Preventive effect of Ganfujian granule on experimental hepatocarcinoma in rats

AIM:To investigate the inhibitory effect of dietary and medicinal formula Ganfujian granule on diethylnitrosamine (DEN)-induced hepatocarcinoma in rats.METHODS: Male SD rats had free access to water containing 0.1g/L DEN for 16 weeks, during which the rats fed with standard diet or administration of Ganfujian granule (30.4g/Kg in diet). At weeks 4,8,12 and 16 of hepatocarcinogenesis 5 rats of each group were sacrificed, and at week 20 another 30 rats were sacrificed from each group. The end point for survival observation was at week 28.Immunochemistry methods were used to examine the effect of Ganfujian granule on the process of hepatocarcinogenesis including proliferation of hepatocytes and cell cycle modulation.RESULTS:Ganfujian granule could reduce and delay the incidence of hepatocarcinoma in rats and prolong the survival of animals. In addition, Ganfujian granule had a marked inhibitory effect on high expression of cyclin dependent kinase (CDK4) during the whole process of hepatocarcinogenesis and cyclin D1 at week 16 and the number of proliferating cell nuclear antigen (PCNA) positive cells in different stages of hepatocarcinogenesis.CONCLUSION: Ganfujian granule can reduce and delay the incidence of hepatocarcinoma in rats by exerting direct or indirect effects on cell cycle and inhibiting uncontrolled proliferation of hepatocytes.

Effect of traditional Chinese medicinal enemas on ulcerative colitis of rats

AIM: To investigate the effects of traditional Chine semedicinal enema (TCME) on inflammatory and immune response of colonic mucosa of rats with ulcerative colitis(UC), and to observe the pathogenic mechanism.METHODS: Thirty UC rats, induced by intestinal enema together with 2.4-dinitrochlorobenzene (DNCB) and acetic acid, were randomly divided into 3 groups, i.e., GⅠ, GⅡ and GⅢ. Groups GⅠ and GⅡ were administered with TCME and salazosulfapyridine enema (SASPE), respectively. Group GⅢ was clystered with only normal saline (NSE), served as control. Group GIV was taken from normal rats as reference,once daily, from the 7th day after the establishment of UC for total 28 d. Interleukin-6 (IL-6) in the colonic mucosa was assayed by ^3H-TdR incorporation assay. Colonic mucosal lymphocyte subpopulation adhesive molecules, CD4^+CD11a^+,CD4^+CD18^+, CDs^+CD11a^+, CDs^+CD18^+ (LSAM), tumor necrosis factor (TNF)-α, and interferon-γ (IFN-γ), were detected by enzyme linked immunosorbent assay (ELISA). Moreover, the expression of TNF-α mRNA and IFN-γ mRNA in colonicmuc osa were detected by polymerase chain reaction (RT-PCR).RESULTS: Before therapies, in model groups, GⅠ, GⅡ and GⅢ,levels of IL-6, TNF-α, IFN-γ, CDs^+CD11a^+ and CD8^+CD18^+ were significantly different (38.29+2.61 U/mL, 16.54+1.23 ng/L,8.61+0.89 ng/L, 13.51+2.31% and 12.22+1.13% ,respectively) compared to those in GIV group (31.56+2.47 U/mL, 12.81+1.38 ng/L, 5.28+0.56 ng/L, 16.68+1.41% and 16.79+1.11%, respectively). After therapeutic enemas,in GI group, the contents of IL-6 (32.48±2.53 U/m), TNF-α(13.42±1.57 ng/L) and IFN-γ (5.87+0.84 ng/L) were reduced; then, the contents of CD8^+CD11a^+ (16.01+1.05 %)and CD8^+CD18^+ (16.28±0.19%) were raised. There was no significant difference between groups GⅠ and GIV, but the difference between groups GⅠ and GⅡ was quite obvious (P<0.05). The expressions of TNF-α mRNA and IFN-γ mRNAin group GⅢ were much higher than those of group GIV,but those in group GI were significantly suppressed by TCME therapy.CONCLUSION: Ulcerative colitis is related to colonic regional mucosal inflammatory factors and immune imbalance. TCME can effectively inhibit regional mucosal inflammatory factors and improve their disorder of immunity.

Kangxian ruangan keli inhibits hepatic stellate cell proliferation mediated by PDGF

AIM: To investigate the effect of Kangxian ruangan keli (KXR) on hepatic stellate cell (HSC) proliferation mediated by platelet-derived growth factor (PDGF) and the underlying mechanism.METHODS: In a serum-free culture system, HSCs were treated with a KXR preparation for 24 hours, followed by stimulation with PDGF-BB for 24 hours. Then the cells were incubated again in the medium containing KXR for 3 hours stimulated with PDGF-BB for 5 minutes, and collected. The proliferation of HSC was examined using an MTT assay and flow cytometry. Tyrosine phosphorylation was detected with Western blotting and visualized by the enhenced chemiluminescent (ECL) method.RESULTS: The OD values for the HSCs growing in the media without and with addition of PDGF were 0.17±0.06 and0.82±0.05, respectively. The PDGF-induced increase was hindered remarkably by KXR preparation in a dose-dependent manner. The reaction values for the systems with 5mg/mL, 2.5 mg/mL and 1.25 mg/mL of KXR were 0.28±0.03,0.37±0.02 and 0.43±0.04, respectively. Moreover, the percentages of S-phase cells in these KXR-containing culturesystems were 10.95±1.35, 32.76±1.07 and 43.19±1.09,respectively, all of which were significantly lower than that in the culture free of KXR (68.24±2.72). In addition, the values for tyrosine-phosphorylated protein in HSCs treated with 5 mg/mL and 1.25 mg/mL of KXR were 0.1349±0.0072 and 0.1658±0.0025, respectively, which were smaller than that in the cells treated only with PDGF-BB (0.1813±0.0117).CONCLUSION: Within the dose range used in the present study, KXR preparation shows an inhibitory effect on HSC proliferation induced by PDGF. The mechanism of this process may involve interference with tyrosine phosphorylation mediated by PDGF.

Traditional Chinese medicine syndromes of chronic hepatitis B with precore mutant

ABM: This study aims at exploring the distribution of TCM syndromes in CHB patients with HBV pre-core mutation (1896) and the relationship between pre-core mutation and T lymphocytes subgroup, through which to provide objective data on clinical syndrome differentiation of TCM, and further to suggest the therapeutic principle and guide clinical treatment. METHODS: One hundred and forty CHB patients were evenly divided into two study groups, HBV pre-core mutant group and HBV pre-core wild-type group. Besides, 30 healthy blood donors were selected as a healthy control group. HBV-labeled compound, T lymphocytes subgroup, and HBV-DNA pre-core mutant were tested in the study groups. T lymphocytes subgroup were also tested in the control group. All the patients were both diagnosed by syndrome differentiation of TCM and western medicine. RESULTS: The most common syndrome in mutant group was damp-heat combined with blood stasis, and the most common syndrome in the wild-type group was damp-heat stasis in the middle-jiao. There were more cases of medium and severe hepatitis in mutant group than that in wild-type group. The content of CD4+ lymphocytes and CD4+/CD8+ ratio were decreased gradually (healthy control group>wild-type group>mutant group). In the wild-type group, severe and medium CHB patients had considerably lower level of them than mild CHB patients. However, in the mutant group, the opposite result appeared. Meanwhile, the content of HBV-DNA in mutant group was higher than that in wild-type group. CONCLUSION: Damp, heat, toxin and blood stasis were the basic pathogens of CHB, whether pre-core mutant or not. CHB with precore mutant may lead to more severe hepatitis. The decreased content of CD4+ lymphocytes and ratio of CD4+/CD8+ may be taken as one of the indices in confirming the deficiency syndrome of CHB patients with pre-core mutation.

Effect of Weikangning on gastric cancer cell growth and expression of vascular endothelial growth factor and its receptors KDR and Flt-1

AIM: To observe the effect of Chinese traditional herbal decoction Weikang-ning (WKN) on cell growth and expression of VEGF and its receptors KDR and Flt-1 in gastric cancer cell line MGC-803. METHODS: A total of 120 male Wistar rats were divided into control group, high dose, medium dose and low dose groups fed with natural saline, 20, 10, and 5 g/kg of WKN, respectively. The experimental animals were finally killed for the preparation of drug-containing serum. The gastric cancer cell MGC-803 was cultured with the drug-containing serum drawn from the rats in different groups. We observed the growth condition of the cancer cells with light microscope and flow cytometer. The expression of mRNA of VEGF and its receptors KDR and Flt-1 was detected with RT-PCR. RESULTS: The proportion of cells in G0-G1 phase was (65.40±0.41)%, (56.92±0.62)%, (55.89±0.69)% in high, medium and low dose groups respectively vs(41.35±0.55)% in control group (P<0.01), while the cells in G2-S and S phases were (11.62±0.62)% and (22.99±0.69)%, (17.08±0.80)% and (26.00±0.71)%, (19.37±0.57)% and (24.74±0.64)% in high, medium and low dose groups, respectively, vs(23.65±0.56)% and (35.00±0.60)% in control group (P<0.01). The expression of mRNA of VEGF and its receptors was significantly decreased, the area of electrophoresis bands (AREA), the absorptivity of mean optical density (A) and the product of AREA and A were significantly lower in WKN-administered groups than that in control group(P<0.01). CONCLUSION: The decoction of WKN suppresses the growth of gastric cancer cell MGC-803 and decreases the expression of mRNA of both VEGF and its receptors KDR and Flt-1.

Effect of Hejie decoction on T cell immune state of chronic hepatitis B patients

AIM: To explore the effect of Hejie decoction (HJD) (mediation decoction) on T cellular immune state of chronic hepatitis B patients.METHODS: Sixty-five patients with chronic hepatitis B were randomly divided into 2 groups. Forty patients in the treatment group were treated by HJD, and 25 patients in the control group were treated by routine Western medicine. The TCRVβ7 gene expression, T lymphocyte subsets (CD3^+, CD4^+, CD8^+,CD4^+/CD8^+) levels were observed before and after treatment.RESULTS: The level of CD4^+ cells was lower whereas the level of CD8^+ cells was higher in patients than in the normal group. There was no significant difference between the levels of CD3^+ cells in patients and normal persons. After 6 months of treatment, ALT, AST, TB levels of the 2 groups were obviously decreased, and the level of CD4^+ cells was increased whereas the level of CD8^+ cells was decreased in the treatment group. However, the level of CD4^+ cells and CD8^+ cells had no significant difference in the control group. TCRVβ7 expressions were detected in 6 patients of the treatment group, whose HBV-DNA and HBeAg turned negative and ALT became normal. HBeAg in another 3 patients turned negative while HBV-DNA did not, and TCRVβ7 expressions were not detectable. TCRVβ7 expression could not be detected in the control group, HBV-DNA of the control group did not turn negative. HBeAg in 1 patient turned negative while HBV-DNA did not, and TCRVβ7 expressions were not detectable. The total effective rate was not significantly different between the 2 groups and the markedly effective rate was significantly different(P<0.01).CONCLUSION: H3D is effective for treating chronic hepatitis B, and its effect seems to relate with the improvement of the TCRVβ7 expression of chronic hepatitis B patients, thus activating T cells and eliminating HBV. T cellular immune function plays an important role in HBV infection and virus elimination.

Biphenyl Ferulate from Glehnia littoralis

From the underground parts of Glehnia littoralis Fr. Schmidt ex Miquel (Umbelliferae), a new compound named glehnilate was obtained. Its structure was determined by analysis of its spectral data.

Detection of anti-HAV antibody with dot immunogold filtration assay

AIM: To establish a rapid, sensitive and specific immunogold assay for detection of hepatitis A virus infection.METHODS: Rabbit monoclonal antibodies to anti-human IgM and IgG (Dako) were dotted on a nitrocellulose membrane (NCM) respectively to capture the human sera IgM and IgG. Then the captured antibodies would conjugate to HAV antigen, which was revealed by mouse anti-HAV IgG conjugated to gold particles. Final results were assessed by blind method.RESULTS: Sera from 96 patients with acute hepatitis were used for our study. Compared with well-recognized standard (Abbott Laboratory, USA), the sensitivity and specificity of IgM-DIGFA (self-made) were 91.3 % (42/46) and 96.0 %(48/50), and those of IgM-ELISA (Kehua, Shanghai) were 97.8 % (45/46) and 100.0 % (50/50). The identical results were produced from the study with reagents at different conditions, and the study was repeated in 15 negative sera and 10 positive sera. The serum anti-HAV IgG was tested with DIGFA at the same time. In comparison with ELISA,the sensitivity and specificity of DIGFA for IgG anti-HAV were 87.2 % (41/47) and 91.8 % (45/49), respectively.CONCLUSION: This assay can detect anti-HAV IgM and IgG simultaneously, and be done within 3 minutes. The simplicity, rapidity and specificity of the assay were useful for screening and epidemiological study.

Effects of RSM and astragus on expression of Fos and Jun proteins in rat brains after cerebral ischemia and reperfusion

AIM: To investigate the expression of Fos and Jun proteins in rat brains after focal cerebral ischemia followed by reperfusion and effects of RSM and astragus.METHODS: 30 SD adult male rats were divided into 5 groups at random .Group A:sham operated group;Group B:model group;Group C:treated with RSM;Group D:treated with astragus;Group E:treated with RSM and astragus.The immunohistochemistry and medical image processing system(MIPS)were used to measure the numbers and mean grey levels of Fos and Jun protein positive cells in rat cerebral cortex of 5 groups. RESULTS: (1) In cerebral cortex of group B ,C ,D ,E,the numbers of Fos and Jun positive cells were more than those in group A and mean grey levels of Fos and Jun positive cells were lower than those in group A(P< 0.01);(2) In cerebral cortex of ischemic sides in group C,D,E,the numbers of Fos and Jun positive cells were less than those in group B and mean grey levels of Fos and Jun positive cells were higher than those in group B(P< 0.01) ;(3) Group E had more significant effects than group C or group D (P< 0.01). CONCLUSION: The expression of Fos protein and Jun protein in model group increased significantly,compared with sham operated group;RSM ,astragus ,RSM and astragus all could inhibit partly the expression of Fos protein and Jun protein after cerebral ischemia and reperfusion;Prescription of RSM and astragus had stronger inhibiting effects than RSM or astragus.It may be one of mechanisms that ischemic stoke is treated by reinforcing Qi and activating blood circulation therapy in TCM clinic.

New Constituents from the Fresh Stems of Opuntia Dillenii

Aim To study the chemical composition of Opuntia dillenii Haw. Methods Manykinds of chromatogra-phy methods were used in the isolation procedure, while the structures ofisolated compounds were determined on the aids of NMR and MS spectral analysis. Result Three newcompounds, together with 14 known compounds, were isolated form the 80% ethanolic extract of itsstems. Conclusion The three new compounds, opuntioside Ⅰ (2), 4-ethoxyl-6-hydroxymethyl-α-pyrone(3) and kaempferol 7-O-β-D-glucopyranosyl (1→4)-β-D-glucopyranoside (4), were characterized.

Chemical Constituents of Stellera chamaejasme L

A diterpenoid, neostellerin, was isolated from the roots of Stellera chamaejasme L together with4 known compounds identified as β-sitosterol, scopoletin, umbelliferone and daucosterol. The structure ofneostellerin, which was elucidated by NMR, especially 2D NMR analysis, was different from other daphnane typediterpenes isolated from this plant.

Effect of Fuzheng Yiliu decoction combined with chemotherapy on patients with intermediate and late stage gastrointestinal cancer

AIM: To investigate the therapeutic effects of Fuzheng Yiliu (strengthening the body resistance to inhibit tumor) decoction combined with chemotherapy on the patients with intermediate and late stage gastrointestinal cancer. METHODS: Sixty patients were randomly divided into treatment group (chemotherapy combined with Fuzheng Yiliu decoction) and control group (chemotherapy alone). Four indexes, including the tumor recent remission rate (RR), the change of main symptoms, the toxic and side effects caused by chemotherapy and the change of performance status, were observed in the patients. Peripheral blood contents of CD3+, CD4+, CD8+ cells, CD4+/CD8+ and soluble interleukin-2 receptor (sIL-2R) were tested before and after treatment and the values were compared with those of healthy peoples. RESULTS: The improving rate of main symptoms (69.6%) and performance status (56.7%) were significantly higher in the treatment group than in the control group (34.8%, 26.7%, P<0.05). The occurrence rates of grade II toxic and side-effects on both bone marrow (13.3%) and digestive tract (30%) were lower in the treatment group compared to the control group (36.7%, 63.3%, P<0.05). Before treatment, the proportion of CD3+, CD4+ and CD4+/CD8+ decreased and the proportion of CD8+ and SIL-2R raised markedly both in the control group and treatment group as compared to the healttiy people. After treatment, that increased of CD3+, CD4+, CD4+/CD8+ increased (62.25±10.01% vs 68.31±9.72%, 36.83±10.44% vs 42.6±9.62%, 1.24±0.65 vs l.66±0.85, P<0.05) and the values of CD8+ and sIL-2R decreased obviously (33.06±7.69% vs 29.24±6.25%, 588.23±216.86 U/mL vs 475.87±211.36 U/mL,P<0.05) in the treatment group, whereas these values were opposite in the control group (64.22±6.91% vs60.63±5.75%,35.62±7.49% vs31.53±5.53%, 32.95±8.28% vs 37.14±7.48%, 1.17±0.43 vs 0.94±0.43, 573.63±214.32 U/mL vs 692.17?21.33 U/mL, P<0.05). CONCLUSION: Fuzheng Yiliu decoction can enhance therapeutic effects of chemotherapy on malignant gastrointestinal tumor, and also reduce the toxic and side effects on bone marrow and digestive tract, thereby improving the quality of life and cellular immunity in patients with malignant gastrointestinal tumor.

A New Coumestan from Arachis hypogaea L.

A new coumestan, 3, 9-dihydroxy-4, 8-dimethoxycoumestan, was isolated from Arachis hypogaea L. together with two known compounds: 3, 9-dihydroxy-4-methoxycoumestan and 3, 9-dihydroxy-8-methoxycoumestan. The structure was established by spectroscopic methods.

Induction of apoptosis in human T leukemic cell line CEM-6T by tripter-ine;comparison with arsenic oxide and hydroxyl camptothecin

In China, the herbal plant Lei gong teng (tryipterygium wolfordii Hook F) is widely used in rheumatic diseases, indicating that some compounds in this plant might have effects on immune cells. We have reported that one of the compound from this plant, tripterine could induce apoptosis in the human mast leukemic cell line HMC-1 (Chin J of Hematol 20:146,1999). To further explore the potential value of triperterine in the treatment of leukemia, we cultured human T leukemia cell line CEM-6T in vitro with tripterine, or arsenic oxide, or hydroxyl camptothecin at various concentration ranging from 0.0625 μM to 128 μM DNA electrophoresis and annexin V labeling were used to detect apoptosis. The protein levels of Bcl-2,

Traditional Chinese herbal formulate Jin Gui Shen Qi Wan affects the pro-tein levels of Bcl-2 family members and suppresses apoptosis in human lymphocytes

The Bcl-2 knockout mice demonstrated fulminant lymphoid apoptosis, polycystic kidneys, hypopig-mented hair, as well as abnormalities in development, bone formation and life span (Cell 75, 229). These symptoms are typically known as deficiency of kidneys in traditional Chinese medicine. We hypothesize that the Chinese concept of the Kidneys is related to the in vivo roles of the Bcl-2 family and the traditional Chinese herbal formulates designed for affecting the functions of the Kidneys could modulate the expression of this gene family. To test this hypothesis, the blood were collected from the volunteers before and after their taking the Jin Gui Shen Qi Wan (Jin Gui) Pills for Tonifying Kidney-energy, a classic herbal

Experience in the Clinical Application of Zhongzhu(TE 3)

Zhongzhu (TE 3), located on the dorsum of the hand between the 4th and 5th metacarpal bones, in the depression proximal to the metacarpophalangeal joint,1 cun posterior to Yemen (TE 2), is a point on the Sanjiao Channel of Hand Shaoyang. According to the classical documents, this point is mainly indicated for diseases on the head, eye, ear, shoulder, elbow and hand. In the clinical acupuncture practice, however,the author has found that strong stimulation at Zhongzhu (TE 3) can deal with certain acute diseasesor the acute attacks of some chronic diseases. Some of the sample cases are cited in the following.

TRADITIONAL CHINESE MEDICINE IN THE TREATMENT OF AUTOIMMUNE AND IMMUNE DISORDERS

It is well known that the treatment of Dampness-heat falling into the lowerburner(DHFLB) is very difficult to deal with by both traditional Chinese medicine(TCM) and western medicine.After TCM system of Epidemic Febril Disease(composed of three parts,Shanghan,exogenous febril disease;)

REGULATORY ACTION OF ASTRAGALUS SAPONINS AND BUZHONG YIQI COMPOUND ON SYNTHESIS OF NICOTINIC ACETYLCHOLINE RECEPTOR ANTIBODY IN VITRO FOR MYASTHENIA GRAVIS

With overall analysis of symptoms and signs of myasthenia gravis(MG)basing on “Pi(Spleen) Deficiency”(脾虚)theory and with years of our clinical experience in treating MG,we performed a pharmacological study of Astragalus saponins and Buzhong Yiqi Compound(Tonic granulae invigorating vital energy) in 14 peripheral blood mononuclear cell(PBMNC)cultures from 10 MG patients.