Effect of postoperative adjuvant chemotherapy on prognosis of patients with intrahepatic cholangiocarcinoma:a multicenter retrospective study / 中华外科杂志

Objectives: To examine the influence of adjuvant chemotherapy after radical resection on the survival of patients with intrahepatic cholangiocarcinoma(ICC) and to identify patients who may benefit from it. Methods: The clinical and pathological data of 654 patients with ICC diagnosed by postoperative pathology from December 2011 to December 2017 at 13 hospitals in China were collected retrospectively. According to the inclusion and exclusion criteria,455 patients were included in this study,including 69 patients (15.2%) who received adjuvant chemotherapy and 386 patients (84.8%) who did not receive adjuvant chemotherapy. There were 278 males and 177 females,with age of 59 (16) years (M(IQR))(range:23 to 88 years). Propensity score matching (PSM) method was used to balance the difference between adjuvant chemotherapy group and non-adjuvant chemotherapy group. Kaplan-Meier method was used to plot the survival curve,the Log-rank test was used to compare the difference of overall survival(OS) and recurrence free survival(RFS)between the two groups. Univariate analysis was used to determine prognostic factors for OS. Multivariate Cox proportional hazards models were then performed for prognostic factors with P<0.10 to identify potential independent risk factors. The study population were stratified by included study variables and the AJCC staging system,and a subgroup analysis was performed using the Kaplan-Meier method to explore the potential benefit subgroup population of adjuvant chemotherapy. Results: After 1∶1 PSM matching,69 patients were obtained in each group. There was no significant difference in baseline data between the two groups (all P>0.05). After PSM,Cox multivariate analysis showed that lymph node metastasis (HR=3.06,95%CI:1.52 to 6.16,P=0.039),width of resection margin (HR=0.56,95%CI:0.32 to 0.99,P=0.044) and adjuvant chemotherapy (HR=0.51,95%CI:0.29 to 0.91,P=0.022) were independent prognostic factors for OS. Kaplan-Meier analysis showed that the median OS time of adjuvant chemotherapy group was significantly longer than that of non-adjuvant chemotherapy group (P<0.05). There was no significant difference in RFS time between the adjuvant chemotherapy group and the non-adjuvant chemotherapy group (P>0.05). Subgroup analysis showed that,the OS of female patients,without HBV infection,carcinoembryonic antigen<9.6 μg/L,CA19-9≥200 U/ml,intraoperative bleeding<400 ml,tumor diameter>5 cm,microvascular invasion negative,without lymph node metastasis,and AJCC stage Ⅲ patients could benefit from adjuvant chemotherapy (all P<0.05). Conclusion: Adjuvant chemotherapy can prolong the OS of patients with ICC after radical resection,and patients with tumor diameter>5 cm,without lymph node metastasis,AJCC stage Ⅲ,and microvascular invasion negative are more likely to benefit from adjuvant chemotherapy.

Focus problems in diagnosis and treatment of recurrent hepatocellular carcinom / 中国实用外科杂志

Recurrent hepatocellular carcinoma(HCC) is acommon problem in the treatment of HCC patients. At present,there is a lack of staging and treatment system for recurrenthepatocellular carcinoma,and a lack of high-level clinicalresearch. Therefore,there is much controversy in the treatmentof recurrent hepatocellular carcinoma. The controversy ismainly focused on whether it is proper to take aggressivelyrepeated surgical resection or liver transplantation. For recurrent tumors of polyclonal origin,clinical treatmentdecisions are basically based on the staging and treatmentsystem of primary hepatocellular carcinoma,and the prognosisis not inferior to that of primary tumors. For recurrent tumorsof monoclonal origin,it tends to be less invasive,such asinterventional,ablation,radiotherapy or targeted drugs. Thetreatment strategy for hepatocellular carcinoma must considermany factors, such as trauma,safety,effectiveness and cost. Itis similar to and still a little different from that of primaryhepatocellular carcinoma. Therefore,the treatment of recurrenthepatocellular carcinoma cannot fully refer to the treatmentsystem of primary hepatocellular carcinoma.

Anti-inflammatory effect of external use of escin on cutaneous inflammation: possible involvement of glucocorticoids receptor / 中国天然药物

Escin, as an internally applied anti-inflammatory agent, has been widely used in the treatment of inflammation and edema resulting from trauma or operation in the clinic. However, the effect of its external use on cutaneous inflammation and edema remains unexplored. In the present study, the anti-inflammatory and anti-edematous effects of external use of escin were studied in carrageenan-induced paw edema and histamine-induced capillary permeability in rats, paraxylene-induced ear swelling in mice, and cotton pellet-induced granuloma in rats. Effects of external use of escin gel on prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) were determined by ELISA. The anti-inflammatory mechanism was explored by detecting the expression of glucocorticoid receptor (GR) with Western blotting and Real-time PCR analyses, with further exploration of nuclear factor-κB (NF-κB), p38 mitogen-activated protein kinase (P38MAPK) and activator protein-1 (AP-1) expressions. We demonstrated that external use of escin showed significant anti-inflammatory effects on acute and chronic inflammation in different animal models and its anti-inflammatory effects might be related to down-regulation of PGE2, TNF-α, and IL-1β. The results also showed that escin exerted its anti-inflammatory effects by promoting the expression of GR, with the possible mechanism being inhibition of the expressions of GR-related signaling molecules such as NF-κB and AP-1.

Anti-inflammatory effect of external use of escin on cutaneous inflammation: possible involvement of glucocorticoids receptor / 中国天然药物

Escin, as an internally applied anti-inflammatory agent, has been widely used in the treatment of inflammation and edema resulting from trauma or operation in the clinic. However, the effect of its external use on cutaneous inflammation and edema remains unexplored. In the present study, the anti-inflammatory and anti-edematous effects of external use of escin were studied in carrageenan-induced paw edema and histamine-induced capillary permeability in rats, paraxylene-induced ear swelling in mice, and cotton pellet-induced granuloma in rats. Effects of external use of escin gel on prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) were determined by ELISA. The anti-inflammatory mechanism was explored by detecting the expression of glucocorticoid receptor (GR) with Western blotting and Real-time PCR analyses, with further exploration of nuclear factor-κB (NF-κB), p38 mitogen-activated protein kinase (P38MAPK) and activator protein-1 (AP-1) expressions. We demonstrated that external use of escin showed significant anti-inflammatory effects on acute and chronic inflammation in different animal models and its anti-inflammatory effects might be related to down-regulation of PGE2, TNF-α, and IL-1β. The results also showed that escin exerted its anti-inflammatory effects by promoting the expression of GR, with the possible mechanism being inhibition of the expressions of GR-related signaling molecules such as NF-κB and AP-1.

Comparison of laparoscopic and open surgery in treatment of rectal cancer patients / 中国内镜杂志

Objective To compare the effect of laparoscopic and open surgery in treatment of rectal cancer. Methods 80 cases of patients with rectal cancer from May 2008 to May 2013 were selected, they were randomly divided into laparoscopy surgery group (n = 40) and open surgery group (n = 40), the operation time, intraoperative blood loss, length of incision, lymph node dissection, number for the first time, ventilation time, ambulation time, hospitalization time, cost of hospitalization, postoperative complications, treatment satisfaction of the two groups were statistically analyzed. Results The operation time of the laparoscopic surgery group was significantly longer (P < 0.05), the amount of bleeding was significantly less (P < 0.05), the incision length was significantly shorter (P < 0.05), the first time, ventilation time, ambulation time, hospitalization time were significantly shorter (P < 0.05), the hospitalization cost was significantly higher (P < 0.05), the rate of postoperative complications 15.0% (6/40) was significantly lower than the open surgery group 35.0% (14/40) (P < 0.05) 97.5% (39/40), the treatment satisfaction was significantly higher than the open surgery group 67.5% (27/40)(P < 0.05). Conclusion The effect of laparoscopic and open surgery in treatment of rectal cancer is better than open surgery.

Clinical practice guidelines for the treatment of primary liver cancer with integrative traditional Chinese and Western medicine / 中西医结合学报

Traditional Chinese medicine (TCM) is an important part of the treatment of primary liver cancer (PLC) in China; however, the current instructions for the integrative use of traditional Chinese and Western medicine for PLC are mostly based on expert opinion. There is no evidence-based guideline for clinical practice in this field. Therefore, the Shanghai Association of Chinese Integrative Medicine has established a multidisciplinary working group to develop this guideline, which focuses on the most important questions about the use of TCM during PLC treatment. This guideline was developed following the methodological process recommended by the World Health Organization Handbook for Guideline Development. Two rounds of questionnaire survey were performed to identify clinical questions; published evidence was searched; the Grading of Recommendations Assessment, Development and Evaluation approach was used to evaluate the body of evidence; and recommendations were formulated by combining the quality of evidence, patient preferences and values, and other risk factors. The guideline was written based on the Reporting Items for Practice Guidelines in Healthcare tool. This guideline contains 10 recommendations related to 8 questions, including recommendations for early treatment by TCM after surgery, TCM combined with transcatheter arterial chemoembolization for advanced PLC, TCM drugs for external use, and acupuncture and moxibustion therapy.

Effects of agarwood formation induced by Fusarium sp. A2 on distribution and community composition of endophytic fungi in leaves of Aquilaria sinensis / 中国中药杂志

The aim of the paper is to observe the distribution of the endophytic fungi in leaves of Aquilaria sinensis by using permanent paraffin-cut section, optical microscope photography and histochemistry. Total DNA was extracted with modified CTAB method and rDNA ITS regions of plant and endophytic fungi were amplified with eukaryotic universal primers. The rDNA ITS amplicon was characterized by RFLP analysis, sequencing of rDNA ITS library and phylogenetic analyses using PAUP by maximum parsimony. Fusarium sp. A2 was used to induce the formation of resinous in A. sinensis trees. As a result, endophytic fungi mainly distributed in spongy and phloem in leaf. Endophytic fungi distributed in the phloem in agarwood-producing tree and had a relatively high abundance. Phoma sp. and Collectrotrichum sp. were the absolute advantage species in the leaf tissues of non-resinous and agarwood-producing tree, respectively. Collectrotrichum sp. was the only fungal species detected both in the two types of A. sinensis with different levels of abundance. The culture-independent molecular method can be used to identify fungal species directly and rapidly from the plant tissues. Endophytic fungal communities in non-resinous and agarwood-producing A. sinensis leaf tissues were quite different.

Traditional herbal medicine in preventing recurrence after resection of small hepatocellular carcinoma: a multicenter randomized controlled trial / 中西医结合学报

<p><b>BACKGROUND</b>Disease recurrence is a main challenge in treatment of hepatocellular carcinoma (HCC). There is no generally accepted method for preventing recurrence of HCC after resection.</p><p><b>OBJECTIVE</b>To compare the efficacy of a traditional herbal medicine (THM) regimen and transarterial chemoembolization (TACE) in preventing recurrence in post-resection patients with small HCC.</p><p><b>DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS</b>This is a multicenter, open-label, randomized, controlled study, which was undertaken in five centers of China. A total of 379 patients who met the eligibility criteria and underwent randomization were enrolled in this trial. One hundred and eighty-eight patients were assigned to the THM group and received Cinobufacini injection and Jiedu Granule, and the other 191 patients were assigned to the TACE group and received one single course of TACE.</p><p><b>MAIN OUTCOME MEASURES</b>Primary outcome measures were the annual recurrence rate and the time to recurrence. Incidence of adverse events was regarded as the secondary outcome measure.</p><p><b>RESULTS</b>Among the 364 patients who were included in the intention-to-treat analysis, 67 patients of the THM group and 87 of the TACE group had recurrence, with a hazard ratio of 0.695 (P = 0.048). Median recurrence-free survival of the patients in the THM and TACE groups was 46.89 and 34.49 months, respectively. Recurrence rates at 1, 2 and 3 years were 17.7%, 33.0% and 43.5% for the THM group, and 28.8%, 42.5% and 54.0% for the TACE group, respectively (P = 0.026). Multivariate analysis indicated that the THM regimen had a big advantage for prolonging the recurrence-free survival. Adverse events were mild and abnormality of laboratory indices of the two groups were similar.</p><p><b>CONCLUSION</b>In comparison with TACE therapy, the THM regimen was associated with diminished risk of recurrence of small-sized HCC after resection, with comparable adverse events. TRIAL REGISTRTION IDENTIFIER: This trial was registered in the Chinese Clinical Trial Registry with the identifier ChiCTR-TRC-07000033.</p>

Long non-coding RNA influences radiosensitivity of colorectal carcinoma cell lines by regulating cyclin D1 expression / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To screen long non-coding RNA which influences radiosensitivity of colorectal carcinoma cell lines and investigate the mechanism.</p><p><b>METHODS</b>Under different doses of radiation, colony formation assay and single-hit multi-target model were conducted to draw dose-survival curve and SF2 value of colorectal carcinoma cell lines(RKO, Lovo) was calculated. High-throughput lncRNA/mRNA chips were used to screen lncRNA genes and protein coding genes with expression differences more than 2 folds between RKO, Lovo cell lines and RKO cell line receiving 2Gy radiation. The main action pathway was computed by Gene Ontology analysis combined with Pathway analysis in order to explore the mechanism which induces the effect of lncRNA on radiosensitivity of colorectal carcinoma cell lines. Further experiment on P53, P21, cyclin D1 expression contents of RKO cell line was confirmed by real-time RT-PCR.</p><p><b>RESULTS</b>Lovo(SF2=0.47) was more sensitivity to radiation than RKO(SF2=0.53) according to the outcome of colony formation assay. High-throughput lncRNA/mRNA chips identified a total of 268 lncRNA genes and 270 protein coding genes. Gene Ontology analysis showed that the expression of genes associated with cell cycle process were significantly different (38.6%). There was a significant relationship between expression of several lncRNAs and CCND1 gene. Real-time RT-PCR showed no significant differences of P53 and P21 expression in RKO and Lovo cell lines(P>0.05), while cyclin D1 expression of RKO cell line was higher than that of Lovo cell lines(P<0.05). After exposed to 2 Gy doses of radiation, there was an obvious decrease of cyclin D1 expression in RKO cell lines(P<0.05), while P53 and P21 expressions were not different(P>0.05).</p><p><b>CONCLUSION</b>The possible mechanism is that lncRNAs compose transcription compound to combine with CCND1 gene and influence radiosensitivity of colorectal carcinoma cell lines by regulating expression of cyclin D1, which is independent of P53-P21-cyclin D1 pathway.</p>

Hepatic vein occlusion with Satinsky clamp in hepatectomy for complicated liver tumor / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the application of an improved method of hepatic vein occlusion with Satinsky clamp when resecting the liver tumor involving second hepatic portal.</p><p><b>METHODS</b>From January 2003 to December 2010, there were totally 330 patients with liver tumor admitted, who underwent liver resection with Pringle maneuver plus hepatic vein occlusion with Satinsky clamp. Data regarding the intra-operative and post-operative course of the patients were analyzed. There were 245 male and 85 female patients, with a mean age of (50 ± 11) years. The diameter of tumor was (9 ± 6) cm. Among the 330 patients, there were 271 patients with viral hepatitis B, 215 patients with liver cirrhosis; 321 patients were in Child class A of liver function and 9 in class B. Pringle maneuver plus hepatic vein occlusion with Satinsky clamp was used to occlude the blood flow in the liver resection. The liver transection was performed with clamp-crushing technique.</p><p><b>RESULTS</b>Hepatic vein occlusion with Satinsky clamp was successful in all 330 patients. The operation time was (132 ± 29) minutes, while (7 ± 3) minutes for dissecting hepatic vein and (22 ± 7) minutes for inflow blood occlusion. The blood loss in operation was (480 ± 265) ml, with 20% of patients receiving blood transfusion. No patient had large hemorrhage and air embolism due to hepatic vein laceration. No patient died in the perioperative period. The complications included 31 patients of pleural effusion, 14 patients of seroperitoneum, 10 patients of biliary fistula, 2 patients of massive blood loss during liver resection and 2 patients of re-bleeding after operation.</p><p><b>CONCLUSION</b>The method of hepatic vein occlusion with Satinsky clamp was safe and effective.</p>

Clinical application of hepatic venous occlusion for hepatectomy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Most liver resections require clamping of the hepatic pedicle (Pringle maneuver) to avoid excessive blood loss. But Pringle maneuver can not control backflow bleeding of hepatic vein. Resection of liver tumors involving hepatic veins may cause massive hemorrhage or air embolism from the injuries of the hepatic veins. Although total hepatic vascular exclusion can prevent bleeding of the hepatic veins effectively, it also may result in systemic hemodynamic disturbance because of the inferior vena cava being clamped. Hepatic venous occlusion, a new technique, can control the inflow and outflow of the liver without clamping the vena cava.</p><p><b>METHODS</b>A total of 71 cases of liver tumors underwent resection with occlusion of more than one of the main hepatic veins. All tumors involved the second porta hepatis and at least one main hepatic vein. Ligation or occlusion with serrefines, tourniquets and auricular clamps were used in hepatic venous occlusion.</p><p><b>RESULTS</b>Of the 71 patients, ligation of the hepatic veins was used in 28 cases, occlusion with a tourniquet in 26, and occlusion with a serrefine in 17. Right hepatic veins were occluded in 38 cases, both right and middle hepatic veins in 2, the common trunk of the left and middle hepatic veins in 24, branches of the left and middle hepatic veins in 2, and all three hepatic veins in 5. Thirty-five cases underwent hemihepatic vascular occlusion, 4 alternate hemihepatic vascular occlusion, 23 portal triad clamping plus selective hepatic vein occlusion, and 9 portal triad clamping plus total hepatic vein occlusion. The third porta hepatis was isolated in 26 cases. The amount of intraoperative blood loss averaged (540 +/- 283) (range 100 to 1000) ml in the group of total hemihepatic vascular occlusion and in the group of alternate hemihepatic vascular occlusion, (620 +/- 317) (range 200 - 6000) ml in the group of portal triad clamping plus selective or total hepatic vein occlusion. All tumors were completely removed.</p><p><b>CONCLUSIONS</b>Hepatic venous occlusion applied in hepatectomy can prevent bleeding and air embolism, and is safe and effective with stable hemodynamics.</p>

Comparative study of selective hepatic vascular exclusion and Pringle maneuver in hepatectomy involving the second porta hepatis / 中华外科杂志

<p><b>OBJECTIVE</b>To compare the effects of selective hepatic vascular exclusion (SHVE) and Pringle maneuver in resecting the liver tumors involving the second porta hepatis.</p><p><b>METHODS</b>From January 2000 to October 2005, 2100 liver tumors were resected, among which 235 tumors adhered to or were very close to one or more hepatic veins. Both SHVE and Pringle maneuver were used to control the blood loss during the hepatectomy. They were divided into two groups: SHVE group (125 cases) and Pringle group (110 cases). Data regarding the intra-operative and postoperative courses of the patients were analyzed. SHVE group included total SHVE (clamping of the porta hepatis and all major hepatic veins) in 25 cases and partial SHVE (clamping of the porta hepatic and one or two hepatic veins) in 100 cases. Three methods were used to occlude hepatic veins: be ligated with suture, be encircled and occluded with tourniquets and be clamped with Shatinsky clamps directly.</p><p><b>RESULTS</b>There was no difference between the 2 groups regarding the age, sex, tumor size, cirrhosis and HBsAg positive rate, ischemia time and operating time (P > 0.05). Intra-operative blood loss and transfusion requirements were decreased significantly in the SHVE group. Hepatic veins ruptured with massive blood loss in 14 and air embolism in 3 in Pringle group, but there was no massive blood loss and air embolism in SHVE group. Postoperative bleeding, reoperation, liver function failure and mortality rate were higher in Pringle group (P < 0.05), ICU stay and hospital stay were longer in Pringle group (P < 0.05).</p><p><b>CONCLUSIONS</b>SHVE is much more effective than Pringle maneuver for controlling intraoperative bleeding. It can prevent massive blood loss and air embolism resulting from hepatic veins ruptured and can reduce the postoperative complications rate and mortality rate. Clamping the hepatic veins with Shatinsky clamp is safer and easier than encircled and occluded with tourniquets.</p>

Diagnosis and treatment of hepatic focal nodular hyperplasia: report of 114 cases / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the diagnosis and treatment of the hepatic focal nodular hyperplasia (FNH).</p><p><b>METHODS</b>The clinical data of 114 patients with FNH proved by the pathology were analyzed retrospectively.</p><p><b>RESULTS</b>FNH occurs as a single node in 98.1%, ranging from 0.9 cm to 15.0 cm in diameter [average, (4.2 +/- 2.2) cm], only 2 patients had more than one FNH nodules. 86.0% of patients with FNH were below 50 years old. 89.5% of patients were asymptomatic. AFP was negative in all patients. Hepatitis B was negative in 4.4% (5/114) of patients. Of these patients, 50 lesions were in the left lobe, 50 in the right lobe, 6 in the mid lobe, and 8 in the caudate lobe. A big central artery was found in 3.5% (4/113) of the lesion in patients by color Doppler ultrasound. CT scan showed transient immediate enhancement in 94.3% (66/70) of patients, with central scar in 10 cases. The demonstration of a central scar in the lesion was very helpful for the diagnosis of FNH. MRI demonstrated early vigorous enhancement in 91.8% (56/61) of patients. It is strong enhancement on arterial phase and slight or obvious enhancement on portal venous phase and slightly delayed enhancement of the central scars in FNH. Eleven cases showed central scar. MRI was more helpful for the diagnosis of FNH using liver specific contrast agents: superparamagnetic iron oxide (SPIO). All patients underwent focus resection, and there was no mortality and severe complication.</p><p><b>CONCLUSION</b>FNH shows some typical clinical and imaging features. We could improve the correct diagnosis rate by comprehensively analyzing the clinical and imaging materials.</p>

A case control study on the impact of CYP450 MSPI and GST-M1 polymorphisms on the risk of lung cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the correlation of polymorphisms of CYP1A1 MSPI and glutathiones S-transferase (GST-M1) independently and in combination with the risk of lung cancer.</p><p><b>METHODS</b>A case control study which included 91 cases of lung cancer and 138 controls collected from the First Affiliated Hospital of Sun Yat-sen University of Medical Sciences, Guangzhou Tumor Hospital and The Red Cross Hospital of Guangzhou or conmunity area. All subjects were investigated with a uniform questionnaire. Blood samples were collected from all cases and controls for detecting CYP1A1 MSPI and GST-M1 polymorphisms which were analyzed by PCR and RFLP.</p><p><b>RESULTS</b>It showed that there was no significant difference in frequencies of this genotypes of CYP1A1 MSPI between the two groups. The frequency of GST-M1 null (0/0) genotype was higher in the case group than in the control group, with an OR of 1.38 (95% CI 0.81 - 2.38), but there was no statistical significance. However, combination of several genotypes was strongly associated with lung cancer. There was a synergistic interaction between the m2m2 genotype of CYP1A1 MSPI and GST-M1 (0/0) genotype, with an OR of 2.47 (95% CI 1.03 - 5.90).</p><p><b>CONCLUSION</b>The combination of two genetic polymorphisms significantly increases the risk of lung cancer.</p>

Study on the replication of Hepatitis B Virus in primary hepatocytes from heterogenous species / 中华肝脏病杂志

<p><b>OBJECTIVES</b>By studying the possibility and degree of the replication and expression of human hepatitis B virus (HBV) genes in normal liver cells from heterogenous species, such as primary duck hepatocytes (PDH) and primary rat hepatocytes (PRH), to investigate the species-specificity of HBV infection and replication.</p><p><b>METHODS</b>PDH and PRH isolated by in situ perfusion with low concentration collagenase were transfected with complete HBV genome by electroporation (transfection group, about 1.19 10(12) copies of linear HBV DNA/1 10(7) PRH/PDH). From 1 day to 15 days after transfection, HBsAg and HBeAg in the supernatants and lysates of PRH/PDH were measured with IMX System, HBcAg was assayed with western blotting, Immunol dot blotting and Immunocytochemistry. Moreover, HBV S-mRNA and X-mRNA were tested with RT-PCR. Meanwhile, replicative intermediates of HBV DNA were analyzed by Southern blotting and Dot blotting. PRH/PDH electroporated only was used as control group.</p><p><b>RESULTS</b>HBsAg in the lysates of transfected PDH group were 15.24 (1day), 14.55 (3 days) and 5.13 ( 5 days; P/N values, positive>or=2.1), HBeAg all was negative (<2.1), and both were negative in the supernatants of transfected group. Viral antigen production in transfected rat hepatocytes: HBsAg in the lysates of transfected hepatocytes was positive, P/N values ranging from 2.17 to 93.41, The average P/N values was 14.74+/-31.82, and could be maintained for 15 days after transfection. Whereas, HBsAg in the supernatants of transfected group was only found positive on 1 day after transfection, which P/N value was 6.66. HBeAg and HBcAg in the lysates of PRH of transfection group were positive during the first 3 days following transfection, P/N values was around 2.8. The total amount of HBV DNA in the transfected PDH and PRH groups was strongly positive by dot blotting, whereas that of the control group was negative. Southern blot analysis of intracellular total HBV DNA indicated that there were relaxed circular (RC), covalently closed circular (ccc) and single-stranded (SS) HBV DNA replicative intermediates in the transfected PDH and PRH groups, there was no integrated HBV DNA in the cellular genome. Control groups were negative at all.</p><p><b>CONCLUSION</b>Expression of HBV genes and production can occur in hepatocytes from nonmammalian species or mammalian species, which strongly supports the idea that replication of HBV has no critical species-specificity, and yet it depends on the endoenvironment of hepatocyte.</p>

Study on the angiogenic ability of tumor fibroblasts / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To explore the role of fibroblasts derived from tumor in the tumor angiogenesis.</p><p><b>METHODS</b>Two-well co-culture system were used to detect the expression of MMP-9, TGF-beta1, TN and bcl-2 in L929-H22 cells, and their ability of promoting angiogenesis of ECV304 cells and invasion of MDA-MB-231 cells respectively, which were established in our laboratory before. Then their adhesion and the effect of their supernatant on H22 cells proliferation were analysed.</p><p><b>RESULTS</b>Compared with L929 cells, the adhesion potential of L929-H22 cells increased (F>or=104.32, P<0.001), with the higher level of expression of MMP-9, bcl-2, TN, and TGF-beta1 in L929-H22 cells in creased (t>or=3.3055, P<0.01). L929-H22 and L929 cells enhanced the invasiveness of human mammary cancer MDA-MB-231 cells through artificial basement membrane (Matrigel) 1.21 and 0.48 times respectively (F=266.3, P<0.001). L929-H22 cells induced morphogenesis of ECV304 cells. L929-H22 stimulated endothelial cells to form more and longer tubes than L929 did (F>or=23.75, P<0.01). 25% CM of L929-H22 cells stimulated the growth of H22 cells (F=266.30, P<0.05).</p><p><b>CONCLUSION</b>The results suggested that fibroblasts in tumors secrete more growth factors and angiogenic factors to promote the angiogenesis and invasion of solid tumors.</p>

Long-term health-related quality of life in chronic hepatitis B patients / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate their long-term outcome and the efficacy and economic significance of antiviral drugs by investigating the long-term health-related quality of life (HQL) in chronic hepatitis B (CHB) patients.</p><p><b>METHODS</b>The HQL of 101 CHB patients with biopsy-proven 6 to 18 years ago and 105 persons of general population as control was studied with revised SF-36 questionnaire.</p><p><b>RESULTS</b>The HQL in CHB patients was lower than that in general population in physical functioning, role physical, general health, mental health, and specific symptoms (mu > or = 2.10, P<0.05).</p><p><b>CONCLUSIONS</b>The long-term HQL in chronic hepatitis B patients is poor.</p>

An in vitro model of hepatitis B virus gene replication and expression in primary rat hepatocytes transfected with circular viral DNA / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To establish an in vitro model of hepatitis B virus (HBV) replication and expression in primary rat hepatocytes (PRH) transfected with circular viral DNA for further study on the interaction of HBV with hepatocytes.</p><p><b>METHODS</b>Circular viral DNA containing complete HBV genome were transfected into PRH by electroporation (transfected group, about 4mug of circular viral DNA/1 10(7)cells). From day 1 to day 10 after transfection, HBsAg and HBeAg in the supernatants and lysates of PRH were measured with IMX system. HBcAg was assayed with western blotting, immunol dot blotting and immunocytochemistry. Meanwhile, HBV S-mRNA and X-mRNA were tested with RT-PCR, and replicative intermediates of HBV DNA were analyzed by southern blotting and dot blotting. Moreover, Transmission electron microscopy was used if viral particles were produced in transfected rat hepatocytes. PRH electroporated only was used as control group.</p><p><b>RESULTS</b>(1) Viral antigen production in transfected rat hepatocytes: HBsAg in cell lysates was positive. P/N values ranged from 4.83 to 85.69, and could be maintained for 10 days after transfection. The average P/N values was 18.239 27.459. Whereas, HBsAg was negative in the supernatants of transfected group (P/N values, negative<2.1). HBeAg in the supernatants and lysates of transfected hepatocytes all was negative (P/N values<2.1) during 10 days following transfection. HBcAg was only found positive in transfected hepatocytes by immunol dot blotting. (2) Detection of viral transcripts: transcription of HBV DNA was investigated by preparing total RNA from rat hepatocytes 2 days after transfection and looking for S-mRNA and X-mRNA by RT-PCR. Results showed S-mRNA positive, X-mRNA negative. (3) HBV DNA replication analysis: intracellular total DNA was extracted 2 days after transfection and analysed by southern blotting. All replicative DNA intermediates, including relaxed circular (rcDNA), covalently closed circular (cccDNA), and single-stranded (ssDNA) linear HBV DNA forms, were indicated. Dot blotting showed intracellular HBV DNA positive in transfected group during 10 days after transfection. However, viral particles were not found in transfected hepatocytes during 3 days after transfection.</p><p><b>CONCLUSIONS</b>Circular HBV DNA transfected into primary adult rat hepatocytes could obtain continuous replication and stable expression of HBV surface antigen. This in vitro model has high reproducibility and stability, and is useful for directly studying the interaction of HBV with hepatocytes.</p>

A study on the long-term outcome of hepatitis B e antigen-negative chronic hepatitis B compared with that of hepatitis B e antigen-positive chronic hepatitis B / 中华传染病杂志

0.05)between the two groups.Conclu- sion The long-term outcomes of e-CHB is not markedly different compared with that of e+CHB.