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This review summarized the research achievements of mushroom sauce in recent years, from the aspects of raw and auxiliary materials selection, formula, and process research, analysis and testing, existing problems, and prospected the raw material quality and nutrition safety detection.

Researched on micro-hardness of W9Mo3Cr4V high speed steel (HSS) cutting tool material with the impact of pulsed magnetic field. The HSS samples were magnetized with directional pulse on a self-build platform for the magnetic treatment test. The effects of different pulse magnetic field strength, frequency and magnetization time on the micro-hardness of HSS material were studied, and the changes of HSS material microstructure under the action of pulse magnetic field were analyzed by observation metallurgrical structure. Test results indicate that the appropriate pulse magnetization parameter can improve the specimen micro-hardness of HSS material obviously. Pulsed magnetic can make part of the retained austenite translate into hard and brittle martensite inside the metallic material, which makes martensite content increase, and material organization become more uniform. [ABSTRACT FROM AUTHOR]

Objective To construct the lentiviral RNA interference ( RNAi) vectors of rat PER2 gene and evaluate the effects of silencing PER2 gene expression by siRNA. Methods The target sequence of siRNA- PER2 in rats was designed, and the complementary DNA containing both sense and antisense oligonucleotides was synthesized. After phosphorylation and annealing, the double-stranded DNA was cloned to pGLV-U6-EGFP to construct pLentivirus-per2-rat. Then the effects of RNAi in reducing gene expression were further confirmed by real-time PCR in transfected rat A7r5 aortic vessel smooth muscle cells expressing PER2. Results The rat lentivirus vectors expressing PER2-specific shRNA were synthesized, and the specificity of the designed target sequence was primarily identified by electrophoresis and gene sequence analysis. Rat A7r5 aortic vessel smooth muscle cells were successfully transfected with pLentivirus-per2-rat, and the transfection rate was 70%. The real-time PCR analysis confirmed that the level of PER2 mRNA was decreased significantly as compared with that in the negative control group, and suppressed by approximately 84% in cells transfected with pLentivirus-per2-rat. Conclusion The successful construction of rat lentivirus vectors expressing PER2-specific shRNA may lay a foundation for further study on the clock gene's function. [ABSTRACT FROM AUTHOR]

Based on the continuous observation on located trees and branches, the phenology and sexual reproductive process of Xanthoceras sorbifolia Bunge in an annual growth cycle in Wudan area were investigated, and the relationships between fruit growth process of X. sorbifolia and effective accumulative temperature as well as the quantitative dynamics of flowers and fruits were studied. The results showed that the phenology of X. sorbifolia had a significant linear correlation with effective accumulative temperature, and there was a time sequence in the male flower silk elongating and antheral unfolding. The fruits of X. sorbifolia experienced three times of dropping during their growth process. The pollination rate was 26.4%, flower dropping rate was 73.6%, and fruit dropping rate was 92.9%, suggesting that the dropping of flowers and fruits was very serious. The fruits had a quicker vertical growth from early June to mid June and a quicker horizontal growth in the last ten-day of June, with the highest ratio of vertical to horizontal length being 2.17. The individuals of X. sorbifolia had definite differences in their phenology, which was disadvantageous to the cross pollination among the individuals, and the enhancement of pollination rate. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of high mobility group protein 1(HMGB1) on the proliferation and cytokine expression of human bone marrow mesenchymal stem cells (MSC)., Methods: Different concentrations of recombinant human HMGB1 protein (100, 200, 400, 800 and 1000 ng/ml) were incubated with MSC for 24, 48, 72 h and the proliferation of MSC were detected respectively by using the CCK-8 method and flow cytometry. The best concentrations of HMGB1 incubated with MSC was determined (200 ng/ml, 1000 ng/ml), and the flow cytomerty was used to determine the effect of HMGB1 on the proliferation of MSC. The mRNA expression levels of IL-10, TGF- β1, TSG-6 and IFN-γ in MSC incubated with HMGB1 protein were detected by real-time quantitative PCR and ELISA., Results: The result of MSC identification and flow cytometry showed that the CD105, CD73 and CD90 were expressed, but did not expression CD45, CD34, CD11b, CD19 and HLA-DR; CCK-8 showed that HMGB1 at the concentrations of 100 ng/ml, 200 ng/ml and 400 ng/ml could promote the proliferation of MSC incubated for 24, 48 and 72 h as compared with the control group (P<0.05), and the most effective concentration was 200 ng/ml; flow cytometry showed that the compared with the control group, HMGB1 200 ng/ml could induce MSC from G1 phase to S phase to promote the proliferation of MSC; QPCR showed that the mRNA expression of MSC cytokines IL-10, TGF-β1, TSG-6 increased while IFN-γ decreased at the concentration of 200 ng/ml HMGB1 as compared with the control group. ELISA experiments showed that the HMGB1 200 ng/ml acting on MSC for 48 h could significantly promoted the secretion of IL-10, TGF-β 1 and TSG-6(P<0．05), while IFN-γ showed no significant difference as compared with control group., Conclusion: Recombinant human HMGB1 can promote the proliferation and secretion of MSC in healthy people.

Objective: To investigate the expression and clinical significance of high mobility group box 1(HMGB1) in spleen of adult patients with chronic and refractory immune thrombocytopenia(ITP)., Methods: Twenty chronic and refactory ITP patients received splenectomy were enrolled in ITP group and 20 cases of traumatic spleen rupture were enrolled in control group. The splenectomy efficacy in ITP patients was analyzed retrospectively. The HMGB1 expression in spleen tissue was detected by immunohistochemistry, and the correlation between different expression levels of HMGB1 and splenectomy efficacy were analysed. Meanwhile, the protein expression levels of HMGB1 in peripheral blood serum and mononuclear cells(PBMNC) of 25 patients with chronic and refractory ITP were detected by ELISA and Western blot., Results: The median platelet count before splenectomy was 7.5 (0-20) ×10 9 /L; all the patients showed that the initial response to splenectomy within the first month after operation was 100%, the median time of response was 1 day (1-6 days). The median peak platelet count post splenectomy was 448.5 (161-1272)×10 9 /L. In the median time of 10(3-30) months, the platelets count in 8 patients was reduced to varying degrees. After a median follow-up of 69.5 months (22-195), complete response was found in 12 patients, 4 cases showed response and 4 did not. The HMGB1 expression positive rate in spleen of ITP patients was significantly higher than that in control group (85.0% vs 15.0%)(P<0.001). There were a negative correlation between the HMGB1 expression in ITP and therapeutic outcome after splenectomy (r=-0.791, P<0.01). In addition, HMGB1 expression levels in serum and PBMNC of the patients with chronic refractory ITP were also significantly higher than that in healthy controls (P<0.01)., Conclusion: The splenectomy has been found to be effective therapeutic method for patients with ITP, the HMGB1 highly express in the spleen of the patients with chronic refractory ITP, but negatively correlats with the therapeutic outcome after splenectomy.

Objective: To investigate the effects of shRNA targeting mPGES-1 on tumorigenicity of human acute leukemia K562 cells in nude mice in vivo and its mechanisms., Methods: For experiment 3 groups including KD group(expression of mPGES-1 in K562 cells was down-regulated by shRNA), CON (cells without any treatment) and NC group (cells treated with nonspecific-sequence shRNA) were set-up. Western blot was used to test the expression of β-catenin and cyclinD1 in cells. Then the cells of 3 groups were implanted into BALB/c nude mice subcutaneously to establish murine xenograft model. The growth state of the mice and the size of the xenograft tumor were recorded. HE staining was used to observe the morphology of xenograft tumor. Expressions of β-catenin and cyclinD1 in xenograft tumor were detected by immunohistochemical staining., Results: In vitro the expression of β-catenin and cyclinD1 in KD group were lower than the CON group and NC group (P<0.05). In vivo the tumor volume and weight of KD group were significant smaller than the other two groups (P<0.01). HE staining showed that tissues in the KD group were relatively looser in arrangement with smaller cell nucleus and less cytoplasm. The expression of β-catenin and cyclinD1 in the KD group were remarkable weak as compared with that in CON group and NC group (P<0.05)., Conclusion: Down-regulating the expression of mPGES-1 by shRNA may significantly inhibit the tumorigenicity of K562 cells in nude mice in vivo and its mechanism may be related with the inhibition of expression of β-catenin and cyclinD1.

Objective: To explore the role of Treg cells in the pathogenesis of idiopathic thrombocytopenic purpura (ITP)., Methods: The ITP mouse model was established, the Treg cell ratio in peripheral blood and spleen was detected by flow cytometry, the CD4+ CD25+ T cells were sorted by immunomagnetic beads, the Treg cell associated transcription factors (Foxp3, Smad7, STAT5 and Akt-1) and cytokines (IL-10, TGF-β) in CD4+ CD25+ T cells were enriched from spleen mononuclear cells, and the mRNA expression of Treg cell was measured by real-time PCR., Results: The ratio of Tregs in peripheral blood and spleen decreased significantly in ITP mouse, as compared with the controls (P<0.01). In addition, the mRNA expression of IL-10, TGF-β and Foxp3 decreased significantly in spleen CD4+ CD25+ T cells (P<0.05). Expression of Smad 7 mRNA was higher than that of controls., Conclusion: The alteration in Treg frequency and function may be responsible for the immune dysfunction in ITP disease. It is also speculated that the lower mRNA expression of Foxp3 and higher mRNA expression of Smad 7 may inhibit the proliferation and differentiation of Treg cells.

The quantitative identification and enrichment of viable regulatory T cells (Treg) requires reliable surface markers that are selectively expressed on Treg. Foxp3 is the accepted marker of natural Treg, but it cannot be used to isolate cells for functional studies. CD127 is a new surface marker expressed in Treg cells. In this study, two populations of Treg, including CD4(+)CD25(+)CD127(low/-) and CD4(+)CD25(+)Foxp3(+)T cells, and profiles of the Foxp3 expression in CD4(+)CD25(+)CD127(low/-) cells were compared to evaluate which population is better. The peripheral blood cells were collected and spleen suspension of BALB/C mice were prepared, and using triple staining CD4, CD25, CD127 and CD4, CD25, Foxp3. The profiles of Treg, including CD4(+)CD25(+)CD127(low/-) and CD4(+)CD25(+)Foxp3(+) were detected by flow cytometry. The quadruple staining CD4, CD25, Foxp3 and CD127 were used to determine the CD127 expression in CD4(+)CD25(+)Foxp3(+) cells. The results showed that on T cell subset the median expression levels of CD4(+), CD4(+)CD25(+) were 39.02%, 5.35% in peripheral blood and 23.49%, 3.86% in spleen. On CD4(+) T cell subset, the median expression level of CD4(+)CD25(+)CD127(low/-) and CD4(+)CD25(+)Foxp3(+)T cells were 7.13%, 3.97% in peripheral blood and 12.8%, 8.23% in spleen. The ratio of CD4(+)CD25(+)CD127(low/-) T cells was higher than that of CD4(+)CD25(+)Foxp3(+) cells in both peripheral blood and spleen cells (P < 0.01). The CD4(+)CD25(+)CD127(low/-) cells highly expressed Foxp3, while the CD4(+)CD25(+)Foxp3(+)T cells lowly expressed CD127. It is concluded that compared with the CD4(+)CD25(+)Foxp3(+) populations, CD4(+)CD25(+)CD127(low/-) T cells better fit the definition of naturally occurring regulatory T cells in peripheral blood cells and spleen of BALB/C mice. CD127(low/-) is a characteristic marker on surface of CD4(+)CD25(+) Treg cells, and has been confirmed to be more specific marker for quantitatively sorting Treg cells.

To investigate the effect of a microsomal prostaglandin E synthase-1 (mPGES-1) inhibitor MK886 on cell cycle of the human acute myeloid leukemia HL-60 cells. HL-60 cells were treated with different concentration of MK886 (10, 25, 50 µmol/L) for 24 h. Flow cytometry, Western blot and ELISA were used to measure cell cycle, cyclin D1, mPGES-1, PGE(2), Akt, P-Akt and C-MYC. The results indicated that after treated with MK886, the percentage of HL-60 cells decreased in G(0)/G(1) phase and increased in S phase, and expressions of mPGES-1, cyclin D1, P-Akt and C-MYC and synthesis of PGE(2) decreased significantly. It is concluded that MK886 can arrest HL-60 cells in G(0)/G(1) phase, the mechanism of which is possibly associated to inhibition of mPGES-1 expression, reduction of PGE(2) synthesis, suppression of Akt phosphorylation and C-MYC expression, down-regulation of cyclin D1 expression.

This study was aimed to investigate the effect of MK886, a mPGES-1 inhibitor, on apoptosis and drug resistance of leukemia HL-60/A cell line. Expression of mPGES-1 was assayed by QT-PCR and Western blot. The effect of MK886 on HL-60/A cell proliferation was assayed by CCK-8 method, and flow cytometry was used to detect cell apoptosis. The expression of Akt and P-Akt was detected by Western blot. PGE2 was measured by ELISA. Effect of MK886 (10 µmol/L) on the chemotherapeutic sensitivity of HL-60/A cells and expression of mdr-1 mRNA and P170 protein were investigated too. The results indicated the expression of mPGES-1 was higher in HL-60/A cells. MK886 inhibited HL-60/A cell proliferation and induced apoptosis in a time- and concentration-dependent manner. Expression of mPGES-1 and P-Akt and synthesis of PGE2 decreased significantly. MK886 reduced expression of mdr-1 and P170 protein and enhanced the sensitivity of HL-60/A cells to chemotherapeutic drugs. It is concluded that MK886 can inhibit HL-60/A cell proliferation, induce apoptosis and enhance sensitivity to chemotherapeutic drugs, the mechanism of which possibly associates to down-regulation of mPGES-1/PGE2 synthesis, reduction P-Akt expression and decreasing mdr-1 and P170 protein expression.

This study was purposed to investigate the inhibitory effect of bactericidal permeability-increasing protein (BPI) on lipopolysaccharide (LPS)-mediated activation of platelets. Venous blood samples were obtained from 10 healthy volunteers and were prepared into platelet-rich plasma (PRP, 1 × 10(8)/ml). Experiments were divided into four groups: normal platelet group (untreated group); LPS group, BPI group and BPI+LPS group. PRP were stimulated by LPS (10 µg/ml) in the presence and absence of BPI (100 µg/ml) or BPI alone. Then platelets were harvested and determined for Toll-like receptor-4 (TLR-4) with flow cytometry (FCM), the supernatant was used for detection of cytokines including tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) with enzyme-linked immunosorbent assay (ELISA). The results showed that as compared with normal platelet group, TLR-4 expression on platelets was significantly increased under LPS stimulation (P < 0.001); the levels of TNF-α and IL-6 in the supernatant were also remarkably elevated (P < 0.001). However, either TLR-4 expression or the cytokine levels significantly decreased in the presence of BPI when platelets underwent LPS-challenge (P < 0.05), but still were higher than that in normal platelet group. Stimulating the platelets with BPI alone could not enhance the TLR-4 expression and cytokine levels. It is concluded that BPI has the ability to inhibit the LPS-induced platelet activation.

Objective: The aim of this study was to investigate whether miR-449a, miR-449b and miR-192 family microRNAs play the same roles in p53 pathway as miR-34 family in ovarian cancer., Methods: Wild-type p53 ovarian carcinoma cell line A2780 cells were treated with genotoxic agent adriamycin. The reactivation of p53 was detected by Western blot. The expression of miR-449a/b, miR-34a, miR-34b, miR-34c, miR-192 and miR-194 were detected by real-time quantitative PCR. Mutant p53 ovarian cancer cell line SKOV3.ipl cells were transfected with pre-microRNAs and the cell-cycle changes were detected. The expression level of miR-449a/b, miR-34a, miR-34b, miR-34c, miR-192 and miR-194 in serous ovarian carcinomas of varying grade and stage were compared with real-time PCR., Results: The expressions of miR-449a/b, miR-34b and miR-34c were 19-fold to 21-fold elevated after p53 activation by genotoxic agent. Ectopic expression of miR-449b, as well as miR-34c, resulted in cell-cycle arrest in SKOV3.ipl cells. The expression of miR-449a/b was parallel with that of miR-34b, miR-34c, and were significantly lower in late stage and high-grade serous carcinomas than in the normal fallopian tube, early stage and low-grade serous carcinomas. The expression of miR-192, miR-194 and miR-34a did not show evident features in serous ovarian carcinomas and were much lower than miR-449a/b, miR-34b and miR-34c in normal fallopian tube., Conclusions: As tumor-suppressor microRNAs, miR-449a/b, miR-34b and miR-34c cooperate and play important roles in p53 pathway. Their inactivation may contribute to the carcinogenesis and progression of serous ovarian carcinomas.

To observe the expression of cyclin D1, hTERT, and telomerase activity in MNC, HL-60, HL-60A and to explore their effects on leukemogenesis and drug-resistance, normal human peripheral blood mononuclear cells, HL-60 cells sensitive to adriamycin and HL-60A cells resistant to adriamycin were investigated. The cell cycle was analyzed by flow cytometry, and the apoptosis was analyzed by Annexin V-FITC(+) PI staining. Expressions of cyclin D1 and hTERT were determined by real-time PCR and Western blot. Telomerase activity was detected by TRAP-ELISA. The results indicated that the percentage of MNC, HL-60 and HL-60A in S phase was (10.21 + 2.11)%, (44.93 + 3.00)%, and (51.38 + 1.10)% respectively; the percentage of apoptosis cells was (16.14 + 2.13)%, (7.53 + 0.92)%, (4.15 + 0.96)% respectively; the expression of mRNA and protein for cyclin D1 and hTERT increased; the telomerase activities of HL-60 and HL-60A were higher (p = 0.000), whereas the difference between HL-60 and HL-60A was no statistically significant (p = 0.232); positive correlation between cyclin D1, hTERT and telomerase activity had been found (p < 0.01). It is concluded that the cells of S phase increased while the apoptotic cells decreased in HL-60 and HL-60A, especially in HL-60A, which may be due to the up-regulation of cyclin D1, hTERT and telomerase activity.

Objective: To investigate the effects of miR-449 and miR-34 on cell growth, cell cycle and target gene expression based on these miRNA different expressions in ovarian cancer cell lines SKOV3 and SKOV3-ipl both with mutation of p53., Methods: The expressions of miR-449a/b and miR-34b,c in SKOV3 and SKOV3-ipl were detected by RT-PCR. miR-449a,b and miR-34b,c were ectopically expressed by transfection of SKOV3-ipl. The cell growth rate was assayed by MTS method. The changes of cell cycle were measured by FCM. The changes of expression of cell cycle related proteins were detected by Western blot., Results: Ectopic expression of miR-449b and miR-34c resulted in lowered adhesion activities by 28%-34%, and in cell cycle arrests with increased cell number of 15.62% and 15.71% in G1 and with decreased cell number of 15.96% and 16.56% in S. Cell cycle related proteins CDK6 and CDC25A were down-regulated. The decreases of CDK6 and CDC25A by miR-449b were 39% and 22% respecyively; 49% and 32% by miR-34c respectively. The more decreases were seen in co-action by miR-449b and miR-34c with decreases of 69% in CDK6, 86% in CDC25A, and 59% in CyclinA., Conclusion: miR-449b and miR-34c resulted in cell cycle arrests and down-regulation of CDK6, CDC25A and CyclinA in high malignant ovarian cancer cell line SKOV3-ipl.

This study was aimed to clarify whether valproic acid (VPA) induces apoptosis of leukemia HL-60 cell line and its possible mechanism. The effect of different concentrations and treatment time of VPA on HL-60 cell proliferation was assayed by cytotoxicity test (CCK-8 method) and fluorescence microscopy, and flow cytometry was used to detect cell apoptosis. The expressions of telomerase subunit h-tert mRNA and apoptosis-related protein as well as caspase-3 activity were detected by real time-quantitative PCR, Western blot and ELISA respectively. The results indicated that VPA inhibited proliferation of HL-60 cells and induced cell apoptosis in a dose dependent manner (r = -0.87). The expressions of anti-apoptotic protein BCL-2 and h-tert mRNA were significantly decreased while the pro-apoptotic protein BAX and caspase-3 activity increased after treatment with VPA. The apoptosis rate of HL-60 cell was negatively correlated with expression of h-tert mRNA. It is concluded that VPA can inhibit leukemia HL-60 cell proliferation and induce apoptosis. The VPA displays anti-leukemia activity possibly through reducing h-tert mRNA and BCL-2 protein expression, increasing BAX expression and activity of caspase-3.

Objective: To investigate the effect of 3'-meisoindigo on the proliferation and the biological function of the splenocyte and thymocyte of mouse, which were 8 weeks old masculinity BALB/c, C57BL/6 and F1 hybridization mouse., Methods: Cells of thymus and spleen were harvested and prepared as the unicell suspension, then treated with 5, 10, 15, 20, 25 micromol/L 3'-meisoindigo. The cell proliferation was by MTT method, concentration of IL-12 was dectected by ELISA method, the mRNA levels of Bcl-2 and CDK2 were decected by RT-PCR. The cell cycle, apoptosis ratio, death ratio and intracellular ROS concentration were detected by FCM method. The protein level of Bcl-2, CDK2 and Bax were detected by immumofluorescence method., Results: 15, 20, 25 micromol/L 3'-meisoindigo can inhibit the proliferation of thymocyte and splenocyte (P < 0.05). It had dose-dependent and time-dependent manner. 3'-meisoindigo inhibit the secretion of IL-12, even at 5 micromol/L concentration. 15 micromol/L 3'-meisoindigo decrease the mRNA level of Bcl-2 and CDK2, induced apoptosis and G2 arrestting of the thymocyte and splenocyte. (P < 0.05). The intracellular ROS level increased after treated by 3'-meisoindigo at 15 micromol/L for 24 h (P < 0.05). There were no difference among three germ line mouse., Conclusion: Above 15 micromol/L, 3'-meisoindigo can inhibit the proliferation and externalization function of thymocyte and splenocyte from different germ line mouse, meanwhile the mRNA and protein level of Bcl-2 and CDK2 decrease, the Bax protein expressed increased, the intracellular ROS level increase too.

The study was aimed to investigate the expression of Toll-like receptor 4 (TLR4) on platelets and to determine whether platelet TLR4 involves in its activation induced by lipopolysaccharide (LPS). Human platelet-rich plasma (PRP) and platelet suspension obtained from 15 healthy individuals pretreated with a concentration of 0.2 microg/ml of LPS in the presence or absence of thrombin (1 U/ml) for 1 hour. The expressions of TLR4, CD62P (P-select) and CD40L on platelets were detected by flow cytometry, and platelet TLR4 expression was further determined by Western blot analysis. The results indicated that the percentage of TLR4-positive platelets induced by thrombin was increased by 32.34% compared with the resting platelets (25.44%, p < 0.05). TLR4 expression on platelets treated with LPS was remarkably elevated in the presence or absence of thrombin. However, the expression level of the former was much higher than that of the latter and thrombin stimulation alone (p < 0.05). Moreover, the similar results were found in Western blot analysis. Synchronously, expressions of CD62P and CD40L on resting platelets were 6.39% and 2.45%, they were also markedly increased when treated with thrombin (42.68% and 14.8%) and LPS respectively, and the increases of expression of CD62P and CD40L were more significant when stimulated with both LPS and thrombin (63.03% and 13.94%). Although anti-TLR4 antibody inhibited significantly the increase of TLR4, CD62P and CD40L on platelets induced by LPS, which did not affect their increase induced by thrombin. In conclusion, the evidence has been shown that functional TLR4 can be expressed on human platelets. It may involve in platelet activation as an important mediator of LPS-induced CD62P and CD40L expressions on platelets.

Sarcophagus beetles, which can not be replaced by Diptera, play a pivotal role not only in estimating PMI of dry human skeletal remains in the later stages decomposition of carcasses, but also the corruption, destruction, decomposition and posture changes of carcasses. This article explicates the succession of sarcophagus beetles on carrion and its influencing factors, and introduces the application and prospects of sarcophagus beetles in forensic entomology. Although few researches focus on sarcophagus beetles at present, it is believed that more and more forensic scientists will pay attention to sarcophagus beetles' application in forensic identification.

Objective: To compare different Mycobacterium tuberculosis antigens in enzyme-linked immunospot assay (ELISPOT) for the auxiliary diagnosis of active tuberculosis., Methods: The peripheral blood mononuclear cells (PBMC) from patients with tuberculosis and controls were co-cultured with the following antigens: purified protein derivative (PPD), early secretory antigenic target-6 (ESAT-6) and early secretory antigenic target-6/culture filtrate protein-10 fusion protein (ESAT-6/CFP-10). Spot forming cells (SFC) were enumerated by ELISPOT., Results: PPD-ELISPOT, E/C-ELISPOT and ESAT-6-ELISPOT showed significantly higher SFC counts in active tuberculosis [255(93-526), 148(40-354) and 28(10-116) respectively] as compared to the controls [10(5-41), 10(0-20) and 5(0-15) respectively], u values were 1479.5, 1390.5 and 2510.5 respectively, all P < 0.01. Compared with the sensitivity of ESAT-6-ELISPOT (62.1%), that of E/C-ELISPOT and PPD-ELISPOT was higher (90.3% and 84.8%), chi(2) = 17.496 and 28.541, all P < 0.01. Compared with the specificity of PPD-ELISPOT (68.9%), that of E/C-ELISPOT and ESAT-6-ELISPOT was also higher (84.4% and 88.9%), chi(2) = 6.807 and 10.808, P < 0.05 and P < 0.01 respectively., Conclusions: E/C-ELISPOT is a promising approach to the auxiliary diagnosis of tuberculosis, but its specificity could be affected by latent tuberculosis infection.

Objective: To compare enzyme-linked immunospot assay (ELISPOT) and tuberculin skin test (TST) and explore their roles in the auxiliary diagnosis of initial pulmonary tuberculosis., Methods: Totally 123 patients with initial pulmonary tuberculosis (tuberculosis group) and 102 patients with non-tuberculosis pulmonary disease (control group) were enrolled. The peripheral blood mononuclear cells of all participants were co-cultured with early secretiny antigen target-6/culture filtrate protein-10 fusion protein (ESAT-6/CFP-10), and spot forming cells (SFCs) were enumerated by ELISPOT (ESAT-6/CFP-10-ELISPOT). TST was also performed simultaneously., Results: ESAT-6/CFP-10-ELISPOT showed significantly higher numbers of SFCs after stimulation in tuberculosis group than in control group (P = 0.000). The sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, positive predictive value, and negative predictive value of ESAT-6/CFP-10-ELISPOT were 91.1% (111/123), 81.4% (82/102), 4.60, 0.12, 0.85, and 0.87 respectively, while the above values of TST were 65.6% (59/90), 45.1% (46/102), 1.31, 0.76, 0.51, and 0.60, respectively. The sensitivity and specificity of ESAT-6/CFP-10-ELISPOT were significantly higher than those of TST (all P = 0.000). The number of SFCs were not significantly different between smear-positive tuberculosis subgroup and smear-negative tuberculosis subgroup (P = 0.166). The sensitivities were 91.8% (67/73) and 88.0% (44/50) in these two subgroups, respectively, (P = 0.448)., Conclusions: ESAT-6/CFP-10-ELISPOT may be a more accurate approach for the auxiliary diagnosis of initial pulmonary tuberculosis; meanwhile, it offers certain diagnostic evidences for smear-negative tuberculosis. However, its specificity may be affected by latent tuberculosis infection. On the contrary, TST has poor value in the auxiliary diagnosis of initial pulmonary tuberculosis.

Objective: To examine global expression levels of microRNAs(miRNAs) in mouse cerebrum and to provide an important basis for detailed studies of individual miRNAs, their target genes, the miRNA-related regulatory networks in the mammalian central nervous system, and their implications in diseases., Methods: Low molecular weight RNA from cerebrum of five C57BL/6J mice were tailed and reverse transcribed by extended RT-primer. miRNA primers were carefully designed and arrayed on plates according to the Tm of each primer. PCR was carried out at different annealing temperatures using a gradient real-time PCR instrument. The relative expression level of each miRNA was calculated using 5sRNA for normalization., Results: Among the 285 miRNAs detected, 260 were positive with varying abundance. Their frequency distribution was approximately a normal distribution. The expression levels of most miRNAs were in accordance with previously published results by microarray. However, the positive rate was higher than that detected by microarray. miRNAs originating from the same hairpin precursors expressed at similar or significantly different levels. Clusters of proximal miRNAs were similar or quite different in abundance. It is suggested that the fate of miRNA after transcription determined their abundance., Conclusion: Using the RNA-tailing and primer-extension PCR array method, we obtained expression profile of miRNA in mouse cerebrum, especially the relative expression data of many low abundant miRNA in mouse cerebrum, which will be of special help for studying the fine-tuning function of low-level miRNAs.

Objective: To investigate the effect of valproic acid on the expression of P27(Kip1) and P170 and drug resistance of leukemia HL60/HT cell line and explore its possible mechanisms., Methods: HL-60/HT cells were derived from HL-60 cells induced by harringtonine (HT) in gradient concentrations. The inhibitory effect of valproic acid on the proliferation of HL-60 and HL-60/HT cells was evaluated by MTT assay, and the P27(Kip1) expression, P170 expression and cell cycle of the cells were analyzed with flow cytometry., Results: The multidrug-resistant HL-60/HT was acquired, which showed a stable drug-resistant index with increased IC(50) of HT, VCR, DNR and Ara-c by 9.30, 5.20, 4.91 and 3.65 folds, respectively, as compared with those of HL60 cells. The expression of P27(Kip1) in HL-60/HT cells was significantly lower but P170 expression significantly higher than that of HL-60 cells and normal mononuclear cells (P<0.05). The expressions of P27(Kip1) and P170 showed no significant difference between normal mononuclear cells and HL-60 cells. The growth inhibition rate of VPA combined with Ara-C was significantly higher than that of valproic acid or Ara-C alone in HL-60/HT cells and HL-60 cells (q=1.37 and 1.51, respectively). HL-60/HT and HL-60 cells cultured in the presence of VPA resulted in a significant increase in the expression of P27(Kip1) and the G(1)-phase cells (P<0.05), but the expression of P170 underwent no significant changes (P>0.05)., Conclusion: HL-60/HT cells have lower P27(Kip1) expression compared with HL-60 cells. Valproic acid can inhibit the growth of HL-60/HT cells and enhance their Ara-C sensitivity possibly by increasing P27(Kip1) expression and causing cell cycle arrest in G(1) phase.

Based on the continuous observation on located trees and branches, the phenology and sexual reproductive process of Xanthoceras sorbifolia Bunge in an annual growth cycle in Wudan area were investigated, and the relationships between fruit growth process of X. sorbifolia and effective accumulative temperature as well as the quantitative dynamics of flowers and fruits were studied. The results showed that the phenology of X. sorbifolia had a significant linear correlation with effective accumulative temperature, and there was a time sequence in the male flower silk elongating and antheral unfolding. The fruits of X. sorbifolia experienced three times of dropping during their growth process. The pollination rate was 26.4%, flower dropping rate was 73.6%, and fruit dropping rate was 92.9%, suggesting that the dropping of flowers and fruits was very serious. The fruits had a quicker vertical growth from early June to mid June and a quicker horizontal growth in the last ten-day of June, with the highest ratio of vertical to horizontal length being 2.17. The individuals of X. sorbifolia had definite differences in their phenology, which was disadvantageous to the cross pollination among the individuals, and the enhancement of pollination rate.

Objective: To study the protective effects of orientin against myocardial ischemia and hypoxia in rats., Methods: The protective effect of orientin against myocardial ischemia and hypoxia was observed in mice by recording their survival time under closed normobaric hypoxia and time of cardiac electric disappearance due to trachea clamping, in rabbits by evaluating arachidonic acid (AA)-induced blood platelet aggregation, in guinea pigs by measuring the coronal flow in the isolated heart and in SD rats with myocardial ischemia induced by pituitrin injection., Results: Orientin (1, 2, 4 mg/kg) significantly prolonged the survival time of mice under closed normobaric hypoxia and the gasping duration induced by decapitation. Orientin at concentrations of 3, 10, and 30 micromol/L also inhibited AA-induced blood platelet aggregation in rabbits and increased coronal flow in the isolated heart of guinea pigs. At 0.75, 1.5, and 3.0 mg/kg, orientin significantly antagonized pituitrin-induced ECG changes., Conclusion: Orientin may offer protection against myocardial ischemia and hypoxia in animal models in dose-dependent fashions.

Objective: To establish in vivo and in vitro models of persistent Mycobacterium tuberculosis infection, and therefore to study the persisters in different conditions and periods of chemotherapy, and to explore the relationship between the persisters and chemotherapy., Methods: The persisters in the two models were examined by culturing Mycobacterium tuberculosis in oxygen-starved condition and determining the mRNA expression of isocitrate lyase (ICL), alpha-crystallin chaperone (Acr) and 85B through quantitative PCR., Results: The bacteria which could be cultured in oxygen-starved condition were discovered in both models. The mRNA expression of ICL and Acr increased gradually and dramatically after culture for 4 days in the in vitro model, their values being (5.3 +/- 0.9) and (6.4 +/- 1.6) log copy/ml respectively. While the mRNA expression of 85B showed no significant change in oxygen-starved condition, it increased significantly in the standard condition, the values being (6.1 +/- 0.9) log copy/ml at 10th day. In the mice infected with Mycobacterium tuberculosis, the mRNA expression of ICL was detected in 2 and 4 weeks post-infection, and decreased 4 weeks after treatment. The Acr mRNA showed no or very low expression 4 weeks post-infection or 4 weeks after treatment, but it increased significantly 8 and 10 weeks after treatment, with a value of (6.2 +/- 1.7) log copy/ml at 10 weeks, and its expression was still detected 12 weeks after treatment and 4 weeks after the cessation of treatment [(3.0 +/- 1.6) log copy/ml]. The expression of 85B mRNA was high before the treatment [(6.4 +/- 1.1) log copy/ml], and decreased gradually during the therapy., Conclusion: The models of in vitro and in vivo persistence of Mycobacterium tuberculosis were established. ICL and Acr mRNAs were highly expressed, which may be the markers of persisters. Persisters can be detected by culture in oxygen-starved conditions and the measurement of mRNA expression.

Aim: To construct the recombinant eukaryotic expression vector pTARGET-TCR Vbeta8.2 and detect its their expression., Methods: Gene encoding TCR Vbeta8.2 was amplified by RT-PCR from spleen cells of Lewis rats, and then cloned into eukaryotic expression vector pTARGET. Recombinant clones were identified by blue/white screening on indicator plates after transformed into E.coli strain JM109, and then by bacteria colonies PCR and DNA sequencing. Recombinant plasmid was injected into BALB/c mice intramuscularly. Then the injected skeletal muscle was isolated, and expression of TCR Vbeta8.2 gene was detected by RT-PCR and immunohistochemistry. Immunocytochemical staining was used to detect the expression of pTARGET-TCR Vbeta8.2 gene after the recombinant plasmid was transfected into COS-7 cells by lipofectamine., Results: DNA sequencing demonstrated that TCR Vbeta8.2 gene was successfully inserted into pTARGET. RT-PCR demonstrated that TCR Vbeta;8.2 gene was successfully expressed in the injected muscle. Immunohistochemistry staining showed the expression of recombinant plasmid in the transfected COS-7 cells., Conclusion: The eukaryotic expression vector pTARGET-TCR Vbeta8.2 was successfully constructed and expressed in vivo and vitro, which would lay foundation for further studies on the protective effects of TCR Vbeta DNA vaccine on CIA.

Objective: To investigate the induction and culture of adventitious root of Panax notoginseng., Method: Three ways, induction from the explants of three-year-old P. notoginseng. The explants of regenerated shoots and calluses, were used to induce adventitious roots. The effects of 2, 4-dichlorophenoxyacetic acid, indole-3-butyric acid and naphthylacetic acid on adventitious root induction were investigated respectively. The effects of four modes of separating adventitious roots from the parent tissues on culture in vitro were compared., Result: Adventitious roots were successfully induced by three methods, of which the young flower bud callus was the best material for the induction of adventitious root. Indole-3-butyric acid possessed the strongest potency for induction. The liquid culture system was established by continuous culture of adventitious roots together with their parent tissues before separated., Conclusion: The acquisition and culture in vitro in liquid culture system of adventitious roots of P. notoginseng lay a foundation for the next investigation.

Objective: To evaluate the curative effect and safety of a long course regimen containing Chinese-made rifabutin as compared to the regimen containing rifapentine in the treatment of multi-drug resistant pulmonary tuberculosis., Method: During 18 month treatment, 130 patients with multi-drug resistant pulmonary tuberculosis were divided into a treatment group (rifabutin, pasiniazide, levofloxacin, ethambutol, ethionamide, amikacin for 3 months, rifabutin, pasiniazide, levofloxacin, ethambutol, ethionamide for 6 months, rifabutin, pasiniazide, levofloxacin, ethambutol for 9 months), and a control group (rifapentine, pasiniazide, levofloxacin, ethambutol, ethionamide, amikacin for 3 months, rifabutin, pasiniazide, levofloxacin, ethambutol, ethionamide for 6 months, rifabutin, pasiniazide, levofloxacin, ethambutol for 9 months) with proportion 1:1 random, and parallel compared method., Results: After intensive phase, the sputum negative conversion rates (smear negative, culture negative) of the treatment group and the control group were 41.54% (27/65) and 35.94% (23/65), chi(2) = 2.42, P > 0.05, respectively. The remarkable effective rates in chest X-ray of the two groups were all 10.77% (7/65), chi(2) = 0.01, P > 0.05, and the effective rates were 67.69% (44/65) and 56.92% (37/65), chi(2) = 1.44, P > 0.05, respectively. At the end of the treatment, the sputum negative conversion rate (smear negative, culture negative) of the treatment group was 75.0% (48/65), and of the control group was 65.08% (41/65), chi(2) = 1.88, P > 0.05. The remarkable effective rates in chest X-ray of the two groups were 46.15% (30/65) and 44.62% (29/65), chi(2) = 0.02, P > 0.05, and the effective rates were 76.92% (50/65) and 73.85% (48/65), chi(2) = 0.19, P > 0.05, respectively. The cavity closure rates were 23.64% (13/55) and 33.33% (17/51), chi(2) = 0.00, P > 0.05, respectively., Conclusion: Regimens containing rifabutin or rifapentine. are very effective in sputum negative conversion rate, lesion absorption and cavity closing for the treatment of multi-drug resistant pulmonary tuberculosis, with good safety and tolerance.

Objective: To investigate the expression level of genes located in chromosome 21 in the brain tissues of Down syndrome(DS)., Methods: An optimized semi-quantitative RT-PCR method was used to evaluate the expression levels of seven genes encoded in chromosome 21 in fetal cortex brain and cerebellum of DS and the control at the end of 20 weeks of gestation. B2M was used as internal reference to normalize cell loss., Results: The expression levels of 6 genes in cortex and cerebellum, including DYRK1A, SYNJ1, PCP4, C21orf5, C21orf2 and C21orf106, were comparable between DS and the control. ANA, a cell-cycle negative regulatory gene, was over-expressed dramatically in the cortex but not in the cerebellum of DS., Conclusion: Over-expression of ANA may contribute to the reduction of neuronal density in DS brain.

Objective: To investigate the changes of the goblet cells in the intestine during the restitution process of the gut barrier after hemorrhagic shock., Methods: Forty-nine Sprague-Dawley rats with body weight of 250-300 g were divided into control group (n=7) and experimental group (n=42). Rats in the experimental group was further divided into 6 groups (n=7 each) according to different time point at 1, 3, 6, 12, 24, and 36 hours after hemorrhagic shock resuscitation. The specimens from ileum tissue were taken to observe the morphological chan ges of the intestinal mucosa. The number of goblet cells was determined by light microscope and/or electron microscope. The contents of trefoil factor family 3 (TFF3) of goblet cells were examined using GC-9A gas chromatographic instrument., Results: After hemorrhagic shock, mucosal epithelial injury was obvious in the small intestine. Tissue restitution was found after 3 hours, and mostly established after 12 hours. Following tissue restitution,the denuded mucosal surface was covered intensively by goblet cells. The number of goblet cells on the intestinal mucosa was reduced significantly from 243+/- 13 at 1 h to 157+/- 9 at 24 h (r=- 0.910, P< 0.01), and returned to normal level at 36 h. In the experimental group, the content of TFF3 in the intestinal mucosa increased significantly at 12 hours, decreased, but was still higher at 24 hours (t=3.24, P< 0.05)., Conclusions: The goblet cells play a key role in the restitution of intestinal mucosa. High expression of TFF3 may facilitate the intestinal mucosal restitution in the early phase.

Objective: To observe the effects of pravastatin on platelet-derived nitric oxide system in hypercholesterolemia (HC) and atherosclerosis (AS) in rabbits, and the relationship between these changes and atherosclerosis courses., Methods: Thirty male New Zealand white rabbits were randomly divided into three groups, 12 in group A, 12 in group B, and 6 in group C. All of them were fed daily with cholesterol-rich food during the first 12 weeks. In addition, in group A, pravastatin (10 mg) was orally administered daily. At the end of the 12th week, 6 in group A and B were killed randomly and their aortas were removed and the pathologic changes were observed. In the following 12 weeks, food enriched with cholesterol was substituted with normal food in all three groups. Pravastatin treatment was continued or started in the remaining members of group A and group B, but not in group C. At the end 24th week, all rabbits were killed and their aortas were examined for the fatty-streaks or atherosclerotic plaques. The expressions of endothelial NOS (eNOS) mRNA and inducible NOS (iNOS ) mRNA, NOS activity, NO production and the level of the serum lipids were measured at 0, 6th, 12th, 18th and 24th week., Results: The expression levels of platelet-derived NOS mRNA, eNOS mRNA ratio in group A had no difference at above time points, while in group B were reduced significantly at 6th week and 12th week compared with at 0 week (P <0.01), and increased at 18th week and 24th week compared with 12th week (P <0.05). The expression levels of eNOS mRNA in group C were reduced at 6th, 12th and 18th, 24th week compared with 0 week (P <0.05 and P <0.01, respectively), and were reduced in groups B and C compared with group A at 6th ,12th week (P < 0.05) and increased in group A and B compared with group C at 18th, 24th week (P <0.01). The expression levels of iNOS/mRNA among the three groups had no difference. Pathologic finding of the arteries: AS was not found in group A from the 12th to 24th week. While in group B, there were a lot of fatty-streaks on the entire intima of all large arteries at the 12th week. There were also fatty-streaks in the ascending aorta, but were improved at the 24th week. In group C, there were marked plaques in the entire aorta at the 24th week., Conclusions: The expressions of platelet-derived eNOS mRNA, NOS activity, NO production are decreased in HC or AS rabbits. Pravastatin can up-regulate expressions of platelet-derived eNOS mRNA, NOS activity, leading to preventing or improving the pathological courses of AS.

Objective: To explore the effects of chloride channels on the regulation of platelet cytoplasmic free calcium concentration ([Ca2+]i) and platelet aggregation (PAG)., Methods: Freshly separated platelets were activated by thrombin. Chloride channel blockers DIDS or NFA and calcium channel blockers SK&F96365 or nifedipine were added to study the effects on platelet [Ca2+]i and PAG by a single reagent or the combination of reagents and find out the interactions among DIDS, NFA, SK&F96365 and nifedipine., Results: Both DIDS and NFA could inhibit the thrombin (1 U/ml) induced PAG in a dose-dependent manner, whereas had little effect on resting [Ca2+]i. As compared with the control group, DIDS, SK&F96365 and Nifedipine could significantly reduce the PAG, Ca2+ release and Ca2+ influx in thrombin activated platelet (P < 0.05). The combination of DIDS and SK&F96365 had greater effects in reducing the PAG, Ca2+ release and Ca2+ influx than either reagent alone (P < 0.05). The combination of DIDS and nifedipine also had greater effect than each alone in reducing Ca2+ release (P < 0.05). The combination of NFA and SK&F96365 weakened each other's effect on Ca2+ release (P < 0.05), while NFA and nifedipine weakened each other's effects on PAG, Ca2+ release and Ca2+ influx in thrombin activated platelet (P < 0.05)., Conclusion: DIDS and NFA have no effect on the resting [Ca2+]i and the leak calcium influx of platelet. DIDS can inhibit the Ca2+ release, Ca2+ influx and PAG of platelet induced by thrombin, while NFA can only inhibit the Ca2+ release. The chloride channel and calcium channel blockers have interactions in affecting resting [Ca2+]i and PAG of platelet. The opening of chloride channel can influence the cellular calcium movement of platelet.

Objective: To investigate the platelet activity and function in pregnancy induced hypertension (PIH) and gestational diabetes mellitus (GDM)., Methods: Twenty-one patients with GDM and 23 patients with PIH in third-trimester were included. Twenty normal pregnant women in third-trimester served as controls. Platelet count (PC), mean platelet volume (MPV) were determined on Cell-DYN 1600 and the expression of CD62P was analyzed on FACSC alibur., Results: (1) PC was (181 +/- 56) x 10(9)/L in PIH, (206 +/- 60) x 10(9)/L in GDM and (229 +/- 56) x 10(9)/L in controls, respectively. PC in PIH was lower than that of controls (P < 0.01), but there was no significant difference between GDM and controls. (2) MPV was (11.2 +/- 2.0) fl in PIH, significantly higher than that of controls (8.7 +/- 1.6) fl (P < 0.001). In GDM, MPV was (9.5 +/- 1.6) fl, without significant difference compared with that of controls. (3) The expression of CD62P increased significantly in PIH compared with controls [CD62P: (42 +/- 13)% vs (26 +/- 7)%, P < 0.001; CD62P(I): 109 +/- 39 vs 75 +/- 13, P < 0.01]. In GDM, the expression of CD62P also increased significantly compared with the normal pregnancy [CD62P(%): (42 +/- 14)% vs (26 +/- 7), P < 0.001; CD62P(I): 100 +/- 42 vs 75 +/- 13, P < 0.05]. (4) All parameters had no significant difference between PIH and GDM., Conclusion: Platelet activity is enhanced in PIH and GDM. It may play an important role in the pathogenesis and development of the two diseases.

Objective: To study the immunotherapeutic effect on the esophageal adenocarcinoma mediated by gp96-peptide complexes isolated from the same kind of tumor., Methods: gp96-peptide complexes were purified from nude mice tumors burdened by subcutaneous injection of human esophageal adenocarcinoma cell line SEG-1. gp96-peptide complexes were carried by the dendritic cells(DC) induced from human peripheral blood mononuclear cells to prepare gp96-DC vaccine. The proliferation of lymphocytes was tested with trypan-blue stain. The quantity of interferon-gamma(IFN-gamma) released from cytotoxic T lymphocytes (CTL) was detected with ELISA method. The killing effect of CTL on target cell SEG-1 was measured with MTT., Results: We obtained 120 microg gp96 from 55 g tumor tissue. DC, gp96, and gp96-DC all could elicit the proliferation of lymphocytes and make them becoming into CTL which released IFN-gamma and showed different degrees of killing effect on target cell SEG-1. gp96-DC has the strongest eliciting effect among them. At the ratio of E(effect) to T(target) as 40:1,the killing rate was 68%. No significant difference between the effects of CTL induced by DC alone and of lymphocytes without specific antigen on SEG-1 and K562 cells., Conclusion: The gp96-peptide complexes from tumors can improve the effect of eliciting lymphocyte proliferation of DC and make the lymphocyte becoming into CTL more effectively. These CTLs show prominent killing effect on the target tumor cells.

Objective: To explore the effects of 2A-1-1 (purified component from Panax notoginsengs saponins) on the aggregation of and Ca2+ influx into human platelets., Methods: The aggregation of platelets was tested by nephelometry, Fura-2 fluorescent technique was used for detecting cell [Ca2+]i. The effects of 2A-1-1, nifedipine and SK&F96365 on Ca(2+) influx into human platelets induced by ADP or CPA were observed separately., Results: Nifedipine (< 20 micromol/L) could not inhibit platelet aggregation induced by ADP or the Ca(2+) influx induced by ADP or CPA. SK&F96365 at 20 micromol/L could inhibit the maximal aggregation of platelets induced by ADP with a inhibitory rate of 59.83%, at 15 micromol/L could inhibit the Ca2+ influx induced by CPA or ADP. 2A-1-1 (5, 10 and 20 micromol/L) could inhibit the maximal aggregation of platelets induced by ADP with the inhibitory rates of 47.06%, 53.47% and 71.52%, respectively. 2A-1-1 at 10 and 20 micromol/L could inhibit the Ca2+ influx induced by CPA or ADP., Conclusions: 2A-1-1 can inhibit platelets aggregation, block the ROC (Receptor-dependent Ca2+ channels) and inhibit Ca2+ influx of human platelets.

Objectives: To study on the changes of intracellular calcium and magnesium in cirrhosis and its clinical significance., Methods: The calcium and magnesium were determined in serum (SCa, SMg), platelets (PCa, PMg), mononuclear cells (MNCCa, MNCMg), polymorphonuclear cells (PMNCa, PMNMg) and erythrocytes (RCa, RMg) of 50 patients with uncompensative cirrhosis (group A) and of 35 patients with compensative cirrhosis (group B). 35 health persons were the control group., Results: The SCa and SMg of group A were lower significantly than those of both group B and control group. The MNCCa, PMNCa, RCa, PMg, MNCMg, PMNMg, RMg of group A [(4.76+/-1.91) micromol/10(9), (7.56+/-2.88) micromol/10(9), (0.66+/-0.13) mmol/L, (5.53+/-2.25) micromol/10(11), (6.64+/-3.53) micromol/10(9), (10.12+/-4.32) micromol/10(9), (2.02+/-0.76) mmol/L] and those of group B [(5.34+/-2.41) micromol/10(9), (8.32+/-2.34) micromol/10(9), (0.67+/-0.11) mmol/L, (5.55+/-2.67) micromol/10(11), (6.56+/-3.44) micromol/10(9), (10.95+/-4.45) micromol/10(9), (2.21+/-0.74) mmol/L] were lower significantly than those of control group [(6.86+/-2.02) micromol/10(9), (9.89+/-3.23) micromol/10(9), (0.72+/-0.10) mmol/L, (7.43+/-2.78) micromol/10(11), (8.68+/-4.1) micromol/10(9), (13.96+/-5.76) micromol/10(9), (2.74+/-0.92) mmol/L]; t (group A vs. control group)=4.88, 3.48, 2.31, 3.45, 2.46, 3.52, 4.00, 0.01, 0.01, 0.05, 0.01, 0.02, 0.01, 0.01; t (group B vs. control group)=2.87, 2.34, 2.00, 2.89, 2.33, 2.45, 2.65, 0.01, 0.05, 0.05, 0.01, 0.05, 0.02, 0.02. The PCa of the patients with hepatic encephalopathy was higher, the SMg, PMg, MNCMg, PMNMg and RMg were lower than those of the patients without hepatic encephalopathy significantly. The SCa, SMg, PMg, MNCMg, PMNMg and RMg of the patients in Child stage C were lower significantly than those of the patients in Child stage B. There were no significant differences of PCa, MNCCa, PMNCa and RCa between Child stage C and Child stage B. There were no significant differences of SCa, MNCCa, PMNCa and RCa between the patients with and without hepatic encephalopathy. The ratios of PCa/SCa, MNCCa/SCa and PMNCa/SCa of the patients with decreased SMg were lower than those of control group. The SMg, MNCMg, PMNMg and RMg were correlated directly with the level of serum albumin., Conclusion: There are calcium and magnesium deficiencies in the patients with uncompensative cirrhosis and compensative cirrhosis, this deficiency aggravates with the severity of the disease. There is relative increase of intracellular calcium. The magnesium deficiency may be one of the reasons for both hepatic encephalopathy and relative increase of intracellular calcium.

Objective: To investigate measures to prevent the outbreak of severe acute respiratory syndrome (SARS) in healthcare workers in isolation units., Methods: The architectural factors and the infection of healthcare workers in different wards in our hospital between 30 January 2003 and 30 March 2003 were analyzed., Results: Four kinds of isolation wards were evaluated, including the ward where the thirty-first bed lied in on the twelfth floor, the laminar flow ward in the intensive care unit (ICU) where the tenth bed lied in on the fifteenth floor, the ward where the twenty-seventh bed lied in on the thirteenth floor of Building A, and thirty wards on the fourteenth to eighteenth floors of Building B. The ratios (m2/m3) of the area of the ventilation windows to the volume of the room were 0, 0, 1:95 and 1:40, respectively. Numbers of SARS cases in the wards mentioned above were 1, 1, 1 and 96, respectively. The total lengths (hour) of hospitalization were 43, 168, 110 and 1,272, respectively. The infection rates of the healthcare workers in the areas mentioned above were 73%, 32%, 28% and 2%, respectively. The difference of the infection rates was of statistical significance., Conclusion: In addition to strict personal protective measures, isolation of SARS cases in wards with high ratio of the area of ventilation windows to the volume of the room and good ventilation may be the key to preventing the outbreak of SARS in healthcare workers in isolation units.

Objective: To study and evaluate the efficacy and safety of recombinant human interleukin-2 (IL-2) in the treatment of pulmonary tuberculosis., Methods: Two hundred and nine cases with re-treated Mycobacterium tuberculosis-positive pulmonary tuberculosis were randomly divided into a trial group (106 cases, treated with 3PaZ (TH)L(2)VE(AK) + IL-2/4PaL(2)V) and a control group (103 cases, treated with 3PaZ(TH)L(2)VE(AK)/4PaL(2)V). The efficacy of 203 cases was available for evaluation when the course was completed (trial group 103 cases, control group 100 cases)., Results: The sputum smear-negative conversion rates at the 1st and the 2nd month of therapy were 33.3% and 69.4% in the trial group, and 7.2% and 44.9% in the control group (P < 0.01). At the completion of the therapy, the X-ray resolution rates were 64.1% and 36.0% respectively for the trial and the control groups, the difference being significant (P < 0.001). There were significant differences in CD(4) T cells, the ratio of CD(4)/CD(8) and NK cells between the two groups (P < 0.01). The level of soluble interleukin-2 receptor (sIL-2R) was significantly different between the two groups after treatment for 3 months (P < 0.05). IL-2 associated side effects were rare and mild., Conclusion: As an effective and relatively safe biological agent, IL-2 can be added to the standard chemotherapy for pulmonary tuberculosis.

Objective: To describe a hospital outbreak of severe acute respiratory syndrome (SARS) and summarize the clinical features and therapeutic approaches., Methods: Clinical data in this cohort were collected prospectively as they were identified., Results: The outbreak started with a SARS patient from the community on 30 January 2003, followed by a total of 96 people [76 women and 20 men; mean age (29.5 +/- 10.3) years; 93.8% of whom were health care workers] infected in a short period of time after their exposure to this source patient. The incubation period ranged from 1 to 20 days, with a mean of (5.9 +/- 3.5) days. The initial temperature was (38.3 +/- 0.6) degrees C, while the highest was (39.2 +/- 0.6) degrees C (P < 0.001), with a mean fever duration of (9.0 +/- 4.2) days. Other common symptoms included fatigue, cough, mild sputum production, chills, headache, general malaise and myalgia. The radiographic changes were predominantly bilateral and in the middle or lower lung zones. Leukopenia was observed in 67.7% of this cohort. The mean lowest oxygen saturation was (94.8 +/- 3.1)% with supplementary oxygen through a nasal cannula. 68.8% of the patients were treated with methylprednisolone for a mean period l of (4.9 +/- 2.4) days. The initial dose was (67.3 +/- 28.2) mg/d and the maximal dose was (82.4 +/- 30.5) mg/d. Ninety-five patients (99.0%) had a complete clinical recovery, and 1 patient died of progressive acute respiratory distress syndrome. The mean hospitalized duration was (17.2 +/- 8.0) days., Conclusion: SARS appears to be highly contagious and potentially lethal among health care workers, characterized by acute onset and rapid progression. Corticosteroids, antibiotics, human gamma-globulin, interferon-alpha, and antiviral drugs, although used empirically, might be of some benefits in shortening the clinical course.

To establish a quantitative assay for telomerase activity and analyze the telomerase activity in peripheral blood mononuclear cells (PBMNC) from patients with acute leukemia, a fluorescent dye, PicoGreen, was added to the products after telomere repeat amplification protocol. The samples were excited at 480 nm and the fluorescence emission intensity was measured at 520 nm using a spectrofluorometer. Telomerase activity was detected in PBMNCs from 20 cases of normal individuals and 25 patients with acute leukemia. The results showed that the fluorescence of PicoGreen binding to double-stranded DNA specifically was enhanced with increase of DNA quantities. In conclusion, the met hod is rapid, simple and quantitative, the telomerase activities of PBMNCs from acute leukemia patients are significantly higher than that of the normal controls.