This paper reviews recent development and applications of high-speed countercurrent chromatography (HSCCC) in the separations on the analytical scale,the semi-preparative and preparative scale of natural products,antibiotics,proteins,inorganic compounds etc.Several series of two-phase solvent systems to facilitate the systematic search suitable for HSCCC are introduced.And the new countercurrent chromatography (CCC)technologies such as HSCCC coupled with MS,pH-zone-refining countercurrent chromatography,and ion-pairing Countercurrent Chromatography are explained and prospected in term of applications.

Objective: To introduce a new way, which facilitates the systematic search and evaluation of the solvent systems suitable for high-speed countercurrent chromatography (HSCCC). Methods: The software integrated with the thermodynamics solution functions and theory is directive to calculate the composition, physical parameters, such as volume, viscosity, dielectric constants, and polarity parameters of the upper and lower phases. Results: Separation of Guizhi-decoction Fr. A by HSCCC, one of traditional Chinese medcines, is conducted to prove the applicability of this software in selection and optimization of solvent systems. Conclusion: This software can be used to select and optimize the solvent systems of HSCCC. It is supposed to extend this method.

Objective To investigate the effects ofZhenqi-Fuzheng capsule combined with highly active antiretroviral therapy (HAART) the Thl/Th2, Tcl/Tc2, and CD4+ T lymphocytes in patients with advanced AIDS.Methods A total of 70 patients with advanced AIDS were randomly divided into a treatment group and a control group, with 35 patients in each group. The treatment group was treated withZhenqi-Fuzheng capsule combined with HAART, while the control group was treated with HAART only. The patients in both groups were treated for 24 weeks. The Thl, Th2, Tcl, Tc2 and CD4+ T lymphocytes in peripheral blood were detected by flow cytometer.Results After treatment, the ratios of Th1/Th2 (6.27 ± 0.70vs. 5.39 ± 0.64;P<0.05) and Tcl/Tc2 (10.70 ± 0.68vs. 10.16 ± 0.85;P<0.05) in the treatment group showed significantly higher than those in the control group. The CD4+T lymphocytes after the treatment in both groups showed significantly increased than that before the treatment (treatment group: 289.2 ± 101.5/mlvs. 121.1 ± 77.5/ml,P<0.01; control group: 231.6 ± 97.6/mlvs. 118.9 ± 76.2/ml,P<0.01). The CD4+T lymphocytes in the treatment group showed significantly increased than that in the control group after the treatment (P<0.05).ConclusionsZhenqi-Fuzhengcapsule combined with HAART may regulate the Thl/Th2 and Tcl/Tc2, and increase the CD4+ T lymphocytes in patients with advanced AIDS.

Objective To explore clinical effect of small dose of glucocorticoid in the treatment of AECOPD patients with severe disease associated with low levels of cortisol.Methods72 patients with AECOPD in our hospital were randomly divided into control group and treatment group.The control group was treated with conventional AECOPD method,the treatment group was treated on the base of the treatment group with a small dose of corticosteroids,CRP, IL-6, TNF-α, FEV1, PCT and other indicators, as well as patients with ventilation, shock and mortality were compared after the end of treatment.ResultsThe time of non mechanical ventilation, the time of non-mechanical ventilation, and the time of non-shock were increased, the shock and the mortality was lower in the treatment group,(P<0.05);The CRP level of patients in the treatment group was lower(P<0.05);IL-6, TNF-α, PCT levels of patients in the treatment group was lower (P<0.05);FEV1, FVC and FEV1/FVC level in the treatment group were higher(P<0.05).ConclusionSmall dose glucocorticoid can improve the pulmonary function of patients with AECOPD complicated by severe disease, and alleviate the disease, significantly reduce the levels of inflammatory factors, promote the improvement of the lung function.

Due to an overall high incidence of pulmonary tuberculosis (TB) and the emergence of drug-resistant TB, the role of surgical treatment is likely to be expanding. This review discusses the use of surgery in the treatment of TB, including surgical indications, timing of surgery and preoperative management, type of operations, and postoperative anti-TB treatment.

SHIV-CN97001 played an important role in assessing the immune effect and strategy of the AIDS vaccine which included genes of the predominant prevalent HIV-1 strain in China. In this study, SHIV-CN97001 was in vivo passaged serially to construct pathogenic SHIV-CN97001/rhesus macaques model. To identify variation in the gp120 region of SHIV-CN97001 during passage, the fragments of gp120 gene were amplified by RT-PCR from the plasma of SHIV-CN97001 infected animals at the peak viral load time point and the gene distances (divergence, diversity) were calculated using DISTANCE. The analysis revealed that the genetic distances of SHIV-CN97001 in the third passage animals were the highest during in vivo passage. It had a relationship between viral divergence from the founder strain and viral replication ability. The nucleic acid sequence of the V3 region was highly conservative. All of the SHIV-CN97001 strains had V3 loop central motif (GPGQ) and were predicted to be using CCR5 co-receptor on the basis of the critical amino acids within V3 loop. These results show that there was no significant increase in the genetic distance during serial passage, and SHIV-CN97001 gp120 gene evolved toward ancestral states upon transmission to a new host. This could partly explain why there was no pathogenic viral strain obtained during in vivo passage.

Objective To investigate the diagnostic value of macrophage migration inhibitory factor (MIF) for hepatocellular carcinoma (HCC).MethodsThe research was divided into 2 parts,including testing research and confirmatory research.The clinical data of 269 patients with HCC ( group A) and 390 individuals (including 135 patients with hepatic cirrhosis,106 with benign hepatic diseases and 149 healthy individuals,control group A) who were admitted to the Zhongshan Hospital of Fudan University from January to May,2004,and 173 patients with hepatic cancer (group B) and 257 individuals (including 86 patients with hepatic cirrhosis,79 with benign hepatic diseases and 92 healthy individuals,control group B ) who were admitted from August to December,2004,and 80 patients with HCC who received radical hepatic resection in January 2005 were retrospectively analyzed.Samples of plasma of patients in the group A and individuals in the control group A were collected before operation.Samples of plasma of patients received radical hepatic resection were collected preoperatively and at postoperative day 3,7 and 30.HCC and adjacent issues of patients in the group A were collected.The levels of MIF in the plasma and tissues were detected by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry,respectively.Non-normal distribution data were described as M( QR).Differences between the groups were analyzed by using the Mann-Whitney U test,and the relationship between the levels of MIF in the plasma and tissues was detected by the Spearman correlation coefficient.The diagnostic value of MIF was analyzed by the ROC curve.ResultsThe levels of MIF in the plasma of patients in the group A and individuals in the control group A were 85.7 μg/L (58.8 μg/L) and 15.5 μg/L(31.6 μg/L),respectively.The levels of MIF in the plasma of the patients with hepatic cirrhosis,benign hepatic diseases and healthy individuals were 24.9 μg/L (12.6 μg/L),12.5 μg/L(7.3 μg/L) and 13.2 μg/L (7.7 μg/L),respectively.There was a significant difference in the level of MIF between the group A and the control group A (F =54.235,P ＜ 0.05 ).The area under the ROC curve reached peak when the level of MIF in the plasma was 35.3μg/L.Compared with the control group B,the vdues of AUC,sensitivity and specificity were 92.1％,90.7％ and 93.4％ in the group B.The levels of MIF of the patients with HCC before operation and at 3,7,and 30 days after operation were 81.0 μg/L(54.0 μg/L),76.1 μg/L(47.5 μg/L),50.9 μg/L (40.7 μg/L) and 18.7 μg/L ( 15.1 μg/L),respectively.The levels of MIF decreased with time passed by,and were back to normal at 30 days after the operation.The median expressions of MIF in the HCC and adjacent issues were 0.083 and 0.007,respectively,with a significant difference ( U =3.975,P ＜ 0.05).The expression of MIF in the plasma was positively correlated with its expression in the HCC tissue ( r =0.759,P ＜ 0.05 ).ConclusionMIF plays an important role in the genesis and development of HCC and has potential to be one of the molecular markers for the diagnosis of HCC.

Objective To explore the synergetic effect of HBX protein and M2 macrophages in inflammatory microenvironment on invasion and metastasis of hepatocellular carcinoma cells.Methods Hep3B cells were infected with recombinant lentivirus carrying HBx gene,following co-culture with THP-1 original M2 macrophages.The cells were divided into six groups:two infected groups (Hep3B +and Hep3B + + M2),four non-infected groups (Hep3B-,Hep3B-+ LV5,Hep3B-+ M2,Hep3B-+LV5 + M2).Western blot (WB) was used to assess the expression changes of E-cadherin and N-cadherin,markers of epithelial-mesenchymal transition (EMT).The cellular location of EMT markers was observed by immunofluorescence confocal microscopy.Transwell assay was used to evaluate the invasion ability of Hep3B cells.Results HBX protein overexpressed in Hep3B cells by lentivirus infection.After 72 h co-culture with M2 macrophages,WB results showed that E-cadherin descreased significantly in Hep3B+ (0.42 ±0.11) when compared with Hep3B-(1.00 ±0.18) (t =4.762,P ＜0.05),while N-cadherin was significantly higher in Hep3B + (2.85 ± 0.44) than in Hep3B-(1.00 ± 0.17) (t =4.762,P ＜ 0.05).M2macrophages decreased E-cadherin expression in Hep3 B + + M2 (0.1 ± 0.13) compared with Hep3 B + (t =3.255,P ＜0.05),while N-cadherin expression increased in Hep3B+ + M2 (4.18 ± 0.52) (t=10.009,P ＜ 0.05).Non-Infected groups didn't change the markers of E-cadherin and N-cadherin.It was suggested that invasion ability of Hep3B increased by HBx overexpression.Conclusions HBX protein and M2 macrophages synergetically mediated the invasion and metastasis of hepatocellular carcinoma cells by EMT.

Objective By analyzing the expression of cluster of differentiation 74 (CD74) in hepatocellular carcinoma (HCC) and HCC cell lines,the correlation between the level of CD74 expression and the patients' prognosis was investigated.MethodsThe expression of CD74 in high metastatic potential HCC cell lines(MHCC-LM3,MHCC-97H),low metastatic potential HCC cell line( MHCC-97L),and no metastatic potential HCC cell line(Hep-G2) were estimated by Western blot.The paraffin embedded tissues which include intra-tumor and paratumor tissues were collected from 320 patients who had received HCC curative surgical resection and 5 normal liver tissues from the donors of liver tranplantation.The high density tissue micro-array was made of these specimens. Immunol-histochemistry was applied to discover the different levels of CD74 in tumor,paratumor and normal liver tissues.Survival curves were generated by the Kaplan-Meier method and verified by the Logrank test.Cox proportional hazards regression analysis was applied to estimate the prognostic factors in multivariate analysis.ResultsThe expression level of CD74 was significantly higher in low metastatic potential and no metastatic potential HCC cell lines (MHCC-97L 1.224 ±0.014,Hep-G2 1.374 ±0.006) than that in high metastatic potential ones( MHCC-LM3 0.622 ±0.078,MHCC-97H 0.732 ± 0.083 ).Significant differences can be found between the groups (t =- 13.308,- 16.849,- 10.177,- 13.436,- 17.057; P ＜0.01 ).Meanwhile,in tumor tissues,the CD74 was expressed positively in 221 patients and negatively in 99 patients.But CD74 was expressed slightly in paratumor and negatively in 5 normal liver tissues.There's no significant differences between the groups categorization according to age,HBsAg,cirrhosis,AFP level,tumor number,tumor size,tumor capsule,blood vessel invasion,Edmondson Grades and tumor nodes metastasis classification (TNM) stages (x2 =0.053,0.141,1.200,0.000,0.277,1.975,0.263,1.044,0.000,0.433 ; P ＞ 0.05 ),except gender (x2 =3.954,P ＜ 0.05).Kaplan-Meier method showed that patients with positively expression of CD74 had better prognosis than others (x2 =5.620,P ＜ 0.05 ).Cox proportional hazards regression analysis showed that CD74 was a significant and independent prognostic factor of survival [ hazard ratio (HR) =0.721,95％confidence interval (CI) =0.522 - 0.996,P ＜ 0.05 ].Conclusion The expression of CD74 in hepatocellular carcinoma could be a biomarker of the prognosis and there's some potential correlation with cancer cell apoptosis.

Objective:To investigate the expression of circhipk3 in microglial cells in heat-induced neurological injury, and to preliminary analyze the effect of circhipk3 on microglial polarization in heat-induced neurological injury.Methods:Mice were randomly (random number) divided into a control group and a heat radiation disease 0.8 h group (HS 0.8), a heat radiation disease 8h group (HS 8), and a heat radiation disease 24 h group (HS 24). By establishing a mouse model of heat shock (HS), heat-damaged brain tissue was obtained, microglia were isolated and RNA was extracted. Quantitative PCR method was used to detect M1 and M2 marker molecules in microglia, and to evaluate the polarization direction and type of microglia. The expression level of circhipk3 was detected in microglial cells in heat-induced neurological injury, and the effect of circhipk3 on microglial polarization was further elucidated by intervening the expression of circhipk3 in microglial cells.Results:The expression of CD45 and CD11-b in the HS 8 group was significantly higher than that in the control group [(4.41±0.18) vs. (1±0.15), P=0.000], [(3.47±0.19) vs (1±0.15), P=0.000] , and the CD45 and CD11-b of the HS 24 group was significantly lower than that of the HS 8 group [(1.34±0.15) vs. (4.41±0.18), P=0.000], [(1.38±0.21) vs. (3.47±0.19), P= 0.001]. At the same time, the expression of CD206, FIZZ and Arg1 in the HS 8 group started to increase compared with the control group [(1.59±0.16) vs. (1±0.12), P=0.014], [(1.62±0.15) vs. (1±0.15), P=0.002 ], [(2.23±0.28) vs. (1±0.19), P=0.004], and CD206, FIZZ, and Arg1 in the HS 24 group were significantly higher than those in the control group [(2.67±0.20) vs. (1±0.12), P=0.002], [(2.19±0.15) vs. (1±0.15), P=0.000], [(3.04±0.18) vs. (1±0.19), P=0.001]; circhipk3 mimicis significantly increased the expression of Arg1 [(7.26± 0.06) vs. (3.86±0.06), P=0.000]; at the same time, circhipk3 inhibitor promoted the expression of CD45 and HO-1 [(2.96±0.03) vs. (1.63±0.09), P=0.000], [(2.52±0.10) vs. ( 1.30±0.02), P=0.000]. Conclusions:Microglial cells are predominantly M1-type in early neurological injury of heat radiation disease. HO-1 may be one of the microglial M1-type markers. The high expression of circhipk3 in microglial cells mainly promotes its transformation to M2 type.