Human tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) is a member of the tumor necrosis factor (TNF) family of ligands which has been reported in 1995. The TRAIL protein induces apoptosis of certain types of target cells, such as transformed cells that include but are not limited to cancer cells and virus-infected cells but the normal cells. It is a type II transmembrane protein and the extracellular domain of TRAIL is the functional domain in induction of cell apoptosis. A gene fragment encoding for the active domain of TRAIL was modified with oligo-nucleotide directed mutagenesis according to the characters of Pichia pastoris expressing vector. Arginine at the position of 149 corresponding to the amino acid residue 531 which might be a potential Kex2 protease processing sites was substituted with Lysine to prevent the expressed protein from the digestion by the protease. After proved with DNA sequencing. the modified gene fragment coding soluble TRAIL domain was inserted into the Pichia pastoris expression vector pPIC9K in the same reading frame with alpha-factor secreting signal peptide. The recombinant plasmid pPIC9K - TRAIL was transferred into P. pastoris cell by spheroplast transformation. The recombinant yeasts were identified by antibiotic G418 and Southern dot blot. The transformants (His+ Mut(s)) containing multi-copy gene fragment of TRAIL were selected with increasing concentration of G418 and induced with 0.5% methanol in shaking flask to expression the active domain of TRAIL. After inducing for 3 - 4 days, the proteins in the culture supernatant was assayed with SDS-PAGE and Western blot. Two expressed protein bands whose appearant molecular weight were 19kD and 38kD, respectively, could be specifically recognized by polyclonal antibodies against human TRAIL. The 38kD protein might be a dimers of TRAIL in the culture supernatant. The amount of expressed foreign protein made up to 36% of the total proteins in the culture suprenatant. Biological activity assay, in vitro, indicated that the expressed protein could induce tumor cells apoptosis.
Apoptosis ; drug effects ; Cell Line, Tumor ; Electrophoresis, Polyacrylamide Gel ; Genetic Vectors ; genetics ; Humans ; Immunohistochemistry ; Pichia ; genetics ; metabolism ; Protein Structure, Tertiary ; genetics ; physiology ; TNF-Related Apoptosis-Inducing Ligand ; genetics ; metabolism ; pharmacology

OBJECTIVETo explore the correlation between pathological characteristics of target organs and excess evil syndrome in IgA nephropathy.

METHODSData were collected in multicenter cooperation. Totally 266 IgA nephropathy patients were typed into exogenous wind-heat affection syndrome (49 cases), lower energizer damp-heat syndrome (100 cases), damp-phlegm syndrome (43 cases), and blood stasis syndrome (74 cases). Meanwhile, percutaneous renal biopsy was performed in all patients for Hass classification, Oxford classification, Katafuchi integral, and Jiang's classification methods. The correlation between excess evil syndrome and pathological index was analyzed.

RESULTSFour syndrome types were correlated with their Hass levels (r = 0. 341, P <0. 01). Affection of exogenous wind-heat syndrome was correlated with segmental proliferation of endothelial cells and damaged active lesions of segmental capillary loops. Lower-energizer damp-heat syndrome was associated with Hass III level, destroying active lesions of capillary loops, segmental proliferation of endothelial cells, glomerular segmental lesions, focal interstitial infiltration of inflammatory cells, focal interstitial fibrosis and tubular atrophy. Blood stasis syndrome was associated with Hass IV level, glomerular sclerosis, segmental glomerulosclerosis (S)/adhesion, mesangial hypercellularity (M), angiohyalinosis, multi-foci interstitial infiltration of inflammatory cells, multi-foci interstitial fibrosis and tubular atrophy. Phlegm-damp syndrome had higher proportions of Hass I and III levels, but with no association with other pathological parameters.

CONCLUSIONSExcess evil syndrome was associated with partial pathological characteristics of IgA nephropathy. It could reflect pathological damage degree of target organs, activities, chronic lesions, and prognosis of IgA nephropathy to certain extent. Correlated pathological characteristics and its evolution could indicate excess evil syndrome types and their evolution rules.

Capillaries ; Fibrosis ; Glomerulonephritis, IGA ; pathology ; Glomerulosclerosis, Focal Segmental ; Humans ; Kidney Glomerulus ; Medicine, Chinese Traditional ; Prognosis ; Syndrome

OBJECTIVETo investigate the natural history and clinical outcomes in a cohort of transfusion-related hepatitis C in northern areas of China. Methods The authors studied 283 patients (137 males, 146 females, mean age 45.79+/-9.92 age) who became infected with HCV while donating plasma 12.25 years ago. These cases were subjected to ultrasonography and liver biochemical tests and serologic anti-HCV assays. Statistical analyses were performed using the SPSS software.

RESULTSUltrasonographic findings suggestive of liver cirrhosis were 8.3% of cases. No decompensated cirrhosis or HCC was detected. The value of ALT was higher in severe chronic hepatitis (mean 62.07+/-50.87 IU/L) and cirrhosis (mean 115.50+/-108.41 IU/L) patients than in the other groups (mean 32.30+/-29.10 IU/L). The abnormal rate of ALT was 53.3% in severe group, 100% in cirrhosis group.

CONCLUSIONThe natural history of transfusion-related hepatitis C in the areas seemed to be relatively mild when compared with previous data. Our cases showed relatively low rate of positive findings in ultrasonography. None of this cohort had the decompensated cirrhosis or HCC. The factor of sex but not the age at time of infection was found being related to the outcomes. In the absence of liver biopsy, ultrasonography was a suitable and sensitive method for the diagnosis of the progressive hepatitis and cirrhosis.

Adult ; Aged ; China ; epidemiology ; Female ; Hepatitis C, Chronic ; diagnostic imaging ; epidemiology ; transmission ; Humans ; Liver ; diagnostic imaging ; Liver Cirrhosis ; complications ; diagnostic imaging ; Male ; Middle Aged ; Transfusion Reaction ; Ultrasonography

This paper introduces the old nine-needle including its name, shape, function, manipulation and origination, as well as systematically introduces the new nine-needle as a new product which has been developed under the guidance of modern science and technology based on the old nine-needle since 1949. As a new tool, the new nine-needle provide much more choice for acupuncture treatment and obtained satisfied clinical effect. During the development of research and clinical practice on the new nine-needle, its theory has been consummated successively. Combining various kinds of needles for treatment is not only paying attention to the specificity of one kind of needles, but also emphasizing the whole function of all kinds of needles, which makes the new nine-needle formed the characteristic and distinct theory of its own.
Acupuncture Therapy ; history ; instrumentation ; methods ; China ; History, Ancient ; Needles

With the unique structure and manipulation techniques of blood-letting and cutting, hook needle serves as a role in dredging meridians, removing blood stasis, purging heat for resuscitation and relaxing synechial tissues. There are three needling techniques of hook needle: swift pricking, bleeding and pricking. Six manipulations are commonly used as lifting-thrusting, plucking, pulling, pushing-scraping, vibration and massage. The mechanism and manipulations of hook needling are summarized systematically in the present paper. In this way, its manipulations, indications and contraindications are expounded in detail.
Acupuncture Therapy ; instrumentation ; methods ; Humans ; Needles

OBJECTIVE: To examine the genetic polymorphism of 9 STR loci in 5 ethnic groups (including Tu, Sala, Dongxiang, Baoan and Yugu) in Gansu and Qinghai, and to evaluate its application. METHODS: Nine STR loci (D3S1358, FGA, TH01, D7S820, VWA, CSF1PO, D5S818, D13S317 and TPOX) were selected as genetic markers. With STR compound amplification and genescan methods, in which STR loci were marked by fluorescence, the genotype of 5 ethnic groups were examined in 606 unrelated individuals by ABI 377 sequencer. These parameters, such as polymorphism information content (PIC), heterozygosity (H), discrimination power (DP) and probability of paternity exclusion (PPE) were calculated. RESULTS: The genotype frequencies of the 9 STR loci were in accordance with Hardy-Weinberg equilibrium. PIC was within 0.6054 - 0.8735, H was within 0.6158 - 0.8736, DP was within 0.7964 - 0.9691, and PPE was within 0.4610 - 0.8838. Cluster analysis based on allele frequencies in genesis showed Tu, Sala, Dongxiang and Baoan ethnic groups were very close, but Yugu was a little bit far. There were obvious gene exchanges among the populations in north and south of China. CONCLUSION All the 9 STR loci are highly polymorphic in the 5 ethnic groups, which can be useful genetic markers in forensic medicine and population genetics.
China ; ethnology ; Cluster Analysis ; Ethnic Groups ; genetics ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Polymorphism, Genetic ; Tandem Repeat Sequences ; genetics

Objective: To investigate the effects of artesunate treatment on chronic graft-versus-host disease (cGVHD). Methods: Recipient BALB/c mice received 8 × 10⁶ bone marrow cells with 8×10⁶ spleen cells from B10D2 mice. Artesunate solubilized in acetone was injected intraperitoneally every day at the dose of 1 mg/kg at Day 28 after BMT. The clinical scores, survival and histopathological damage were analyzed. The frequency of Th17 and Tregs in PB and spleens from the mice were evaluated by flow cytometry. In addition, CD4⁺ T cells from the spleens of mice were cultured in vitro, then stimulated with artesunate, the frequency of Th17 and Tregs in these splenocytes were evaluated by flow cytometry. Results: Artesunate administration diminished clinical and histopathological damage, and improved the survival of cGVHD mice[(46.57±7.83)% vs (55.71±6.99)%, χ²=5.457, P=0.020]; Artesunate contributed to Tregs development [(4.45±0.04)% vs (8.40±0.23)%, t=15.679, P<0.001; (6.62±0.24)% vs (10.48±0.48)%, t=6.587, P=0.003] while decreased Th17 cells [(1.51±0.18)% vs (0.58±0.19)%, t=7.233, P<0.001; (1.48±0.38)% vs (0.71±0.18)%, t=3.653, P=0.011] expressions in both PB and spleens, and decreased the Th17/Treg ratio (0.34±0.05 vs 0.09±0.03, t=7.621, P=0.002; 0.19±0.03 vs 0.06±0.02, t=6.993, P=0.002). Moreover, artesunate suppressed the Th17 cells expressions [(0.82±0.37) % vs (3.39±1.22) %, t=4.044, P=0.007] and contributed to Tregs development [(34.63±1.29) % vs (14.28±1.69) %, t=19.119, P<0.001], and also decreased the Th17/Treg ratio (0.24±0.09 vs 0.02±0.01, t=4.780, P=0.003) in vitro. Conclusions Artesunate suppressed the Th17 cells expressions and contributed to Tregs development, which provided new sights into the development of a novel drug for cGVHD, e.g., artemisinin.

A comparison of recognition to migraine and treatment methods for it was made between traditional Chinese medicine and western medicine in this paper. Emphasis was made on standardized manipulation processes and techniques of sharp hooked needling for treating the disease, based on both the organic conception of the human body, the theory held by TCM, and on the principle of treatment according to pathogenesis obtained through differentiation of symptoms and signs. The necessary preparation before operations, methods for its application and announcements in the standardized manipulations of sharp hooked needling were all explained in detail. Based on its needling, pricking, dissecting and relaxing effects, the mechanisms of sharp hooked needling in treating the disease were investigated. It is concluded that sharp hooked needling is very effective in treating migraine.
Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; standards ; Humans ; Medicine, Chinese Traditional ; instrumentation ; methods ; standards ; Migraine Disorders ; therapy ; Needles ; Reference Standards

Objective To explore a training system of high level professionals with international vision and meeting the develop-ment of assistive technology in China. Methods The training system was constructed in the theory framework of International Classification of Functioning, Disability and Health,and work process-oriented. Results and Conclusion A curriculum system with tree-type modes and a practice system with three-level and multi-form of assistive technology specialty were constructed,that was recognized by industry and society in the developing.

Objective: To investigate the effects of artesunate treatment on chronic graft-versus-host disease (cGVHD). Methods: Recipient BALB/c mice received 8 × 10(6) bone marrow cells with 8×10(6) spleen cells from B10D2 mice. Artesunate solubilized in acetone was injected intraperitoneally every day at the dose of 1 mg/kg at Day 28 after BMT. The clinical scores, survival and histopathological damage were analyzed. The frequency of Th17 and Tregs in PB and spleens from the mice were evaluated by flow cytometry. In addition, CD4(+) T cells from the spleens of mice were cultured in vitro, then stimulated with artesunate, the frequency of Th17 and Tregs in these splenocytes were evaluated by flow cytometry. Results: Artesunate administration diminished clinical and histopathological damage, and improved the survival of cGVHD mice[(46.57±7.83)% vs (55.71±6.99)%, χ(2)=5.457, P=0.020]; Artesunate contributed to Tregs development [(4.45±0.04)% vs (8.40±0.23)%, t=15.679, P<0.001; (6.62±0.24)% vs (10.48±0.48)%, t=6.587, P=0.003] while decreased Th17 cells [(1.51±0.18)% vs (0.58±0.19)%, t=7.233, P<0.001; (1.48±0.38)% vs (0.71±0.18)%, t=3.653, P=0.011] expressions in both PB and spleens, and decreased the Th17/Treg ratio (0.34±0.05 vs 0.09±0.03, t=7.621, P=0.002; 0.19±0.03 vs 0.06±0.02, t=6.993, P=0.002). Moreover, artesunate suppressed the Th17 cells expressions [(0.82±0.37) % vs (3.39±1.22) %, t=4.044, P=0.007] and contributed to Tregs development [(34.63±1.29) % vs (14.28±1.69) %, t=19.119, P<0.001], and also decreased the Th17/Treg ratio (0.24±0.09 vs 0.02±0.01, t=4.780, P=0.003) in vitro. Conclusions: Artesunate suppressed the Th17 cells expressions and contributed to Tregs development, which provided new sights into the development of a novel drug for cGVHD, e.g., artemisinin.
Animals ; Artesunate ; Graft vs Host Disease ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes, Regulatory ; Th17 Cells