There are many kinds of diseases in hepatic surgery department including hepatitis,hepatic cirrhosis, liver cancer etl. Most of them are very ditficult to diagnose and treat, so it's necessary to change our traditional clinical practice teaching mode.Role-playing mode increases the interaction between students and teachers, which can achieve the desired effect with entertainment. So role-playing is a new model of clinical teaching as a meaningful innovation.

Chronic inflammation has been demonstrated closely related to the tumor progression.Tumorassociated macrophage,as the most abundant immune cells in the tumor microenvironment,is a key element that links inflammation and cancer.Recently,studies found that the phenomenon and function of tumor-associated macrophage almost tend to M2 type macrophage.As an important indicator,tumor-associated macrophage usually predicts the poor progress with the cancer development.In China,Most of patients with hepatocellular carcinoma are associated with chronic viral infection.A large number of macrophages in the liver infiltrated in chronic inflammation,which are differentiated by variety of mechanisms under the chronic inflammatory stimulation,promote the development of liver cancer.In this paper,we will review the tumor-associated macrophages and the development of liver cancer.

Objective:Human adipose-derived stem cells (hASCs)are a highly attractive source in bone tissue engineering.To generate a luciferase reporter system that could be used to quantitatively and rapidly examine osteogenic differentiation potential of human adipose-derived stem cells (hASCs ) in vitro,and eventually make it possible to monitor the osteogenic differentiation of transplanted cells in vi-vo.Methods:The genomic DNA harboring promotor regions of osteocalcin and DNA sequences encoding luciferase genes were amplified by PCR and cloned into the pLVX-pTRE-puro vector to generate the OCpro-Luc-Puro construct.Then,the OCpro-Luc-Puro construct together with three assistant vectors:pM-DLg/pRRE,pRSV-REV,and pVSVG,were transiently transfected into HEK293T cells followed by viral supernatants collection,filtration and concentration.Next,the hASCs stably expressing luciferase repor-ter gene driven by osteocalcin promotor were created with the lentivirus carrying OCpro-Luc-Puro cassette under puromycin selection.The OCpro-Luc-hASCs were then cultured in the absence or presence of osteo-genic differentiation medium.On the 7th and 1 4th days,after osteogenic induction,cellular extracts were collected and analyzed by luciferase reporter assay.Meanwhile,alizarin red staining and quantification as well as quantitative reverse transcription PCR (qRT-PCR)analysis of osteogenic associated genes osteo-calcin (OC),runt-related transcription factor 2 (Runx2)and alkaline phosphatase (ALP)were used to assess the osteogenic differentiation ability of OCpro-Luc-hASCs.Results:OCpro-Luc-Puro plasmid and OCpro-Luc-hASCs were successfully generated.On the 7th and 1 4th days after osteogenic induction,the luciferase activity of the cellular extracts from OCpro-Luc-hASCs was dramatically increased.Consistently, the extracellular matrix mineralization,as shown by Alizarin red S (ARS)staining and quantification was also markedly intensified and a marked increase of the mRNA expression levels of OC,Runx2 and ALP, although to variable extent,was observed upon osteogenic differentiation.These results indicated that the observations from traditional experiments examining hASCs osteogenic differentiation were largely in agreement with that of our luciferase reporter assay in OCpro-Luc-hASCs.Conclusion:We established a luciferase reporter system that could be used to rapidly,quantitatively and specifically determine osteo-genic differentiation ability of hASCs.Comparing with the traditional time-consuming methods,the system we generated here was highly effective.This system not only can be used to examine ostogenic differentia-tion of hASCs in a high throughput manner,but also provides a way to monitor ostogenic differentiation of cells in vivo.

Objective:To construct and evaluate a novel tissue-engineered bone composed of murine stromal cell-derived factor 1(mSDF-1), simvastatin (SIM) and collagen scaffold (Bio-Oss?), serving as a cell-homing approach for bone formation .Methods: In the study , 32 ICR mice were randomly divided into 4 groups,each group including 8 mice.The drug-loaded collagen scaffolds were implanted subcutaneously onto the cranium of each mouse according to the groups: ( 1 ) 1 ∶50 ( volume ratio ) dimethyl sulfoxide ( DMSO ) /phosphate-buffered saline ( PBS ) solution +collagen scaffold ( blank control group ); ( 2 ) 10 -3 mol/L SIM solution +collagen scaffold ( SIM group ); ( 3 ) 200 mg/L mSDF-1solution +collagen scaffold (mSDF-1 group); and (4) 10 -3mol/L SIM +200 mg/L mSDF-1 solution +collagen scaffold ( SIM +mSDF-1 group) .One week after implantation , the mice were trea-ted by injecting the same drug solution mentioned above around the scaffold once a day for two days .The specimens were harvested 6 weeks after implantation and the bone formation was evaluated by soft X-ray analysis , HE staining and immunohistochemical staining .Angiogenesis of each group was checked by calculation of vessels in each tissue section .Results:Six weeks after implantation , the collagen scaffolds were retrieved.The value of gray scale for the SIM +mSDF-1 group[(421 836.5 ±65 425.7) pixels] was significantly higher than that of the blank control group [(153 345.6 ±45 222.2)pixels, P<0.01], the SIM group [(158 119.2 ±100 284.2) pixels, P<0.01], and the mSDF-1 group[(255 529.5 ± 152 142.4) pixels, P <0.05 ]; HE staining analysis revealed that significant bone formation was achieved in the SIM +mSDF-1 group; The immunohistochemical staining showed the existence of os-teopontin and osteocalcin in the SIM +mSDF-1 group; There were more vessels in the SIM +mSDF-1 group[(46 ±8)vessels/mm2] than in the blank control group [(23 ±7) vessels/mm2, P<0.01], and the SIM group[(24 ±6) vessels/mm2 , P<0.01].Conclusion:The novel tissue-engineered bone com-posed of mSDF-1, SIM and collagen scaffolds has the potential to form bone subcutaneously in vivo.It re-presents a novel method of in vivo bone re-generation without seed cell delivery .

Xiaochaihu Decoction comes from Shang Han Lun, which is a main formula for Shaoyang diseases. According to the principle of prescriptions corresponding to syndromes, the clinical application of modified Xiaochaihu Decoction for the treatment of swelling, pain, nausea, vomiting, bloating, and constipation after artificial total knee arthroplasty can achieve good efficacy.

Objective:To construct a novel biomimetic calcium phosphate (BioCaP) scaffold loaded with bone morphogenetic protein-2 (BMP-2),and to investigate its role in the osteogenesis of human adipose-derived stem cells (hASCs) in vitro and in vivo.Methods:The BioCaP scaffold coprecipitated with BMP-2 (BMP-2-BioCaP) was constructed in this study.Field emission scanning electron microscopy (SEM) was used to analyze the morphology of the surfaces.The release kinetics was measured to evaluate the slow-release characteristics in vitro.BMP-2-BioCaP was immersed in proliferation medium (PM) or osteogenic medium (OM),respectively.The supernatants were collected and used to culture hASCs in vitro.Cell numbers were determined using the cell-counting kit-8 (CCK-8) to assess the cell proliferation.After 7 and 14 days,alkaline phosphatase (ALP) staining and quantification were performed to test the activity of ALP.After 14 and 21 days,the calcification deposition was determined by alizarin red S (ARS) staining and quantification.The expressions of the osteoblast-related genes were tested on day 4 and day 14.In the in vivo study,6 nude mice were used and implanted subcutaneously into the back of the nude mice for 4 groups:(1) BioCaP scaffold only,(2) BioCaP scaffold + hASCs,(3) BMP-2-BioCaP scaffold,(4) BMP-2-BioCaP scaffold + hASCs (test group).After 4 weeks of implantation,hematoxylin-eosin (HE) staining was performed to evaluate the in vivo osteogenesis of hASCs.Results:SEM observations showed that BioCaP and BMP-2-BioCaP scaffold were entirely composed of straight,plate-like and sharp-edged crystal units,and the length of the crystal units varied between 5 and 10 μm.Release kinetics analysis demonstrated that BMP-2 incorporated with BioCaP could be released at certain concentration and last for more than 21 days,and the accumulative protein release could reach 20％.CCK-8 assays showed that cell proliferation was not significantly affected by BMP-2BioCaP.ALP activity was higher by the induction of OM + BMP-2-BioCaP than of the other groups (P ＜0.01).More mineralization deposition and more expressions of osteoblast-related genes such as Runt-related transcription factor 2 (RUNX2),ALP,osteopontin (OPN) and osteocalcin (OC) were determined in the OM + BMP-2-BioCaP group at different time points (P ＜0.01).HE staining showed that,in the test group and BMP-2-BioCaP scaffold group,the extracellular matrix (ECM) with eosinophilic staining were observed around hASCs,and newly-formed bone-like tissues could be found in ECM around the scaffold materials.Moreover,compared with the BMP-2-BioCaP scaffold group,more bone-like tissues could be observed in ECM with typical structure of bone tissue in the test groups.No obvious positive results were found in the other groups.Conclusion:BMP-2-BioCaP scaffold could achieve slow-release of BMP-2 and promote the osteogenic differentiation of hASCs in vitro and in vivo.The novel tissue-engineered bone composed of hASCs and BMP-2-BioCaPis promising for the repair of bone defect.

Objective To investigate the expressions of extracellular matrix protein 1 (ECM1)and matrix metalloproteinase-9 (MMP-9) in hepatocellular carcinoma (HCC) and to explore their correlations with metastasis and recurrence.Methods Immunohistochemisty was employed to determine the expressions of ECM1 and MMP-9 in 120 HCC and 17 normal liver tissues.Results The positive rates of ECM1 and MMP-9 expression in HCC tissues were 73.3％ (88/120) and 65.0％ (78/120),respectively.They were significantly higher than that in normal liver tissues.The expression of ECM1 in HCC tissues was significantly correlated with vascular invasion and TNM stage.The expression of MMP-9 in HCC tissues was significantly correlated with tumor size,number of tumor nodules,differentiation of tumor and degree of cirrhosis.They were not correlated with age,gender,serum AFP level,HBsAg,Child-Pugh class and capsulated tumor.Patients with ECM1 and MMP-9 positivity had significantly poorer overall and disease-free survival rates.There was a significant positive correlation between ECM1 and MMP-9 (r=0.585,P＜0.001).Conclusion Both ECM1 and MMP-9 were significantly related to invasion and metastasis.Over-expression of both ECM1 and MMP-9 had predictive values for prognosis and recurrence of HCC after surgery.

Objective To observe vasculogenic mimicry formation difference in human hepatocellular carcinoma cell lines MHCC97-H/L with different metastatic potentials under three-dimensional culture condition,and to study extracellular matrix and adhesion molecules-related mechanism of vasculogenic mimicry.Methods Three-dimensional cell culture system of MHCC97-H/L was established to observe tubelike structures by inverted microscope.Human umbilical vein endothelial cell(HUVEC),human hepatocellular carcinoma cell line Hep3B with non-metastatic potentials and normal human hepatic cell line HL-7702 were taken as control.Gene expression profiles of MHCC97-H and MHCC97-L were detected by Oligo extracellular matrix and adhesion molecules microarray analysis.Two differentially expressed genes were verified with semi-quantitative reverse transcriptionpolymerase chain reaction(RT-PCR)and Western blot.All data were analyzed using two sample t test.Results After a 24-hour three-dimensional culture,the length of tubelike structures was(474 ± 16)mm/cm2 in MHCC97-H,which were significantly longer than(320 ±41)mm/cm2 in MHCC97-L(t =6.119,P ＜0.05).Hep3B and HL-7702 could not form tubelike structures.Compared with MHCC97-L,the expression of 7 genes were up-regulated and the expression of 3 genes were down-regulated among a total of 113 angiogenesis genes in MHCC97-H.Two of the genes with differential expressions including tenascin C and extracellular matrix protein 1 were further validated by RT-PCR and Western blot.Conclusion MHCC97-H has stronger ability to form vasculogenic mimicry than MHCC97-L,and this perhaps correlates with the differential expression of certain extracellular matrix and adhesion molecules-related genes in MHCC97-H.

Hepatectomy for huge hepatocellular carcinoma (HCC) is difficult due to its huge size and the compression and invasion to the surrounding tissues as well as the important vascular systems.Surgical resection of huge carcinoma in the caudate lobe is a big challenge for hepatobiliary surgeons because of its special location and complex anatomical structure.As the improvement of surgical techniques in recent years,especially the promotion of the concept of precision liver surgery,many surgeons begin to take the challenge of resection of huge HCC in the caudate lobe in a variety of ways.In April 2014,a male patient aged 58 years with huge HCC in the caudate lobe was admitted to the Anhui Provincial Hospital.Precision right hemihepatectomy combined with caudate lobectomy was performed on this patient without occlusion of the hepatic inflow,and the efficacy was satisfactory.The key techniques involved in this procedure were discussed in this article.

Ischemia reperfusion injury is an important factor which has been affected the recuperation of hepatic function after hepatectomy and liver transplantation,and is a complex course in pathophysiology with many factors.With the development of research on ischemia reperfusion injury,effective prevention measures of ischemia reperfusion injury also have made new progress.And this will greatly improve the prognosis of hepatic surgery.The mechanism and its prevention measure of hepatic ischemia reperfusion injury were reviewed in this paper.