OBJECTIVETo observe the effect of Angong Niuhuang Pill (ANP) on Thl/Th2 in cerebral infarction patients and to explore the mechanism of phlegm-heat obstructing orifices.

METHODSRe- cruited were 30 cerebral infarction patients of phlegm-heat obstructing orifices syndrome (PHOOS) both in China and Indonesia. They were assigned to 4 groups according to the use of ANP, the Chinese treatment group, the Indonesia treatment group, the Chinese control group, and the Indonesia control group. Patients in the two control groups received conventional treatment, while those in the two treatment group additionally took ANP for 30 successive days. Their adverse reactions were observe, and levels of INF-γ and IL-4 were detected.

RESULTSThe INF-γ level and the INF-γ/IL-4 ratio significantly decreased, and the IL-4 level increased after treatment in the four groups with statistical difference (P < 0.05). Compared with the control group after treatment in the same country, the INF-γ level and the INF-γ/IL-4 ratio were lower, and the IL-4 level was higher in the two treatment groups (P < 0.05). Compared with the two Chinese groups, the INF-γ level and the INF-γ/IL-4 ratio were higher, and the IL-4 level was lower in the two Indonesian groups (P < 0.05). There was no statistical difference in the post-treatment indices between the two treatment groups.

CONCLUSIONSANP had moderating effect on Th1/Th2 in cerebral infarction pa- tients. Cerebral infarction patients of PHOOS might exist certain relation with Th1/Th2.

Cerebral Infarction ; drug therapy ; China ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hot Temperature ; Humans ; Indonesia ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism

In this paper 3 different media (A,for yeast cultivation ; B, for laccase producing; D, for white rot fungi cultivation) were compared in enriching and screening decolorizing fungi culture using Reactive Black 5 (RB5) and Reactive Red (M-3BE) from the following three points: decolorization effects, abilities of producing enzymes and diversity of microbial community. 11 groups of fungi with obvious decolorization effects were obtained after enrichment for near one month. Among them, 6 groups came from medium D, the other two 3 groups from medium A and B, respectively. However, the 3 groups from medium A exhibited the highest microbial diversity and best decolorization results with 99.53% and 97.42% color removal rate of Reactive Red M-3BE and Acid Red. From them, 16 strains of fungi were isolated and primarily identified as Saprolegniaceae, Eurotiaceae (Monascus went), Erysiphaceae and Physodermataceae. Fungi groups from medium B and D exhibited a bit lower color removal rate of various dyes and only 3 and 2 isolates primarily classified as Saccharomycetaceae and Eurotiaceae (Penicillium) were obtained from them. Fungi cultures in medium A and B could produce lignin peroxidase, and those in medium D could be detected higher activity of laccase. All the fungal cultures exhibited very weak activity of manganese dependant peroxidase.

OBJECTIVETo establish and evaluate a BA-ELISA method for the quantitative detection of cystic fibrosis transmembrane conductance regulator (CFTR) protein.

METHODSWe deliberately selected three tables of CFTR and made the synthetic peptide be expressed in E. coli, then used the antigen to immunize rabbits to obtain the anti-CFTR polyclonal serum. After that, 96 well plates were coated with the purified antibody against CFTR. The antigen CFTR which was extracted from human sperm was detected by anti-CFTR antibody labeled with biotin, horseradish peroxidase conjugated avidin, and the substrate. The concentrations of two kinds of antibodies and the experiment parameters were optimized. Thereby, the double antibody sandwich BA-ELISA method for the quantitative detection of CFTR protein was established. Furthermore, the reproducibility, specificity and so on were evaluated by clinical specimens of sperm.

RESULTSThe optimal concentration of coated anti-CFTR IgG was 4 µg/ml, while the biotin labeled anti-CFTR IgG was 10 µg/ml; the optimal blocking buffer was 1% BSA-PBST, the optimal time of the reaction between antigen and antibody was 60 min, the optimal chromogenic time was 15 min, the intra-assay and inter-assay coefficient were 2.16%-9.23% and 2.29%-11.71% respectively; The lowest detectable limit was 0.15 ng/ml; the standard curve had a good linear correlation of R2 = 0.962.

CONCLUSIONThe BA-ELISA method for the quantitative detection of CTFR protein is successfully established, and it is demonstrated that the method has strong specificity, high sensitivity and good reproducibility. It provides the basis and evidence of the further application of the method.

Animals ; Antibodies ; Cystic Fibrosis Transmembrane Conductance Regulator ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; Humans ; Peptides ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo investigate the effect of in vivo administration of rhG-CSF and/or rhIL-11 on mice immune system function.

METHODST cell subgroups, suppressor T cells (CD8+ CD28-, CD4+ CD25+, CD3+ CD4- CD8- T cells), expression of CD28 on T cells, and spleen T cells intracellular IL4/IFN-gamma secretion were determined by multicolor flow cytometry. MTT was used to determine the T cell proliferation capacity and mixed lymphocyte reactions.

RESULTSIn vivo administration of cytokines decreased the percentage of lymphocytes (P < 0.05), rhIL-11 and rhG-CSF in combination significantly decreased the CD4+/CD8+ ratio and increased the percentage of CD8+ CD28- suppressor T cells compared to either cytokine alone (P < 0.01). There was no difference in the percentage of CD3+ CD4- CD8- and CD4+ CD25+ suppressor T cells between either of the cytokines. Furthermore, cytokines treatments significantly decreased the capacities of splenic T cells proliferation and the response to alloantigens compared with the PBS treatment (P < 0.05), the combination group being more significantly decreased (P < 0.01). And cytokines treatment significantly decreased the production of IFN-gamma and increased the production of IL-4 compared with the PBS treatment(P < 0.05). The ratio of IFN-gamma/IL-4 were significantly decreased after the combination compared with either of them alone.

CONCLUSIONThe combination of rhIL-11 and rhG-CSF is potentially synergistic in the induction of immune tolerance by their effects on the proliferation capacity and function of T lymphocytes.

Animals ; CD28 Antigens ; metabolism ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Immune Tolerance ; Interleukin-11 ; pharmacology ; Mice ; Mice, Inbred C57BL ; Recombinant Proteins ; T-Lymphocytes ; drug effects ; immunology ; metabolism

Background Human leucocyte antigen (HLA)-B27-associated uveitis is one of the most common causes of non-infectious uveitis.T helper 17 (Th17) cells play an important role in human autoimmune diseases,but the pathology research on the production of Th17 cells in acute anterior uveitis patients positive for HLA-B27 was rarely reported.Objective The aim of this study was to investigate the expression and significance of the peripheral blood T helper cell subsets (Th1,Th2,Th17) in acute anterior uveitis patients positive for HLA-B27.Methods This study meets the criteria of the Helsinki Declaration.Informed consent was obtained from all the participants.A prospective cohort design was used in this study.Twenty-two patients with acute anterior uveitis positive for HLA-B27 were enrolled from Affiliated Second Hospital of Nanjing Medical University,and 16 normal healthy subjects with matched gender and age were enrolled as controls.The expression of interferon-γ (IFN-γ),interleukin-4 (IL-4) and IL-17 on lymphocytes (CD4+) in blood were assessed by flow cytometry,and immunoturbidimetry was used to detect the C reactive protein (CRP) level in blood.The degree of the severity of disease was evaluated by clinical scoring.The correlations between the percentage of IFN-γ+Th1,IL-4+Th2,or IL-17+Th17 with clinical factors and CRP were analyzed.Results The percentages of IFN-γ+Th1 and IL-17+Th17 in the peripheral blood were (23.11 ±9.69) ％ and (3.96±2.92) ％ in the patient group,showing a significant increase in comparison with (16.00±4.26)％ and (1.68±0.60) ％ in the control group (P=0.041,P=0.002).However,the IL-4+Th2 level was not significantly different between the patient group (0.33％ ±0.36％) and the control group (0.56％ ±0.34％) (P=0.122).No significant correlations were found between the percentage of IFN-γ+ Th1 with disease severity and CRP (r =0.197,P =0.500 ; r =0.253,P =0.383),between the percentage of IL-4+ Th2 with disease severity and CRP (r =0.068,P =0.817 ; r =0.439,P =0.116) as well as between the percentage of IL-17 + Th17 with CRP (r =0.226,P =0.436).However,a positive correlation was seen between the percentage of IL-17+ Th17 with disease severity (r =0.805,P =0.001).Conclusions IFN-γ and IL-17 in human CD4+T cells are significantly elevated in the blood of HLA-B27-related acute anterior uveitis patients.Disease severity is associated with the percentage of IL-17 +Th17,suggesting that Th1 cells together with Th17 cells participate in the pathogenesis of the disease and Th17 cells might play a dominant role in the disease.

Myeloid-derived suppressor cells (MDSCs) play a crucial role in T cell dysfunction, which is related to poor outcome in patients with severe trauma. Cyclooxygenase-2 (Cox-2) contributes to immune disorder in trauma and infection via production of prostaglandin E2. However, the role of Cox-2 in the accumulation and function of MDSCs after traumatic stress has not been fully elucidated. In the present study, we treated murine trauma model with NS398, a selective Cox-2 inhibitor. Then the percentages of CD11b+/Gr-1+ cells, proliferation and apoptosis of CD4+ T cells were determined. Arginase activity and arginase-1 (Arg-1) protein expression of splenic CD11b+/Gr-1+ cells, and delayed-type hypersensitivity (DTH) response were analyzed. The results showed that Cox-2 blockade significantly decreased the percentages of CD11b+/Gr-1+ cells in the spleen and bone marrow 48 and 72 h after traumatic stress. NS398 inhibited arginase activity and down-regulated the Arg-1 expression of splenic CD11b+/Gr-1+ cells. Moreover, NS398 could promote proliferation and inhibit apoptosis of CD4+ T cells. It also restored DTH response of traumatic mice. Taken together, our data revealed that Cox-2 might play a pivotal role in the accumulation and function of MDSC after traumatic stress.

Objective To summarize the first 16 eases in mainland China and to discuss the cli-nical experience of robot-assisted laparoseopie radical prostateetomy(RLRP). Methods Sixteen pa-tients with localized prostate carcinoma underwent RLRP with da Vinci S surgical system (Intuitive Surgical Inc.). The age of the patients was 62-76 years, average 69 years. The preoperative t-PSA level was 0.2-79. 2. Ng/ml. The volume of prostate was 9.8-232.9 ml. Fifteen patients were with biopsy-proven prostate cancer, the average Gleason score was 7(4-9). Three were T2a. N0 M0, 4 were T2b N0 M0 and 8 were T2c N0/M0 by clinical stage. One was prostatic intraepithelial neoplasm-Ⅲ. The level of t-PSA in serum and the result of urinary continence were followed up after RLRP. Results All the operations were accomplished successfully. The mean preoperative set-up time of the da Vinci surgical system was 64(60--90)min;the mean operation time was 236(190--390)rain;the mean esti-mated blood loss was 231(50-500)m.L The patients were ambulant between the 2nd and 3rd postop-erative days. Foley catheter was sueeeasfully removed on day 10 to 14, and mean hospital stay was 13 (6-19) days. Two eases had positive surgical margins, the pathological stages were both pT3b N0 M0. The average serum t-PSA was less than 0. 1 ng/ml during a median follow-up of 9(6-12) months. By the conventional definition of urinary continence (0 to 1 pads daily), 94%(15/16) and 100% (16/16)of patients were continent at 3 and 6 months, respectively. Of the patients, 75% (12/16)and 88% (14/16)had no urinary leakage(0 pads daily). Conclusions RLRP is small incision and safe. It is the direction of minimally invasive urologic surgery.

OBJECTIVETo investigate the changes in the disc-condyle-fossa relationship following mandibular advancement.

METHODSTwenty-five patients (10 boys and 15 girls, 11.1 +/- 1.1 years) with class II division 1 malocclusions were treated with Activator. The average treatment time was 10.9 months. Parasagittal MRIs in closed mouth position were analyzed before and after Activator treatment by means of the pair-t test.

RESULTSCa-Tm value [(-0.1 +/- 1.5) mm and (-0.6 +/- 0.9) mm] of the left temporomandibular joint (TMJ) and Ca-Ca' value [(2.3 +/- 0.8) mm and (1.8 +/- 0.5) mm] of the right TMJ were changed significantly after Activator treatment (P < 0.05). This indicated that the condyle was located anteriorly in the fossa. No significant changes were found in the disc-fossa relationship and disc-condyle relationship (P > 0.05).

CONCLUSIONSActivator could promote the growth of the mandible and at the same time, it had no apparent negative influence on TMJ function.

Adolescent ; Child ; Female ; Humans ; Male ; Malocclusion, Angle Class II ; pathology ; therapy ; Mandibular Advancement ; Postoperative Period ; Temporomandibular Joint ; pathology

BACKGROUND:The research and application of platelet-rich plasma in tissue regeneration and restoration have always been an issue of concern in the medicine and bioengineering fields.OBJECTIVE:To analyze the effects of platelet-rich plasma in combination with tendon stem cells on histomorphology change and matrix metalloproteinase 1 expression of the tendon tissues in a rabbit model of Achilles tendinopathy.METHODS:Forty New Zealand white rabbits were randomly divided into model group (n=32) and blank control group (n=8).In the model group,the animals were injected about 2 cm distant to the attachment point of the left calcaneus with prostaglandin E2 (once a week,for totally 4 weeks) to make the animal model of tendinopathy.The rabbits in the blank control group were injected the equal amount of normal saline.After 4 weeks,model rabbits were randomly divided into four subgroups:combination group,tendon stem cell group,platelet-rich plasma group and model control group,with eight rabbits in each group.Platelet-rich plasma and tendon stem cells,alone or in combination,and normal saline were injected into the corresponding group,twice with an interval of 3 weeks.At 6 weeks after injection,the tendon tissue was collected and stained for histological examination and detection of matrix metalloproteinasa 1 expression.RESULTS AND CONCLUSION:(1) Hematoxylin-eosin staining:the tendon fibers in the combinationgroup were intact and arranged orderly;in the tendon stem cell group,the tendon fibers were almost arranged orderly despite some fractured fibers;in the platelet-rich plasma group,fiber breakage and loose fiber structure were observed;in the model control group,there were no intact tendon fibers,with the presence of inflammatory cell filtration.(2) Masson staining:The tendon fibers in the combination group had slight wave-shaped changes but the fibers were not cut off;in the tendon stem cell group,the tendon fibers were slightly in disorder,but with the intact structure,and obvious inflammatory cell filtration was observed;in the platelet-rich plasma group,fiber breakage,reduced collagen fibers and inflammatory cell filtration were obviously observed;in the model control group,there were no intact tendon fibers,and inflammatory cell filtration was clearly visible.(3) The expression of matrix metalloproteinase 1:Compared with the blank control group,the expression of matrix metalloproteinase 1 was significantly higher in the other groups except the combination group (P ＜ 0.05).There was no significant difference in the expression of matrix metalloproteinase 1 between the combination group and blank control group (P ＞ 0.05).To conclude,the combination of platelet-rich plasma and tendon stem cells can inhibit the vicious cycle of degeneration of collagen and extracellular matrix,reduce the expression of matrix metalloproteinase 1 in tenocytes,and delay inflammation responses and degeneration due to tendinopathy.

Objective To observe the long term effects of arthroscopic knee debridement and reconstructing operation for treating osteoarthritis in patients with Kaschin-Beck disease. Methods Thirty-one cases of patients with Kaschin-Beck disease were followed for 6 years after operation of articular clearing by arthroscope. Index of pain, symptoms of self-evaluation, range of motion, walking distance, standing test by affected leg when bending at 30° or 60° were recorded and compared with the preoperative results. Results Twenty-four cases were followed up for 6 years. Six years after operation the pain index(3.38 ± 2.87) was dramatically decreased compared to that before operation (6.88 ± 1.45, t = 5.30, P ＜ 0.05). Patients symptoms markedly improved by subjective self-evaluation was 70.83% (17/24), the effective rate was 100% (24/24). The number of cases that could stand up when leg bending at 30° or 60° were 21,18 cases, respectively, compared with that of preoperative of 14, 11 cases, respectively, the difference was statistically significant(x2 = 5.17,4.27, all P ＜ 0.05). Six years after operation the walking distance(3 cases ＜ 1 km, 11 cases 1 - 5 km and 10 cases ＞ 5 km) were greatly improved compared to the results before operation (12 cases ＜ 1 km, 9 cases 1 - 5 km and 3 cases ＞ 5 km, U = 2.88, P ＜0.05). Six years after operation the knee activity[(132.25 ± 14.52)°] remained at the same level, compared with that of preoperative [(131 .58 ± 14.68) °], the difference was not statistically significant (t = 0.16, P ＞ 0.05) .Conclusions The method of arthroscopic joint debridement to cure Kaschin-Beck disease knee osteoarthritis can significantly reduce pain, improve function and walking distance, with more stable long-term satisfactory outcome.