Transplantation of human matrix metalloproteinase-1 gene-modified bone marrow-derived mesenchymal stem cell attenuates CCL4-induced liver fibrosis in rats.

It has been reported that bone marrow-derived mesenchymal stem cells (BMSCs) alleviated liver fibrosis. We investigated whether BMSCs transfected with human matrix metalloproteinase 1 (BMSCs/MMP1) would improve their therapeutic effect in liver fibrosis induced by carbon tetrachloride (CCl4) in rats. BMSCs were transfected with an adenovirus carrying enhanced green fluorescence protein (GFP) and human MMP1 gene. BMSCs or BMSCs/MMP1 were directly injected into fibrotic rats via the tail vein. GFP-labeled cells appeared in the fibrotic liver after BMSC transplantation. The expression of BMSCs/MMP1 elevated levels of MMP1 in vitro. Although BMSC administration reduced liver fibrosis, transplantation of BMSCs/MMP1 enhanced the reduction of liver fibrosis to a higher level. Treatment with BMSCs/MMP1 not only decreased collagen content but also suppressed activation of hepatic stellate cells (HSCs) in fibrotic liver, which led to subsequent improvement of both liver injury and fibrosis. Treatment with BMSCs/MMP1 resulted in an improved therapeutic effect compared with BMSCs alone, which is probably because of the sustainably expressed MMP1 level in the liver. BMSCs/MMP1 transplantation not only improved biochemical parameters but also attenuated progression of liver fibrosis, suggesting that BMSCs may be a potential cell source in preventing liver fibrosis and MMP1 gene may enhance the anti-fibrotic effect of BMSCs.

Polarization Spectroscopy of Defect-Based Single Photon Sources in ZnO.

Point defects in wide bandgap semiconductors are promising candidates for future applications that necessitate quantum light sources. Recently, defect-based single photon sources have been observed in ZnO that are very bright and remain photoactive from 4.5 K to room temperature. Despite several investigations, the structure and electronic states of these emitters remain unknown. In this work, we establish a procedure to distinguish a Z dipole from an XY dipole when studying quantum emitters that are randomly oriented. Our cryogenic and room temperature polarization measurements collectively establish that these unidentified ZnO quantum emitters have a Z dipole. We show that the associated absorption and emission dipoles are parallel within experimental uncertainty for all 32 individuals studied. Additionally, we apply group theory and find that, assuming the defect symmetry belongs to a point group relevant to the ZnO wurtzite lattice, the ground and excited states are orbital singlets. These results are a significant step in identifying the structure and electronic states of defect-based single photon sources in ZnO.

Repeated pancreatitis-induced splenic vein thrombosis leads to intractable gastric variceal bleeding: A case report and review.

Gastric varices (GV) are one of the most common complications for patients with portal hypertension. Currently, histoacryl injection is recommended as the initial treatment for bleeding of GV, and this injection has been confirmed to be highly effective for most patients in many studies. However, this treatment might be ineffective for some types of GV, such as splenic vein thrombosis-related localized portal hypertension (also called left-sided, sinistral, or regional portal hypertension). Herein, we report a case of repeated pancreatitis-induced complete splenic vein thrombosis that led to intractable gastric variceal bleeding, which was treated by splenectomy. We present detailed radiological and pathological data and blood rheology analysis (the splenic artery - after a short gastric vein or stomach vein - gastric coronary vein - portal vein). The pathophysiology can be explained by the abnormal direction of blood flow in this patient. To our knowledge, this is the first reported case for which detailed pathology and blood rheology data are available.

Contrast enhanced computed tomography and reconstruction of hepatic vascular system for transjugular intrahepatic portal systemic shunt puncture path planning.

AIM: To describe a method for the transjugular intrahepatic portal systemic shunt (TIPS) placement performed with the aid of contrast-enhanced computed tomography (CECT) and three-dimensional reconstructed vascular images (3D RVIs), and to assess its safety and effectiveness. METHODS: Four hundred and ninety patients were treated with TIPS between January 2005 and December 2012. All patients underwent liver CECT and reconstruction of 3D RVIs of the right hepatic vein to portal vein (PV) prior to the operation. The 3D RVIs were carefully reviewed to plan the puncture path from the start to target points for needle pass through the PV in the TIPS procedure. RESULTS: The improved TIPS procedure was successful in 483 (98.6%) of the 490 patients. The number of punctures attempted was one in 294 (60%) patients, 2 to 3 in 147 (30%) patients, 4 to 6 in 25 (5.1%) patients and more than 6 in 17 (3.5%) patients. Seven patients failed. Of the 490 patients, 12 had punctures into the artery, 15 into the bile duct, eight into the gallbladder, and 18 through the liver capsule. Analysis of the portograms from the 483 successful cases indicated that the puncture points were all located distally to the PV bifurcation on anteroposterior images, while the points were located proximally to the bifurcation in the three cases with intraabdominal bleeding. The complications included three cases of bleeding, of whom one died and two needed surgery. CONCLUSION: Use of CECT and 3D RVIs to plan the puncture path for TIPS procedure is safe, simple and effective for clinical use.

Hepatoprotective effect of juglone on dimethylnitrosamine-induced liver fibrosis and its effect on hepatic antioxidant defence and the expression levels of α-SMA and collagen III.

The present study aimed to investigate the antifibrotic effects of juglone on dimethylnitrosamine (DMN)­induced fibrosis in rats. Juglone, which is a quinone, significantly decreased DMN­induced rat hepatic fibrosis, which was associated with increased superoxide dismutase (SOD) activity, decreased oxidative stress and reduced levels of α­smooth muscle actin (α­SMA) and collagen (Col) III in the liver. Serum levels of alanine aminotransferase, aspartate aminotransferase, hyaluronic acid, laminin, type III precollagen and type IV collagen were significantly reduced by treatment with juglone. Liver fibrosis was induced in male Sprague­Dawley rats by subcutaneous injections of DMN solution and hepatic fibrosis was assessed using Massons trichome staining. The expression levels of α­SMA and Col III were determined using immunohistochemical techniques. The activities of SOD and malondialdehyde in liver homogenates were also determined. The results suggested that juglone augmented the antioxidative capability of the liver, possibly by stimulating the activity of SOD, which promoted the inactivation of hepatic stellate cells (HSCs) and decreased the accumulation of extracellular matrix collagen in the liver, thereby alleviating hepatic fibrosis. Silymarin was used as a positive control for liver fibrosis protection. It was hypothesized that juglone alleviates or mitigates oxidative stress­mediated hepatic fibrosis by upregulating the expression of peroxisome proliferator­activated receptor γ and inhibiting the activation of HSC.

[The agreement and clinical value of hepatic vein pressure gradient and portal vein pressure in patients with portal hypertension].

OBJECTIVE: To evaluate the agreement and correlation between hepatic vein pressure gradient (HVPG) and portal vein pressure (PVP) in patients with portal hypertension,and explore their clinical value. METHODS: A total of 46 patients with portal hypertension were directly measured the free hepatic pressure, wedged hepatic pressure, portal vein pressure before and after TIPS therapy. The agreement and correlation of HVPG and PVP were analyzed, and explore their clinical value. RESULTS: There is no significant agreement or correlation between HVPG and PVP in 5 patients, whose third hilar have large communicating branches between portal vein and Inferior vena cava, or with obvious umbilical vein opened. The HVPGs were significantly agreed with portal vein pressure in other 41 patients. There is no significant difference of HVPG or PVP between earlyTIPS and not early-TIPS groups. In addition, the portal vein pressures after TIPS were significantly decreased compared with that before TIPS. CONCLUSION: The HVPG can well show the PVP except these with obvious communicating branches between portal vein and Inferior vena cava in third hilar, and TIPS can effectively decrease the portal vein pressure in patients with portal hypertension.

Portal hypertension caused by right common iliac artery-superior mesenteric vein fistula.

Portal hypertension caused by arterio-portal fistula between the right common iliac artery and superior mesenteric vein has not been reported. Here, we report such a case in a 35-year-old male without any history of liver disease or abdominal trauma. This case was confirmed by abdominal aorta angiography and three-dimensional angiography, and was successfully treated by a covered stent implantation in the right common iliac artery to block the fistula.

Overexpression of p28GANK accelerates the metastasis of oesophageal squamous cell carcinoma.

The present study aimed to evaluate the effects of p28GANK expression on the metastasis of oesophageal squamous cell carcinoma (ESCC) tissues and to investigate its roles in the metastasis of highly invasive and non­invasive ESCC cell lines. Quantitative polymerase chain reaction (qPCR) and immunohistochemical analyses were performed to assess p28GANK mRNA and protein expression in ESCC tissues and to analyse its significance in ESCC metastasis. qPCR and western blot analyses were used to detect p28GANK mRNA and protein expression in highly invasive and non­invasive cell lines. Subsequently, lentivirus­mediated p28GANK short interfering RNA (siRNA) was transfected into highly invasive ESCC cells, and Transwell assays were performed to analyse the effects of p28GANK knockdown on their migration and invasion. The mean expression levels of p28GANK mRNA in the ESCC tissues of patients with metastasis were significantly higher than those in the ESCC specimens from patients without metastasis. p28GANK expression in ESCC tissues was correlated with T­stage, lymph node metastasis and lymphatic invasion. The mRNA and protein expression levels of p28GANK were significantly higher in highly invasive cell lines compared with those of matched, non­invasive cell lines. Lentivirus­mediated siRNA knockdown of p28GANK markedly decreased p28GANK expression in EC109­P and EC9706­P cells and supressed the metastasis of ESCC cells in vitro. In conclusion, p28GANK expression was increased in metastatic ESCC tissues and cells, and p28GANK knockdown decreased the metastatic ability of ESCC cells. These results suggested that p28GANK may be a potential therapeutic marker for ESCC metastasis.

Retrospective analysis of venograms of hepatic and portal veins: clinical implications for transjugular intrahepatic portosystemic shunt placement.

AIMS: This study was aimed to provide safety guidance of needle passes into the portal vein during transjugular intrahepatic portosystemic shunt (TIPS) placement. METHODOLOGY: On anteroposterior venograms, the orifice of right hepatic trunk (RHT), furcation position, and branching of portal trunk in 128 patients underwent TIPS were mapped in relation to the vertebrae and intervertebral space. Impact of clinical factors on these parameters was determined. RESULTS: The orifices of RHTs were all above the furcation position of portal trunk, RHTs were posterior and superior to portal branch. Of the 128 patients, 84.4% had the orifice positioned between 9th and 10th thoracic (T) vertebrae, 58.6% positioned to T10. Portal trunks were furcated between T11 and T12 in 80.5% patients. Portal trunks were bifurcated at right hepatic portal in 91.4% patients into left and right veins, and trifurcated into right posterior, right anterior and left branches in 8.6% patients. Statistical analysis indicated that these parameters were not affected by cause of disease, gender, age and Child-Pugh score. CONCLUSIONS: The puncture site for portal vein located at or beyond imaged trunk furcation would be safe for most of the patients.

[Therapeutic effect of BMSCs with over-expressed MMP1 on liver fibrosis].

OBJECTIVE: To investigate the function of bone marrow mesenchymal stem cells (BMSCs) with over-expressed matrix metalloproteinase 1 (MMP1) on liver fibrosis. METHODS: Fifty SD male rats were randomly divided into 4 groups: recombinant adenovirus Adhuman MMP-1(hMMP-1)-enhanced green fluorescent protein (EGFP) transfected BMSCs group (Group A, n=10), Ad-EGFP transfected BMSCs group (Group B, n=10), liver fibrosis group (Group C, n=15), and a normal group (Group D, n=15). The liver fibrosis model was formed by subcutaneous injection of the mixed liquor of carbon tetrachloride (CCL4) and vegetable oil. After 10 weeks, the model of liver fibrosis was formed. Group A and B were administered the transfected BMSCs via the tail veins, while Group C and D were administered normal saline. After 3 weeks, the rats were sacrificed. The body weight, liver weight, liver function, liver fibrosis indexes and liver pathological changes were tested. RESULTS: Compared with the control group, the rats administered BMSCs with over-expressed MMP1 showed a significant improvement in the body weight, liver weight and plasma albumin (ALB) (P<0.05), and a significant reduction in the plasma alanine aminotransferase, total bilirubin, hyaluronic acid, laminin and procollagen III (P<0.05). Hematoxylin-eosin staining confirmed that the degree of liver fibrosis was significantly ameliorated under average visual fields (P<0.05). CONCLUSION: The repair ability of BMSCs on liver fibrosis can be enhanced by over-expression of hMMP-1.

[Significance of SAMD9 expression in esophageal squamous cell carcinoma].

OBJECTIVE: To analyze the significance of sterile alpha motif domain-containing 9 (SAMD9) expression in esophageal squamous cell carcinoma (ESCC). METHODS: Immunohistochemical staining was performed to detect the expression of SAMD9 in 72 primary ESCC and matched adjacent cancer-free tissues and analyze the significance of SAMD9 expression in ESCC tissues. In addition, Western blotting was used to detect the expression of SAMD9 in primary ESCC tissues which were taken from 3 metastatic and 3 non-metastatic patients during surgery. RESULTS: The expression of SAMD9 in the ESCC tissues had no statistical difference from that in the matched adjacent cancer-free tissues. SAMD9 expression was significantly correlated with lymphatic invasion and metastasis in these patients (P<0.05), but not with age, gender, tumor differentiation and T stage (P>0.05). Western blotting showed that SAMD9 expression in primary ESCC tissues was significantly higher in the patients with metastasis than in the ones without metastasis(P<0.01). CONCLUSION: Over-expression of SAMD9 is correlated with the metastasis of ESCC, indicating that it might play an important role in the metastasis of ESCC.

Clinical effects and complications of TIPS for portal hypertension due to cirrhosis: a single center.

AIM: To determine the clinical effects and complications of transjugular intrahepatic portosystemic shunt (TIPS) for portal hypertension due to cirrhosis. METHODS: Two hundred and eighty patients with portal hypertension due to cirrhosis who underwent TIPS were retrospectively evaluated. Portal trunk pressure was measured before and after surgery. The changes in hemodynamics and the condition of the stent were assessed by ultrasound and the esophageal and fundic veins observed endoscopically. RESULTS: The success rate of TIPS was 99.3%. The portal trunk pressure was 26.8 ± 3.6 cmH2O after surgery and 46.5 ± 3.4 cmH2O before surgery (P < 0.01). The velocity of blood flow in the portal vein increased. The internal diameters of the portal and splenic veins were reduced. The short-term hemostasis rate was 100%. Esophageal varices disappeared completely in 68% of patients and were obviously reduced in 32%. Varices of the stomach fundus disappeared completely in 80% and were obviously reduced in 20% of patients. Ascites disappeared in 62%, were markedly reduced in 24%, but were still apparent in 14% of patients. The total effective rate of ascites reduction was 86%. Hydrothorax completely disappeared in 100% of patients. The incidence of post-operative stent stenosis was 24% at 12 mo and 34% at 24 mo. The incidence of post-operative hepatic encephalopathy was 12% at 3 mo, 17% at 6 mo and 19% at 12 mo. The incidence of post-operative recurrent hemorrhage was 9% at 12 mo, 19% at 24 mo and 35% at 36 mo. The cumulative survival rate was 86% at 12 mo, 81% at 24 mo, 75% at 36 mo, 57% at 48 mo and 45% at 60 mo. CONCLUSION: TIPS can effectively lower portal hypertension due to cirrhosis. It is significantly effective for hemorrhage of the digestive tract due to rupture of esophageal and fundic veins and for ascites and hydrothorax caused by portal hypertension.

Quantification of serum hepatitis B surface antigen in predicting the response of pegylated interferon alfa-2a in HBeAg-positive chronic hepatitis B with prior lamivudine exposure.

AIMS: Majority of previous studies of pegylated interferon α-2a (PegIFNα-2a) forced on naïve chronic hepatitis B (CHB) patients, and the data of PegIFNα-2a in therapy of patients with prior exposure to nucleos(t)ide analogues is rare. This study aimed to investigate the predictive role of serum quantitative hepatitis B surface antigen (HBsAg) in predicting sustained response of PegIFNα-2a in HBeAg-positive CHB patients with prior lamivudine exposure. METHODS: Forty-six patients with prior lamivudine exposure received PegIFNα-2a for 12 months and followed-up for 6 months. The clinical features of responders and non-responders were compared, and the predictive role of quantitative HBsAg in predicting responders at the end of follow-up was evaluated. Responders were defined as an ALT normalization, HBeAg seroconversion and sustained virological response at the end of follow-up. RESULTS: In this cohort, only 26.1% (12/46) patients were responders. The baseline characteristics of the responders and non-responders were similar; however, the rates of ALT normalization, HBV DNA undetectability and HBeAg seroconversion were all significantly higher in responders than that in non-responders. During the treatment and follow-up, the HBsAg levels were all significantly lower in responders than that in non-responders. In predicting reponders, the serum HBsAg cutoff of 6000 IU/mL at months 6 had a positive predictive value of 73.3 and a negative predictive value of 96.8%, and with an area under the receiver operating characteristic curve of 0.869. CONCLUSION: The responders toward PegIFNα-2a in CHB patients with prior lamivudine exposure is not high, and serum HBsAg <6000 IU/Ml at months 6 of on-treatment had a high value to predict long-term outcomes of treatment.

[Recombinant adenovirus Ad-human matrix metalloproteinase 1 transfecting bone marrow mesenchymal stem cells of rats in vitro].

OBJECTIVE: To transfect bone marrow mesenchymal stem cells (BMSCs) of rats by recombinant adenovirus Ad-human matrix metalloproteinase 1 (hMMP-1) in vitro so as to lay the experimental foundation for the treatment of liver fibrosis with a combination of BMSCs and hMMP-1 gene transplantation. METHODS: BMSCs were isolated from bone marrow of 2-3 weeks old Sprague Dawley rats by whole bone marrow adherence method and identified, then transfected by recombinant adenovirus Ad-hMMP-1 carrying enhanced green fluorescent protein (EGFP) marker in vitro. The green fluorescent expression was observed by fluorescence microscope and the transfection efficiency was detected by flow cytometry to determine the optimum multiplicity of infection (MOI). BMSCs at passage 3 were divided into 3 groups: untransfected BMSCs group (group A), Ad-EGFP transfected BMSCs group (group B), and Ad-hMMP-1-EGFP transfected BMSCs group (group C); the gene and intracellular protein of hMMP-1 were detected by RT-PCR and Western blot; the ELISA assay was used to detect the supernatant protein expression, and the hMMP-1 activity was measured by fluorescent quantification kit. RESULTS: The green fluorescent was observed in BMSCs transfected by recombinant adenovirus at 24 hours after transfection; the fluorescence intensity was highest at 72 hours; and the optimum MOI was 200. The cells of 3 groups entered the logarithmic growth phase on the 3rd day and reached plateau phase on the 6th day by MTT assay; no significant difference was found in the cell proliferation rate among 3 groups (P > 0.05). RT-PCR, Western blot, and ELISA assay showed high expressions of the hMMP-1 gene and protein in group C, but no expression in groups A and B. The hMMP-1 activity was 1.24 nmol/(mg.min) in group C, but hMMP-1 activity was not detectable in groups A and B. CONCLUSION: The exogenous hMMP-1 gene is successfully transfected into BMSCs of rats via recombinant adenovirus and can highly express, which lays the experimental foundation for the treatment of liver fibrosis with a combination of BMSCs and hMMP-1 gene transplantation.

Meta-analysis of the efficacy of sorafenib for hepatocellular carcinoma.

PURPOSE: By carrying out a meta-analysis of randomized controlled trials that compared sorafenib or combined chemotherapy with placebo or combined chemotherapy, the effectiveness of sorafenib in hepatocellular carcinoma was evaluated in the present study, which also provided clinical practice guidelines of evidence-based-medicine. METHODS: We reviewed PubMed citations concerning sorafenib treating hepatocellular carcinoma in randomized controlled trials from Jan 2000 to July 2012. All the literature was extracted by Cochrane systematic reviews and underwent meta-analysis with RewMan 5.0 software. RESULTS: Finally, four papers documenting randomized controlled studies were included. Compared with controls, sorafenib was shown to significantly increase overall survival (OS), time to progression (TTP), and disease control rates (DCR), but not the time to symptom progression (TTSP) in hepatocellular carcinoma patients. The incidence of grade-III/IV adverse reactions, including hand- foot-skin reactions, diarrhea, hypertension and skin rash or desquamation, in sorafenib treatment group was higher than that in controls. However, there was no significant difference in the incidence of hypodynamia between the two groups. CONCLUSIONS: Sorafenib exerts significant curative effects in hepatocellular carcinoma.

Sustained efficacy of adefovir add-on therapy in chronic hepatitis B patient with a poor virological response to peginterferon alfa.

BACKGROUND: Currently, there is no consensus on the recommendation of peginterferon alfa (pegIFNα) to chronic hepatitis B (CHB) patients with poor viral response (EVR). This study aimed to assess the sustained curative efficacy of adefovir (ADV) add-on therapy in optimizing pegIFNα monotherapy. METHODS: A total of 85 hepatitis B e antigen (HBeAg)-positive CHB patients with poor virological response at month 6 after starting pegIFNα-2a were enrolled, and received either pegIFNα-2a continuing monotherapy (group A, n = 51) or add-on therapy with ADV (group B, n = 34). The treatment duration for all patients was 6 months, and the sustained responses after the end of treatment were evaluated between two groups. RESULTS: The baseline characteristics were comparable between two groups. At months 6 after treatment completion, the sustained virological response (SVR) rates were 31.4% and 73.5%, the sustained biochemical response (SBR) rates were 39.2% and 85.3% in group A and group B respectively, and the difference in either SVR or SBR was statistically significant (both p < 0.001). As compared to patients in group A, significantly more patients in group B obtained HBeAg loss (19.6% vs. 55.9%, p = 0.001) and seroconversion (13.7% vs. 41.2%, p = 0.004). CONCLUSION: ADV add-on therapy could significantly improve and sustain the curative efficacy of CHB patient with poor virological response to pegIFNα-2a monotherapy, but further large well-designed randomized controlled trials are needed to confirm our findings.

Primary intestinal NK/T cell lymphoma: a clinicopathologic study of 25 Chinese cases.

BACKGROUND: Primary intestinal NK/T cell lymphoma is extremely rare and early diagnosis is frequently difficult. The aim of this study is to investigate the clinicopathological findings, immunophenotype, and T cell receptor (TCR) γ gene rearrangement of primary intestinal NK/T cell lymphomas in 25 Chinese cases. METHODS: Clinical data of the 25 cases were analyzed. Immunohistochemistry for immunophenotype, in situ hybridization for EBER, and polymerase chain reaction for TCR γ gene rearrangement were investigated. Survival curves according to clinical characteristics were analyzed. RESULTS: The median age was 33 years and the median survival was 7 months. The common symptoms consisted of abdominal pain, fever, marasmus, diarrhea, and hematochezia. Endoscopically, the tumors were mainly featured by focal, multifocal or diffuse irregular ulcers, which most frequently emerged in the ascending colon. Histologically, the tumors were characterized by the proliferation of pleomorphic atypical lymphoid cells (ALCs), necrosis, lympho-epithelial lesions, and mixed inflammatory infiltration. The positive frequency of CD3ε was 88.2%, of CD56 was 84%, granzyme B was 90%, and EBER was 84.2%. A total of 12 out of 14 cases (85.7%) highly expressed Ki67. The negative prognostic factors for survival were Ann Arbor stage IIIE or IVE (P = 0.039) and more than one extranodal site of disease (P = 0.019). CONCLUSION: Primary intestinal NK/T cell lymphomas most frequently favor young people and have a poor prognosis. Due to the nonspecific clinical and endoscopic findings, it is difficult to distinguish intestinal NK/T cell lymphomas from inflammatory and infectious disorders. Histopathology, immunophenotype, and DNA study play key roles in differential diagnosis.