decreased, and AST was significantly increased in the group of bioreactor, as compared with that in the control group. Glucose decreased in the group of bioreactor,whereas there was no significant descent in the control group; and the difference between the two groups was significant. The above results demonstrate that this bioreactor is effective for decreasing total bilirubin and glucose.

Development of hepatocyte transplantation / 国际生物医学工程杂志

As a safe and effective treatment method of fulminic hepatic failure and end stage liver disease, hepatocyte transplantation has been widely verified in many animal experiment and clinical research. It has many advantages. For instances, It is safe and easy to perform. It causes little trauma and is convenient for retransplantation. The hepatocytes are less immunogenetic and the cells from one donor liver can be used for several patients, and so on. The cell source, route of transplantation, theoretical basis and clinical application of the method are reviewed in this paper.