AIM,To verify whether p73;a homologue of p53,which supposed]y acts as a tumor suppressor gene in neuroblastoma,might also be a tumor suppressor in non-small cell lung cancer.METHODS:The allelic expres-sion of p73 in the six non-small cell lung cancer cell lines was studied by Sty I polymorphism analysis.The P73 gene ex.pressions in these six cell lines were examined by reverse transcription-PCR,the expressions of P73 protein in the five cell lines inducing tumor8 were also determined by immunohistochemistry.RESULTS:Homozygous allelic expression was dem.onstrated in all six cell lines and the GC/GC genotype Was the predominant type.Complete loss of the p73 expression both at mRNA and the protein level was revealed.CONCLUSION:Taken together,our data suggest that p73 might play a role as a tumor suppressor gene in human non-small cell lung cancer cell lines.

AIM:Toinvestigatetheassociationbetweenmethylationstatusofapoptosis-relatedgenesandche-mosensitivity in the lung adenocarcinoma cell line P 15.METHODS: Methylation-specific PCR was applied to detect the methylation status of p73, p14ARF, p16INK4a and bax genes of P15 cells in untreated control group and decitabine (DAC) treatment group.RT-PCR was used to detect the expression of p 73, bcl-xL, bad, bax, p14ARF and p16INK4a at mRNA level. Colony formation assay and cell growth inhibition assay were used to detect the sensitivity of P 15 cells to cis-diaminedichlo-roplatinum ( C-DDP) before and after DAC treatment .DAPI staining was used to determine the apoptosis of P 15 cells ex-posed to C-DDP before and after DAC treatment .RESULTS:p73, p16INK4a and bax were expressed in the methylation sta-tus.After DAC treatment, p16INK4a expression was decreased , and the expression of p73 and bax disappeared .The expres-sion of p73, p16INK4a and bax in the unmethylated status was weak , but the enhanced expression was observed following DAC treatment.After P15 cells were treated with DAC and C-DDP, the colony formation rate of the P15 cells was signifi-cantly decreased as compared with untreated control group .The apoptotic P15 cells in DAC+C-DDP treatment group were significantly higher than those in untreated control group (P<0.05).CONCLUSION:After treated with DAC, the sensi-tivity of P15 cells to C-DDP is increased due to the activation of silenced pro-apoptotic genes .DAC and C-DDP synergisti-cally promote tumor cell apoptosis .They have significant anti-tumor effect .

Cell therapy has a very promising potential for end-stage liver diseases (ESLD).Fetal liver stem/progenitor cells (FLSPCs) have advantages of safety,high survival and proliferation rates,and a small volume,all which make them ideal for liver disease stem cell therapy.During the early phase of our study,we applied a three-step separation method to enrich FLSPCs and obtained a separation efficiency similar to that of the flow-cell sorting method.Additionally,using a fulminant hepatic failure model in rats,we have demonstrated that FLSPCs can contribute to morphological and functional recovery of the liver.This manuscript will discuss how FLSPCs can be induced to accurately differentiate into hepatocytes and cholangiocytes and how FLSPCs maintain self-renewal.The Notch signaling plays a critical role in regulating the differentiation and self-renewal of many types of stem cells.Our previous findings have shown that the Notch signaling plays an important role in FLSPCs differentiation into hepatocytes.Therefore,the Notch signaling might be involved in the differentiation and self-renewal of FLSPCs.We conducted a study on the regulatory effects and relative molecular mechanisms of the Notch signaling on FLSPCs and found the corresponding interfering target,which might become an index for the clinical application of FLSPCs.

Objective Proliferation and apoptosis play a major role in the development of tumor cells,and the intranuclear transcriptional factor c-fos is significantly up-regulated in the primary hepatocellular carcinoma and involved in early carcinogenesis.The purpose of the present study is to investigate the apoptotic effect of c-fos antisense oligodeoxynucleotide(ASO) on human hepatocellular carcinoma HepG2 cells and the participation of Caspase3 in this process.Methods Cell culture,Hoechst 33258 staining,real-time PCR and Western blotting were used in present study.Cultured HepG2 cells were divided into 3 groups: 1) control group: cultured with 10?l saline;2) sense oligodeoxynucleotide(SO,used as a negative control) treated group: co-cultured with 10?l SO(5?g/?l);3) ASO treated group: co-cultured with ASO 10?l(5?g/?l).The subsequent experiments were performed 1h after cultivation for each group.Hoechst 33258 staining was performed to detect the apoptosis by observing the staining of nuclear chromatin.Real-time PCR and Western blotting were used respectively to detect the expression of Caspase 3 at mRNA and protein levels after different treatments.Results Hoechst 33258 staining revealed that the nuclei of HepG2 cells showed diffuse and adqulis fluorescence in control and SO-treated groups,while dense and dark fluorescence was observed in ASO-treated group,which indicated that c-fos ASO had significantly induced apoptosis of HepG2 cells.The expression of Caspase 3 in ASO group was enhanced both at mRNA and protein levels compared to that in control groups.Conclusions C-fos ASO significantly induces apoptosis in human hepatocellular carcinoma HepG2 cells,as shown by Hoechst 33258 staining and higher expression of Caspase 3 mRNA and protein.Moreover,Caspase 3 activation is involved and probably plays an important role in c-fos ASO-induced apoptosis of HepG2 cells.

Objective To investigate the effect of microvascular invasion (MVI) on tumor recurrence of hepatocellular carcinoma (HCC) patients after hepatectomy,the efficacy of sequential transcatheter arterial chemoembolization (TACE) on positive MVI patients after hepatectomy,and the effect of comprehensive treatment on the prognosis of patients with tumor recurrence.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 136 HCC patients who underwent hepatectomy in the First Affiliated Hospital of Air Force Medical University from February 2015 to December 2016 were collected.Patients were treated with TACE,radiofrequency ablation (RFA) and molecular-targeted drugs after hepatectomy,and patients with tumor recurrence selected 1 or 2 above treatments.The patient received postoperatively outpatient reexaminations every 3 months to detect tumor recurrence and survival.Follow-up was from the day of the surgery to death or December 2017.Observation indicators:(1) treatment after hepatectomy;(2) effect of MVI on tumor recurrence of HCC patients after hepatectomy:clinicopathological features and tumor recurrence rate between positive and negative MVI patients;(3) efficacy of TACE on positive MVI patients:clinicopathological features and tumor recurrence rate in positive MVI patients with or without TACE;(4) effect of comprehensive treatment on the prognosis of patients with tumor recurrence.Measurement data with normal distribution were represented as x±s and analyzed using the independent-samples t test.Comparisons of count data were analyzed using chi-square test or Fisher exact probalility.The tumor recurrence rate and survival rate were calculated by the Kaplan-Meier method,and Log-rank test was used for survival analysis.Results (1) Treatment after hepateetony:of 136 patients undergoing hepatectomy,117 underwent single hepatectomy and 19 combined sequential TACE;59 had HCC recurrence,including 22 receiving comprehensive treatment.(2) Effect of MVI on tumor recurrence of HCC patients after hepatectomy:① Clinicopathological features:of 117 patients undergoing single hepatectomy,positive MVI was detected in 49 patients,including 44 males and 5 females,with an age of (52-± 10)years old;49 patients were in Child-Pugh grade A,including 36 combined with liver cirrhosis and 13 without liver cirrhosis;positive and negative alpha-fetoproteins (AFPs) were respectively detected in 34 and 15 patients.Negative MVI was detected in 68 patients,including 54 males and 14 females,with an age of (55± 11)years old;65 and 3 patients were respectively in Child-Pugh grade A and B,including 52 combined with liver cirrhosis and 16 without liver cirrhosis;positive and negative AFPs were respectively detected in 39 and 29 patients.There was no statistically significant difference in gender,age,Child-Pugh score of liver function,liver cirrhosis and comparison of AFP between positive and negative MVI patients (x2 =2.258,t =-1.626,x2 ＝ 0.804,0.138,1.758,P＞0.05).② Tumor recurrence rate:The 6-month and 1-year tumor recurrence rates after hepatectomy were respectively 30.77％,30.61％ in 117 patients undergoing single hepatectomy and 42.86％,51.02％ in 49 of 117 patients with positive MVI and 22.06％,27.94％ in 68 of 117 patients with negative MVI,showing statistically significant differences in 6-month and 1-year tumor recurrence rates between positive and negative MVI patients (x2 =5.738,6.465,P＜0.05).(3) Efficacy of TACE on positive MVI patients:① Clinicopathological features of 56 patients with positive MVI,7 received postoperatively sequential TACE,including 7 males,with an age of (50±4) years old;56 patients were in Child-Pugh grade A,including 5 combined with liver cirrhosis and 2 without liver cirrhosis;positive and negative AFPs were respectively detected in 2 and 5 patients.Forty-nine patients didn't combine sequential TACE,including 44 males and 5 females,with an age of (52± 10)years old;49 patients were in Child-Pugh grade A,including 36 combined with liver cirrhosis and 13 without liver cirrhosis;positive and negative AFPs were respectively detected in 34 and 15 patients.There was no statistically significant difference in gender,age,Child-Pugh score of liver function,liver cirrhosis and comparison of AFP between patients with and without sequential TACE (x2 =0.784,t =-0.512,x2 =0.013,2.844,P＞0.05).② Tumor recurrence:the 6-month and 1-year tumor recurrence rates after hepatectomy were respectively 0,28.57％ in 7 positive MVI patients with sequential TACE and 42.86％,51.02％ in 49 positive MVI patients without sequential TACE,showing a statistically significant difference in 6-month tumor recurrence rate (x2=4.800,P＜0.05) and no statistically significant difference in 1-year tumor recurrence rate (x2 =1.236,P ＞ 0.05).(4) Effect of comprehensive treatment on the prognosis of patients with tumor recurrence:of 59 patients with tumor recurrence,37 didn't receive comprehensive treatment,34 of 37 died within 1-year postoperatively,including 30 deaths within 6-month postoperatively,and 1-year survival rate was 8.10％;22 received comprehensive treatment,including 4 deaths within 1-year postoperatively,and 1-year survival rate was 81.80％,showing a statistically significant difference in 1-year survival rate (x2=32.698,P＜0.05).Conclusions MVI is one of the important risk factors affecting HCC recurrence and metastasis after hepatectomy.The combined TACE after hepatectomy can reduce the HCC recurrence rate of MVI positive patients,and active comprehensive treatment after HCC recurrence can significantly prolong the survival time of patients.

To investigate the changes in the expression levels of scavenger receptor class B member 1 (SCARB 1) in the liver tissues before and after liver xenotransplantation and analyze the relationship between the variations in the SCARB1 expression and coagulation regulating dysfunction in the recipients.Methods The Wuzhishan miniature pig with α-1,3-galactosyltransferase gene-knockout(GTKO) was utilized as the donor and Macaca thibetana was chosen as the recipient.Heterotopic auxiliary liver xenotransplantation models were established.The liver tissue specimen was collected before and after liver xenotransplantation.Primary hepatocytes were extracted from the pig using collagenase digestion method.Human peripheral blood mononuclear cells were obtained by immunomagnetic bead sorting.These two types of cells were co-cultured and supplemented with human plasma to establish cell models with coagulation regulating dysfunction following liver xenotransplantation.Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were performed to quantitatively measure and statistically compared the expression levels of messenger ribonucleic acid (mRNA) and protein of SCARB1 in the tissue and cell samples.At the cellular level,the expression of SCARB 1 was interfered by lentiviral vector.The coagulation time was detected to validate the effect upon coagulation function.Results The expression levels of SCARB1 mRNA and protein were significantly down-regulated after liver xenotransplantation (both P＜0.05).In the cell models,the expression levels of SCARB1 mRNA and protein in the porcine hepatocytes co-cultured with human monocytes were significantly down-regulated compared with those in porcine hepatocytes without intervention (both P＜0.05).Compared with the non-intervention group,the coagulation time was significantly prolonged after the expression of SCARB1 was interfered by lentiviral vector (P＜0.05).Conclusions The down-regulated expression of SCARB1 in the liver graft is one of the main causes of mediating coagulation regulating dysfunction.Intervention of SCARB1 expression contributes to resolve the coagulation regulating dysfunction in the recipients after liver xenotransplantation.

Objective To investigate the application prospect of the most extensive genome engineering pig internationally in preclinical xenotransplantation. Methods Porcine endogenous retrovirus (PERV) knockout combined with 3 major heterologous antigen gene knockouts and 9 humanized genes for inhibition of complement activation, regulation of coagulation disorders, anti-inflammatory and anti-phagocytosis were transferred into a pig (PERV-KO/3-KO/9-TG) as a donor, and the heart, liver and kidney were obtained and transplanted to 3 Rhesus macaque recipients respectively to establish a preclinical research model of pig-to-Rhesus macaque xenotransplantation. The functional status of xenografts after blood flow reconstruction was observed and the survival of recipients was summarized. The hemodynamics of xenografts were monitored. The change of hematological indexes of each recipient was compared. The histopathological manifestation of xenografts was observed. Results After the blood flow was reconstructed, all xenografts showed ruddy color, soft texture and good perfusion. The transplant heart, liver and kidney showed full arterial and venous blood flow and good perfusion at 1 d after operation. The postoperative survival time of heart, liver, and kidney transplant recipients was 7, 26, and 1 d, respectively. The levels of creatine kinase, creatine kinase isoenzyme, and lactate dehydrogenase increased in heart transplant recipient at 1 d after operation, and gradually recovered to near normal levels at 6 d after operation. All indexes increased sharply at 7 d after operation. The level of aspartate aminotransferase increased in liver transplant recipients at 2 d after operation, and the alanine aminotransferase basically returned to normal at 10 d after operation, but the total bilirubin continued to increase. Both aspartate aminotransferase and alanine aminotransferase increased at 12 d after operation, and reached a peak at 15 d after operation. The kidney transplant recipient developed mild proteinuria at 1 d after operation, and died of sudden severe arrhythmia. Histopathology showed that the tissue structure of cardiac and renal xenografts was close to normal, and liver xenografts presented with patchy necrosis, the liver tissue structure was disordered, accompanied by inflammatory damage, interstitial hemorrhage and thrombotic microangiopathy. Conclusions PERV-KO/3-KO/9-TG pig shows advantages in overcoming hyperacute rejection, mitigating humoral rejection and coagulation dysregulation. However, whether it can be used as potential donor for clinical xenotransplantation needs further evaluation.

【Objective】 To compare the short-term clinical effects of oblique lateral interbody fusion （OLIF） and transforaminal lumbar interbody fusion （TLIF） for treating single-segment lumbar spondylolisthesis. 【Methods】 We retrospectively analyzed the data of 68 patients with single-segment degenerative lumbar spondylolisthesis from January 2019 to February 2020. According to different surgical methods， the patients were divided into OLIF+ anterior screw fixation group （33 cases） and TLIF + posterior pedicle screw fixation group （35 cases）. The operation time， intraoperative blood loss， postoperative drainage， postoperative hospital stay and complication rate were compared between the two groups of patients. The disc height （DH）， lumbar lordosis （LL）， fused segmental lordosis （FSL）， foraminal height （FH）， and spondylolisthesis angle （SA） were measured before and after surgery and during follow-up. The visual analogue scale （VAS） of waist pain and the Oswestry disability index （ODI） were used to evaluate the short-term clinical efficacy. 【Results】 The operation time， intraoperative blood loss， postoperative drainage， and postoperative hospital stay were less in OLIF group than in TLIF group （all P<0.05）. There was no statistically significant difference in VAS or ODI scores between the two groups at the last follow-up after surgery （both P>0.05）. The two groups had statistically significant differences in DH and FH after surgery （P<0.05）， but no significant difference in postoperative LL， FSL or SA （all P>0.05）. There were six （18.2%） and five （14.3%） cases of complications in OLIF group and TLIF group， respectively， with no significant difference （P>0.05）. 【Conclusion】 OLIF and TLIF are equally safe and effective in treating single-segment lumbar spondylolisthesis. However， OLIF combined with anterior screw fixation has the advantages of less surgical trauma， less blood loss， shorter operation time， reduced postoperative hospital stay and shorter recovery time. Therefore， it is a more minimally invasive surgical option.

Objective: To explore the therapeutic feasibility of uterus transplantation for uterine infertility. Methods: Retrospective analysis was performed for the diagnosis, treatment and pregnancy course of the first domestic case of uterus transplantation and the relevant literature reviewed. The recipient was a 22-year-old woman with a congenital absence of uterus and vagina. Previously she underwent vaginal reconstruction and the donor was her mother. The specific procedures included donor/recipient screening, ethical argumentation, assisted reproductive technology of obtaining frozen embryos, Vinci robot-assisted uterine procurement, orthotopic replacement & fixation of retrieved uterus, revascularization; immunoregulation & monitoring of transplanted uterine recipient, assisted reproductive technology after transplantation and gestational management. Results: The durations of donor and recipient surgeries were 360 and 530 min respectively. No complications of recipient or donor occurred during the perioperative period. First menstruation occurred at 40 days post-transplantation and regularly thereafter. Pregnancy occurred after embryo transfer at 31 months post-transplantation. No rejection episodes occurred after transplantation or during gestation. Caesarean delivery occurred near gestational week 34. The boy weighed 2000 grams at birth and the mother remained well. Conclusions In conjunctions with literature review, uterine infertility may be treated by modified uterus transplantation. And a new path is paved for healthy pregnancy of women with uterine infertility.