Asiatic acid inhibits liver fibrosis by blocking TGF-beta/Smad signaling in vivo and in vitro.

Liver fibrosis is a major cause of liver failure, but treatment remains ineffective. In the present study, we investigated the mechanisms and anti-hepatofibrotic activities of asiatic acid (AA) in a rat model of liver fibrosis induced by carbon tetrachloride (CCl(4)) and in vitro in TGF-beta1-stimulated rat hepatic stellate cell line (HSC-T6). Treatment with AA significantly attenuated CCl(4)-induced liver fibrosis and functional impairment in a dosage-dependent manner, including blockade of the activation of HSC as determined by inhibiting de novo alpha smooth muscle actin (a-SMA) and collagen matrix expression, and an increase in ALT and AST (all p<0.01). The hepatoprotective effects of AA on fibrosis were associated with upregulation of hepatic Smad7, an inhibitor of TGF-beta signaling, thereby blocking upregulation of TGF-beta1 and CTGF and the activation of TGF-beta/Smad signaling. The anti-fibrosis activity and mechanisms of AA were further detected in vitro in HSC-T6. Addition of AA significantly induced Smad7 expression by HSC-T6 cells, thereby inhibiting TGF-beta1-induced Smad2/3 activation, myofibroblast transformation, and collagen matrix expression in a dosage-dependent manner. In contrast, knockdown of Smad7 in HSC-T6 cells prevented AA-induced inhibition of HSC-T6 cell activation and fibrosis in response to TGF-beta1, revealing an essential role for Smad7 in AA-induced anti-fibrotic activities during liver fibrosis in vivo and in vitro. In conclusion, AA may be a novel therapeutic agent for liver fibrosis. Induction of Smad7-dependent inhibition of TGF-beta/Smad-mediated fibrogenesis may be a central mechanism by which AA protects liver from injury.

Advanced glycation end-products induce tubular CTGF via TGF-beta-independent Smad3 signaling.

Advanced glycation end-products (AGEs) can induce expression of connective tissue growth factor (CTGF), which seems to promote the development of diabetic nephropathy, but the exact signaling mechanisms that mediate this induction are unknown. Here, AGEs induced CTGF expression in tubular epithelial cells (TECs) that either lacked the TGF-beta1 gene or expressed dominant TGF-beta receptor II, demonstrating independence of TGF-beta. Furthermore, conditional knockout of the gene encoding TGF-beta receptor II from the kidney did not prevent AGE-induced renal expression of CTGF and collagen I. More specific, AGEs induced CTGF expression via the receptor for AGEs-extracellular signal-regulated kinase (RAGE-ERK)/p38 mitogen-activated protein kinase-Smad cross-talk pathway because inhibition of this pathway by several methods (anti-RAGE antibody, specific inhibitors, or dominant negative adenovirus to ERK1/2 and p38) blocked this induction. Overexpressing Smad7 abolished AGE-induced Smad3 phosphorylation and CTGF expression, demonstrating the necessity for activation of Smad signaling in this process. More important, knockdown of either Smad3 or Smad2 demonstrated that Smad3 but not Smad2 is essential for CTGF induction in response to AGEs. In conclusion, AGEs induce tubular CTGF expression via the TGF-beta-independent RAGE-ERK/p38-Smad3 cross-talk pathway. These data suggest that overexpression of Smad7 or targeting Smad3 may have therapeutic potential for diabetic nephropathy.

[Immune and inflammation confusion in severe sepsis and effects of bi-immunomodulation therapy: a prospective, randomized, controlled clinical trial].

OBJECTIVE: To investigate the immune and inflammation confusion state in severe sepsis and the effects of two way immunomodulation therapy with continuous blood purification (CBP), thymosin alpha1, and combined therapy of CBP and thymosin alpha(1). METHODS: 91 Patients with severe sepsis aged > 18, with Marshall score>5. were randomly divided into 4 groups: CBP Group (n = 22) undergoing continuous renal replacement therapy (CRRT) or molecular adsorbents recirculating system (MARS) therapy once a day for 3 days in addition to classical Surviving Sepsis Campaign (SSC) therapy, Thymosin alpha(1) Group (n = 23) undergoing subcutaneous injection of thymosin alpha(1) 1.6 mg once a day for 7 days in addition to SSC therapy, Combined Therapy Group (n = 22) undergoing CBP combined with thymosin alpha(1) treatment in addition to SSC therapy, and SSC Group (treatment control group, n = 24) undergoing SSC therapy only. Peripheral blood samples were collected before treatment, and 3 and 7 days after the beginning of treatment (days 4 and 8) to detect the serum interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-alpha. The levels of CD(14)(+) monocyte human leucocyte antigen (HLA)-DR and T lymphocytes were monitored. The mechanical ventilation time, ICU stay length, and mortality within 28 d and mortality within 90 d were observed. Ten healthy persons were used as healthy control group. RESULTS: Thirty-four of the 91 patients died within 28 d with a mortality of 77.4% (Death Group) and other 57 patients were put in Survival Group. The levels of serum IL-6, IL-10, and TNFalpha, and IL-6/IL-10 at different time points of both Death and Survival Groups were all significantly higher, and the HLA-DR level, and CD(3)(+), CD(4)(+), and CD(8)(+) T lymphocyte numbers at different time points of both Death and Survival Groups were all significantly lower than those of the healthy controls (P < 0.05 or < 0.01). The levels of serum IL-6, IL-6/IL-10, TNFalpha, HLA-DR, and CD(3)(+), CD(4)(+), and CD(8)(+) T lymphocyte at different time points of Death Group were all significantly higher than those of Survival Group (P < 0.05 or < 0.01). The CD(3)(+) T lymphocyte number on day 8 of Thymosin Group was significantly higher than that of SSC Group (all P < 0.05). The serum IL-6 and TNFalpha and IL-6/IL-10 were decreased, and HLA-DR, and CD(3)(+), CD(4)(+), and CD(8)(+) were increased significantly on day 8 in CBP and Combined Therapy Groups. The level of TNFalpha decreased, and the numbers of CD(3)(+) and CD(4)(+) T lymphocytes increased significantly on day 4 in Combined Therapy Group (P < 0.05 or P < 0.01). Compared with Thymosin Group, almost all the indexes of CBP and Combined Therapy Groups were improved, only the CD(3)(+) T lymphocyte level on day 4 increased and the IL-6/IL-10 ratio on day 8 was decreased significantly in Combined Therapy Group (both P < 0.05). Compared with those of SSC Group, the mechanical ventilation time, length of ICU stay within 28 days, and 28 days mortality and 90 days mortality of the 3 treatment groups were all decreased, and there were statistical differences in the length of ICU stay of CBP Group and in the mechanical ventilation time and length of ICU stay within 28 days of Combined Therapy Group (both P < 0.05). CONCLUSION: Systemic inflammatory response and immunodepression exist simultaneously in severe sepsis. Thymosin alpha(1) increases the cellular immunity, and CBP bi-modulates the immune turbulence, reduces the inflammatory mediators, and ameliorates the immune homeostasis. These 2 therapies also improve the clinical prognosis and the combination of both would be more effective.

[Clinical application of xenogenic (porcine) acellular dermal atrix (ADM) in scar treatment].

OBJECTIVE: To investigate the role of xenogenic (porcine) ADM as dermal substitute in scar treatment. METHODS: After scar excision, the wounds were covered with composite grafts of DR procine ADM and autologous thin split-thickness grafts in one stage or in two stages. RESULTS: 22 out of 47 cases were treated in two-staged procedure. After the ADMs were applied to the wound, the autologous thin split-thickness grafts were implanted 7 days later. 25 cases were treated in one-staged procedure. The survival rates of composite grafts were (88.3 +/- 3.7)% for subcutaneous recipient bed and (89.7 +/- 3.4)% for deep fascia recipient bed in group with two-staged procedure, compared with (92.5 +/- 4.1)% and (93.2 +/- 5.2)%, respectively, in group with one-staged procedure. Early after grafts taken, the grafts had a pink colour and smooth surface. The patients were followed up for 90 days at most. The survived composite grafts were durable, elastic, smooth and soft with good function and appearance like normal skin. They could even be pinched up. The scar along the edge of the grafts was slightly hypertrophic. CONCLUSIONS: The survival rate of composite graft is higher in patients with one-staged procedure. The elasticity and textural of the taken grafts are better on subcutaneous recipient bed than on deep fascia recipient bed, though the function has no difference. Xenogenic (porcine) ADM can be an optimal dermal substitute for wound coverage after scar excision.

T3/T4 thoracic sympathictomy and compensatory sweating in treatment of palmar hyperhidrosis.

BACKGROUND: Compensatory sweating (CS) is one of the most common postoperative complications after thoracic sympathectomy, sympathicotomy or endoscopic sympathetic block (ESB) for palmar hyperhidrosis. This study was conducted to examine the relevance between CS and the sympathetic segment being transected in the surgical treatment of palmar hyperhidrosis, and thus to detect the potential mechanism of the occurrence of CS. METHODS: Between October 2004 and June 2006, 163 patients with primary hyperhidrosis were randomly divided into two groups, T(3) sympathicotomy (78 patients) and T(4) sympathicotomy (85), who were operated upon under general anesthesia via single lumen intubation and intercostal video-mediastinoscopy (VM). RESULTS: No morbidity or mortality occurred. Palmar hyperhidrosis was cured in all patients. Follow-up (mean (13.8 +/- 6.2) months) showed no recurrence of palmar hyperhidrosis. The difference of rates of mild CS in groups T(3) and T(4) was of no statistical significance. The rate of moderate CS was significantly lower in group T(4) than in group T(3). No severe CS occurred. CONCLUSION: The rates of occurrence and severity of CS are lowered with the lower sympathetic chain being transected.

A phase II clinical trial with cytotropic heterogeneous molecular lipids (CHML) for patients with hepatic malignancies.

Hepatocellular carcinoma (HCC) and other forms of metastatic liver cancer (MLC) have poor outcomes due to the limited treatment options. Surgery, radiotherapy and chemotherapy have a limited success. Thus, there is an urgent need for novel therapies for patients with advanced HCC and MLC. The response and toxicity profile of a novel biological anticancer agent, cytotropic heterogeneous molecular lipids (CHML), in 135 Asian patients with hepatic malignancies treated at five different hospitals in China from April 1998 to August 2003 is described. This trial included 97 patients with HCC and 38 with MLC. The majority of these patients had received conventional therapies and many had failed to respond or relapsed. CHML was administered by intra-arterial (i.a.) infusion with or without simultaneous intravenous (i.v.) infusion for 25 days with a rest of 2-4 weeks between each cycle. Fifty three percent of patients received two cycles, and 47% received three cycles. The complete response (CR) rates were 23% for HCC and 29% for MLC with an overall CR of 24%. The overall partial response (PR) was 53%. The patients with earlier stages and limited tumor burden had a better response, but a few patients with advanced disease also achieved PR. The patients who achieved CR or PR had a significant increase in long-term survival for up to five years. The treatment with CHML resulted in minimal toxicity and the reported adverse reactions were not higher than grade II. CHML is an effective therapy for hepatic malignancies, showing responses and increases in survival in patients in whom other therapies have failed. CHML is well tolerated and is an excellent candidate for Phase III clinical trials.

[Intercostal video-mediastinoscopy: a report of 701 cases].

OBJECTIVE: To summarize the experience of intercostal video-mediastinoscopy (VMS) in treatment for mediastinal masses, malignant pleural effusion and palmar hyperhidrosis. METHODS: The clinical data of 701 patients received intercostal VMS from November 2001 to June 2007 were summarized retrospectively. Forty-eight patients with mediastinal masses and 46 patients with suspected malignant pleural effusion underwent intercostal VMS pleural biopsy (39 cases with talc pleurodesis) and 607 patients with palmar hyperhidrosis underwent bilateral intercostals VMS thoracic sympathectomy. RESULTS: No mortality and morbidity were reported in this group. Definitive pathologic diagnosis had been made through VMS mediastinal masses biopsy in mediastinal masses and pleural biopsy in pleura effusion. The efficiency of talc pleurodesis was 100% for 39 cases. The symptoms of sweating of hands in 607 patients with palmar hyperhidrosis disappeared completely, all patients' hands became dry with a 1.5 degrees C to 3.0 degrees C increase of the skin temperature immediately after operation. No recurrence occurred during the follow-up. CONCLUSION: VMS is a simple, convenient and alternative procedure for the treatment of mediastinal masses, malignant pleural effusion and palmar hyperhidrosis.

[Treatment of deep partial thickness burns by a single dressing of porcine acellular dermal matrix].

OBJECTIVE: To explore the effect of one dressing of porcine acellular dermal matrix on deep partial thickness burns. METHODS: From January 1997 to January 2004, sixty-seven cases of deep partial thickness total burned surface area (TBSA) from 50% to 90% burn wound were treated by a single dressing of porcine acellular dermal matrix (the porcine acellular dermal matrix group). Ten cases of deep partial thickness burned patients with the same TBSA treated by exposure method served as the exposure method group. The healing time of the wound was observed. The patients were followed up for 3 months to 2 years, and the scar proliferation was observed. RESULTS: The deep partial-thickness wound would be healed without dressing change in the porcine acellular dermal matrix group, and the average healing time was (12.2 +/- 2.6) days. The average healing time of the exposure method group was (27.4 +/- 3.5) days. Follow up of the patients within 3 months to 2 years showed that scar proliferation in the porcine acellular dermal matrix group was much less than that in the exposure method group, even no scar proliferation was observed in some patients. CONCLUSION: Without tangential excision, autografting and dressing change, a single dressing of porcine acellular dermal matrix on deep partial thickness burn wound could shorten the healing time and inhibit scar proliferation.

[Observation on hybrid bioartificial liver support systems in treating chronic severe hepatitis: a study of 60 cases].

OBJECTIVE: To study the clinical efficacy of three kinds of hybrid bioartificial liver support systems (HBLSS) in treating chronic severe hepatitis. METHODS: A bioartificial liver support system (BAL), comprising porcine hepatocytes and fiber tube style bioreactor, was constructed. Then three kinds of HBLSS were constructed: Molecular absorbent recirculating system (MARS) plus BAL; slow plasma exchange (SPE) plus continuous hemodiafiltration (CHDF) and BAL; and SPE plus hemoperfusion (HP) and BAL. One hundred-twenty patients in middle or late stages of chronic severe hepatitis were enrolled in this study. They were randomly divided into 6 groups: H1 group was treated with BAL+MARS, H2 with BAL+SPE+CHDF and H3 with BAL+SPE+HP (as treatment groups); C1 group was treated with MARS, C2 with SPE+CHDF and C3 with SPE+HP (as control groups). The changes in the clinical symptoms, in the hepatic encephalopathy stages, and in the serum total bilirubin (TBIL), the serum albumin (ALB), the prothrombin activities (PTA), endotoxin, ammonia, creatinine and a-fetal protein (AFP) were all observed before the treatment, right after it and 72 hours later. The improving and curing rates and the rates of side effect occurrences in each group were observed. RESULTS: In all 6 groups, the patients' clinical symptoms ameliorated; their TBIL, endotoxin and ammonia levels decreased (P<0.05), and their PTA and AFP levels lowered significantly (P<0.05). But in the H1, H2 and H3 groups they were more distinctive than in the control groups. In H1 and H2 groups creatinine and ammonia levels were decreased more significantly than in the H3 group (P<0.05). The improving and curing rates of each group were 65 % (13/20), 60% (12/20), 45% (9/20), 45% (9/20), 40% (8/20) and 20% (4/20) respectively. No serious side effects were observed during the treatment. CONCLUSION: In treating middle and late stage chronic severe hepatitis, the measures used in H1, H2 and H3 are better than those in C1, C2 and C3. Furthermore, H1 and H2 treatments can ameliorate hepatic and renal functions, prevent the development of multiple organ dysfunction syndrome, and are better than those used in H3.

[Some medical staff positive for serum SARS coronavirus antibody IgG have only mild symptoms].

OBJECTIVE: To identify patients with SARS coronavirus infection who have only mild symptoms. METHOD: Enzyme-linked immunosorbent assay was employed to detect serum antibody against SARS coronavirus in the lysate of whole SARS coronavirus from 19 SARS patients and 200 medical staff members without obvious SARS symptoms after possible exposure to the virus during routine medical practice. RESULTS: Serum IgG antibody against SARS coronavirus was detected in all the 19 SARS patients, and among the 200 staff members, 20 (10%) were found positive for the antibody but with no obvious or only mild symptoms. CONCLUSION: Serum IgG antibody against SARS coronavirus is positive in a small proportion (around 10%) of the medical staff members exposed to the virus in our hospital, but may not cause obvious symptoms, suggesting SARS coronavirus infection might in some cases have mild or even no clinical manifestations.

[Development and application of a new hepatitis C virus genotyping method with polymerase chain reaction-reverse blot dot technique].

OBJECTIVE: Using polymerase chain reaction-reverse blot dot (PCR-RDB) technique to establish a new method for hepatitis C virus (HCV) genotyping and to study the distribution of HCV genotypes in Foshan area. METHODS: HCV primers and probes were designed in 5'-untranslated region (nt-1-nt-299) of HCV. HCV RNA in serum was isolated and purified, and its cDNA was obtained by reversed transcription. Nested PCR using biotin-labelled primers, was done. PCR products were hybridized with immobilized specific probes (genotype 1a to 3b) on Biodyne C membrane to genotype HCV by color development while adding POD and TMB. A certain judgment could be made according to the position of color reaction. The reliability of this new method was verified by sequencing. HCV RNA levels in serum were determined by real time fluorescent quantitative (FQ)-PCR. 60 FQ-PCR-positive HCV sera from Foshan area were genotyped using this assay. RESULTS: All 60 sera could be successfully genotyped by PCR-RBD. 50 (83.3%) cases were found to be genotype 1b, 2 (3.3%) as genotype 1a and 2 (3.3%) as genotype 2a while 5 (8.0%) to be mixture of genotype 1a and 1b, and 1 (1.7%) to be mixture of genotypes 1b and 2a. No genotypes 2b, 3a and 3b were found. The results of PCR-RDB genotyping methods coincided with sequence analysis. CONCLUSION: Newly established HCV genotyping system was proved to be sensitive, specific, precise and economic, thus suitable for clinical and epidemiologic studies. The results of HCV genotyping showed that genotype 1b was the predominant genotype in Foshan area.

[Establishment of a new HBV genotyping method with PCR-RBD and its application].

OBJECTIVE: Using PCR-RDB to establish a new method for HBV genotyping, and to survey the distribution of HBV genotypes in the Foshan area. METHODS: Biotin-labeled primers for amplification of HBV region X (nt1550-1789) were used to amplify extracted HBV DNA. HBV was genotyped by hybridization of the PCR products with immobilized specific probes (genotype A to F) on C membrane. Color development was achieved by adding POD and TMB. A judgment was made according to color reactions. The reliability of this new method was verified by gene sequencing. 300 samples of HBV DNA-positive sera from the Foshan area were genotyped using this assay. RESULTS: Of the 300 sera genotyped by PCR-RBD, 147 (49.0%) cases were genotype B, 136 (45.3%) were genotype C, 1 (0.3%) genotype D, and 12 (4.0%) were mixtures of genotype B and C, and 4 (1.3%) were mixtures of genotype C and D. No genotype A, E or F were found. The results of PCR-RDB genotyping were consistent with the results obtained with sequence analysis. CONCLUSION: This newly established HBV genotyping system proved to be sensitive, specific, precise and economic, and should be suitable for clinical practice and epidemic study. The results of HBV genotyping show that genotype B and C are the predominant genotypes in the Foshan area.

[The first case of severe acute respiratory syndrome in Foshan].

OBJECTIVE: To evaluate the epidemiologic feature, diagnosis and treatment of severe acute respiratory syndrome (SARS). METHODS: To describe the epidemiologic and clinical features of the first case of SARS in Foshan city, Guangdong province retrospectively, and to review the diagnostic procedure. RESULTS: This case had the following features: (1) a history of contact with mild cats and eating the animal's meat; (2) high fever (temperature, > 38 degrees C), followed by dry cough, rapid progression to respiratory failure, followed by radiographic evidence of bilateral air-space lesions; (3) no leukocytosis; (4) spread to 4 family members who had had direct contact with this patient; (5) the patient's serum SARS virus IgG was confirmed to be positive; (6) the patient was treated with anti-viral agents, antibiotics and mechanical ventilation and molecular adsorbent re-circulating system (MARS). CONCLUSIONS: The clinical manifestations of the SARS case, which was highly infectious, met world health organization (WHO) criteria for the diagnosis of SARS. Early initiation of mechanical ventilation and supportive therapy contributed to the good prognosis of this critical case.

[Farm compost polluted water may induce pharyngo-esophageal, gastric and liver carcinoma in the chicken].

OBJECTIVE: To study farm compost polluted water that may induce pharyngo-esophageal, gastric and liver carcinoma in chickens. METHODS: 280 chickens were randomized into 4 groups: experiment group 100 chickens fed with compost water + NaNO(2) by stomach tube. The other 180 were evenly randomized into 3 control groups (60 each), fed with compost water, NaNO(2) and tap water in the same way. The farm compost was prepared with corn stalks, rice straws, excreta of men and livestock. The compost water, after being nitrosified and acidified, was fed through stomach tube 5 - 7.5 ml/session, twice a week. Besides, a solution consisting of the respective formula of each group added with 3 - 4 L water with pH adjusted to 3 - 4 by 1N HCL was given ad lib to all chickens in each group for 26.5 months. RESULTS: In the experiment group, there were pharyngo-esophageal carcinoma 16 (16.3%), gastric adenocarcinoma 5 (10.4%) and liver carcinoma 3 (6.3%), in contrast to none in the 3 control groups, showing significant differences (P < 0.01, P < 0.01, P < 0.05). CONCLUSION: Successful simulation of the layout of esophageal carcinoma high morbidity area and the mimic of chicken gastric fluid strongly support our compost etiological hypothesis that the nitrosified and acidified compost water are carcinogenic, very well causing esophageal, gastric and liver carcinoma.

[Porcine acellular dermal matrix in the treatment of deep partial-thickness burns in human].

OBJECTIVE: To investigate the possibility of treating deep partial-thickness burns by closed dressing of the wounds with porcine acellular dermal matrix (ADM) and evaluate the therapeutic effects. METHODS: We conducted a retrospective review of 128 cases of burn patients who received treatment with porcine ADM within the period from January 1998 to January 2002 in our hospital. Different procedures were adopted according to the degree of the burn injury. As for "fairly superficial" deep partial-thickness skin burns, after removing the necrotic epidermis and washing with 0.1 % benzalkonium bromide, the wound was covered with porcine ADM pretreated with povidone-iodine and then bandaged with cotton gauze and bandages. In cases of "fairly deep" deep partial skin thickness burns, eschar excisions as deep as to expose parabiotic lamina were performed prior to dressing the wounds in the same manner as described above. RESULTS: All the patients were successfully treated with satisfactory clinical results. CONCLUSIONS: Porcine ADM is feasible as an efficient dressing material for deep partial-thickness burns, which may promote epithelialization in the wounds and help stabilize the patients' condition during burn shock stage to reduce the complications and shorten the treatment courses.