For more than 40 years primary hepatocytes culture technique has been utilized extensively for assessing effects of drug on metabolizing enzymes (especially cytochromes P450), drug metabolism, drug-drug interactions, and the mechanisms of cytotoxicity and genotoxicity. Human derived primary hepatocytes reserve the metabolism function and enzyme activity of liver. Therefore, this technique has been widely used as a reliable and efficient tool in the drug and xenobiotics evaluation and screen in vitro. This review focuses on primary culture technique of hepatocytes and its application in drug metabolism and toxicology research and evaluation.

Objective:The significant role of long non-coding RNAs (lncRNAs) in early diagnosis and predicting prognosis has been recognized in various cancers recently.However,the prognostic value of HOXA transcript at the distal tip (HOTTIP),a vital lncRNA in tumorigenesis,remains unclear.In this study,we evaluated its prognostic value by analyzing the correlation of HOTTIP expression with overall survival (OS),lymph node metastasis (LNM) and distant metastasis (DM) in different cancer types by meta-analysis.Methods:We performed a systematic search in PUBMED,MEDLINE,Web of Science and Cochrane Library update to November of 2016.A total of 604 patients from 6 studies were included in final analysis and went through a quantitative meta-analysis by Review manager 5.3.Results:We demonstrated that high expression of HOTTIP had a significant correlation with poor OS (hazard ratio [HR] =2.37,95％ confidence interval [CI] =1.81-3.10,p＜0.001),high LNM rate (odds ratio [OR]=2.29,95％CI=1.54-3.40,p＜0.001) as well as more DM occurrence (OR=3.30,95％CI=1.78-6.12,p＜0.001).Conclusion:Our results indicated that long non-coding RNA HOTTIP may serve as a potential prognostic biomarker in cancer progression.

Objective To study the anti-neuroglioma effect of methyl-mercuric chloride(MMC) by observing the morphological changes of apoptotic neuroglioma cells induced with MMC in rats with brain neuroglioma.Methods The rat models of neuroglioma were established,and divided into two groups.The rats in experimental group were lavaged with MMC 1 week after injected with C6 glioma cells,10 mg?kg-1every day,the rats in control group were treated with sodium chloride at the same dose.24 d after inoculation all rats were sacrificed except natural death,the brain tissues were obtained,and the pathohistological changes were observed under light microscope and transmission electron microscope.Results The macropathological result showed that the tumor volume in experimental group was smaller than that in control group.Under light microscope,in experimental group the growth density of C6 ghioma cells was lower than that in control group,and the apoptotic cells with smaller volume and karyopyknosis were found.The result of transmission electron microscope showed that in experimental group,the glioma cells had some changes such as karyopyknosis,chromoplasm margination,nuclear fragmentation and vacuolar degeneration and so on.Conclusion MMC has inhibitory effect on the proliferation of C6 glioma in rats in vivo,its mechanism may be related to inducing the apoptosis of neuroglioma cells.

Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Fibrinolysis ; drug effects ; Glycoproteins ; blood ; Humans ; Injections ; Male ; Middle Aged ; Phytotherapy ; Plasminogen Activator Inhibitor 1 ; blood ; Platelet Aggregation ; drug effects ; Pulmonary Heart Disease ; blood ; drug therapy ; Tissue Plasminogen Activator ; blood

Adult ; Aged ; Aged, 80 and over ; Conjunctival Neoplasms ; diagnostic imaging ; pathology ; radiotherapy ; surgery ; Female ; Follow-Up Studies ; Humans ; Lymphoma, B-Cell, Marginal Zone ; diagnostic imaging ; pathology ; radiotherapy ; surgery ; Male ; Middle Aged ; Orbital Neoplasms ; diagnostic imaging ; pathology ; radiotherapy ; surgery ; Prognosis ; Retrospective Studies ; Tomography, X-Ray Computed

OBJECTIVETo explore the clinical effects of transforaminal endoscopic spine system in surgical revision of lumbar vertebrae.

METHODSFrom January 2012 to October 2013,14 patients who needed reoperations of lumbar vertebrae were treated using transforaminal endoscopic spine system (TESSYS). There were 8 males and 6 males, aged from 27 to 84 years old with an average of (50.4 ± 18.9) years. Visual analogue scale (VAS) and Japanese Orthopaedic Association Scores (JOA) were compared before and after surgical revision. Macnab standard was used to assess the clinical effect.

RESULTSAll the patients were followed up from 6 to 27 months with the mean of 18 months. Preoperative VAS score was 6.79 ± 1.31, and in a week,3 months and 6 months after operation were 2.50 ± 1.29, 2.21 ± 1.53, 1.64 ± 1.08, respectively, which were all much lower (P < 0.01) than preoperative score. Preoperative JOA score was 12.43 ± 1.95, and the above corresponding postoperative JOA scores were 21.50 ± 3.78, 21.93 ± 4.55, 23.36 ± 4.33, respectively, which were all much higher than preoperative score (P < 0.01). According to the modified Macnab criteria, 5 patients got an excellent results, 7 good, 1 fair and 1 poor. The nerve root injury of L5 occurred in 1 case during paracentesis and no other complications were found.

CONCLUSIONSelecting the appropriate indications using TESSYS in surgical revision of lumbar vertebrae can successfully avoid the operation scar, reduce the surgical complications and obtain satisfactory clinical outcomes.

Adult ; Aged ; Aged, 80 and over ; Endoscopy ; methods ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Reoperation ; Spinal Fusion ; methods

two-dimensional strain echocardiography can observe dynamic change of regional systolic function,combined with the two methods can more accurately assess the treatment results of coronary artery bypass surgery.

Objective To investigate the effect of mild hypothermia on glucose regulated protein 78 (GRP78) expression in hippocampus following transient cerebral ischemia-reperfusion (I/R) in aged rats.Methods One hundred and forty-four male aged Sprague-Dawley rats,weighing 450-550 g,were randomly divided into 3 groups (n=48 each): sham operation group (group S),I/R group and mild hypothermia group (group H).Global cerebral I/R was induced by 4-vessel occlusion method.Bilateral vertebral arteries were permanently occluded by cauterization,and bilateral carotid arteries were occluded for 5 min.The surface cooling was started immediately after reperfusion and maintained for 3 h.During surface cooling,the body temperature was maintained at 32-34 ℃(rectal).At 6,12,24 and 48 h of reperfusion,the expression of GRP78 was determined using immuno-histochemisty and Western blot,and the viable neurons in CA1 area were counted.Results The number of viable neurons was significantly smaller at each time point,and the expression of GRP78 was significantly higher at 6,12 and 24 h of reperfusion,while lower at 48 h of reperfusion in I/R group,and the number of viable neurons was significantly smaller,and the expression of GRP78 was significantly higher at each time point in group H than in group S (P＜ 0.05).The number of viable neurons was significantly larger at 12,24 and 48 h of reperfusion,and the expression of GRP78 was significantly higher at each time point in group H than in group I/R (P ＜ 0.05).Conclusion Mild hypothermia can further up-regulate GRP78 expression in hippocampus following transient cerebral I/R in aged rats,thus reducing endoplasmic reticulum stress and cerebral I/R injury.

Objective To dynamically observe the changes of cytokines of splenocytes in mice immunized with recombinant bifidobacteria bifidum (Bb)- Eg95-EgA31 vaccine of Echinococcus grauulosus (Eg). Methods Balb/c mice were vaccinated by 5× 108 colony forming unit(CFU) orally and 5 × 105 CFU intranasally, respectively.Mice were killed on week 0,2,4,6,8,10, 12,14,16, 18 and 20 after immunization, respectively, and spleens were separated for cell culture with the stimulation of EgAg, concanavalin A (ConA) or lipopolysaccharide (LPS). The splenocyte supernatants were collected to determine the levels of interferonγ(IFN-γ), interleukin(IL)-12, tumor necrosis factor α(TNF-o) and IL-l0 using enzyme linked immunosorbent assay(ELISA) with MRS as control. Results In the oral immunization group, the levels of IFN-γ, IL-12, TNF-α and IL-10 showed a significant increase from week 2 to week 8, week 2 to week 8, week 4 and week 6 to week 10 after vaccination, respectively, and reached the highest level on week 4, week 2, week 4 and week 6 after vaccination, respectively;in EgAg stimulation group, the levels of IFN-γ, IL-12, TNF-α and IL-10 were (700.0 ± 115.5), (45.0 ± 5.8), (350.0 ± 57.7), (112.5 ± 14.4)ng/L, respectively, compared with week 0[(35.0 ± 5.8), (12.5 ± 2.9), (190.0 ± 11.6), (25.0 ± 5.8)ng/L, P ＜0.05 or ＜ 0.01] and MRS control group[(37.5 ± 5.0),(13.8 ± 2.5), (195.0 ± 5.8), (27.5 ± 2.9)ng/L, P＜ 0.05or ＜ 0.01]. In the intranasal immunization group, the levels of IFN-γ, IL-12, TNF-α and IL-10 showed an obvious increase from week 2 to week 8, week 2 to week 8, week 2 to week 6 and week 6 to week 16 after vaccination,respectively, and reached the highest level on week 2, week 2, week 4 and week 8 after vaccination, respectively;in EgAg stimulation group, the levels of IFN-γ, IL-12, TNF-α and IL-10 were (700.0 ± 115.5), (55.0 ± 5.8),(275.0 ± 28.9), (140.0 ± 11.6)ng/L, compared with week 0[(35.0 ± 5.8), (12.5 ± 2.9), (190.0 ± 11.6), (25.0 ±5.8)ng/L, P ＜ 0.05 or ＜ 0.01] and MRS control group[(37.5 ± 5.0), (13.8 ± 2.5), (195.0 ± 5.8), (27.5 ± 2.9)ng/L, P ＜ 0.05 or ＜ 0.01]. The cytokine levels in the groups with EgAg, ConA or LPS stimulus were significantly higher than those in the corresponding splenocytes suspension groups(P ＜ 0.05 or ＜ 0.01) , and the cytokine levels in the groups with ConA or LPS stimulus were obviously higher than those in the corresponding groups with EgAg stimulation(P ＜ 0.05 or ＜ 0.01). Conclusion The mixed Th1 and Th2 type response can be induced in mice immunized with the recombinant Bb-Eg95-EgA31 vaccine of Echinococcus granulosus in the early stage of immunization(2 to 6weeks).

Objective To construct a recombinant plasmid pGEX-TSO45W-4B-TSOL18 of Taenia solium,to obtain purified recombinant protein TSO45W-4B-TSOL18 and to prepare rabbit antiserum against the recombinant fusion protein.Methods TSO45W-4B and TSOL18 encoding genes were connected with hydrophobic (Gly4Ser)3 linker.TSO45W-4B-TSOL18 fusion gene was synthesized and cloned into an expression vector pGEX-1λT to construct a recombinant plasmid pGEX-TSO45W-4B-TSOL18.The positive recombinants were transformed into Escherichia(E.) coli ArcticExpress (DE3),and the expression of recombinant protein was induced with isopropylβ-D-thiogalactopyranoside (IPTG) and analyzed by sodium dodecyl sulfate polyacrylamide gelelectrophoresis (SDS-PAGE).The expressed recombinant fusion protein was purified through affinity chromatography.The purified recombinant protein was mixed with complete Freunds adjuvant at a dosage of 0.5 mg.Rabbit was intramuscularly and subcutaneously immunized in the hind leg and the back,respectively.After 2 weeks the rabbit was boosted with purified recombinant protein which was mixed with incomplete Freunds adjuvant at the same dosage.Rabbit can be boosted every 10 days until an adequate response was achieved.At the 6th day after the last immunization,blood was collected from the rabbit heart.Serum was separated to purify and prepare the antiserum.ELISA was applied to determine the titer of the antiserum and Western blot assay was used to determine the specificity of the antiserum.Results The size of the synthesized fusion gene TSO45W-4B-TSOL18 was 789 bp.The results of restriction enzyme digestion showed that the recombinant plasmid pGEX-TSO45W-4B-TSOL18 was successfully constructed.DNA sequencing showed that the size of the fusion gene TSO45W-4B-TSOL18 was 789 bp,which was consistent with expected result.As demonstrated by SDS-PAGE,relative molecular mass of the expressed recombinant fusion protein was approximately 55 × 103,and its purity was 85％ after purification with affinity chromatography.Titer of the antiserum against the recombinant protein was 1 ∶ 512 000 in ELISA assay and the specific rabbit antiserum against the purified recombinant protein TSO45W-4B-TSOL18 could specifically bind to the recombinant fusion protein in Western blot assay.The relative molecular mass of the specific band was about 55 × 103,which was consistent with expected size.Conclusions The recombinant plasmid pGEX-TSO45W-4BTSOL18 of Taenia solium is successfully constructed.High quality recombinant fusion protein TSO45W-4B-TSOL18 and high titer rabbit antiserum are successfully prepared.