The aim of laboratory evaluation of basic education is to improve the quality of experimental teaching,which demands thorough reforms of many aspects,including the ranks of teachers,administration,content,methods and instrument of teaching. This article summarizes the experience and achievement of Functional Laboratory Centre,Central South University during under- graduate teaching evaluation by the Ministry of Education in 2006.

Objective:To investigate the quantification of CD4~+CD25~+ regulatory T cells and distribution of CD4~+CD8~+ T lymphocyte subgroup in peripheral blood of patients in chronic hepatitis B (CHB),and to reveal relationship between CD4~+CD25~+ regulatory T cells,CD4~+CD8~+ T lymphocyte subgroup and HBV infetion as well.Methods:CD4~+CD25~(high),CD4~+CD25~+Foxp3~+Treg and CD3~+CD4~+CD8~+T lymphocyte subgroup in peripheral blood from 50 patients with CHB and 20 healthy controls was analyzed using flow cytometry.HBV DNA was detected by fluorescence quantitative PCR.Results:The number of CD4~+CD25~(high)Tregs in patients with CHB was obviously higher than that in healthy controls(P＜0.01)and increased with copies of HBV DNA.The same with the change of CD4~+CD25~+Foxp3~+Tregs in patients with CHB and there was a positive correlation between CD4~+CD25~(high)Tregs and CD4~+CD25~+Foxp3~+Tregs(r=0.890,P＜0.001).Compared with healthy controls,the frequency of CD4~+T cells and the ratio of CD4~+/CD8~+ in patients with CHB was declined,but there was no significant difference in the frequency of CD3~+T cells and CD8~+T cells between them(P＞0.05).The variation in the number of CD4~+CD25~(high)Tregs was correlated positively with the copies of HBV DNA(r=0.782,P＜0.001)and glutamic-pyruvic transaminase(ALT)(r=0.432,P＜0.005)separately,but negatively with the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+(P＞0.05).The variation in the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+ was also correlated negatively with the copies of HBV DNA(P＞0.05).Conclusion:The number of CD4~+CD25~(high)Tregs increases in patients with CHB and is in accordance with the copies of HBV DNA and increased level of ALT.Further studies should be done to investigate weather CD4~+CD8~+ T lymphocyte subgroup could be used to monitor the state of community.

Objective To investigate the relationship between the steroid-sensitive nephrotic syndrome(SSNS) and interleukin-18(IL-18) and to approach the inhibitive role of dexamethasone(DEX) on expression of IL-18 of peripheral blood mononuclear cells(PBMC) in children with SSNS in vitro.Methods IL-18 levels of serum, urine and supernatants of PBMC cultured in vitro were measured by enzyme linked immunosorbent assay(ELISA) in 23 children with SSNS who were either before or after treatment. Fifteen age-matched healthy children served as normal control group, and another 18 children with respiratory infections as infectious control group.Results There were signi-ficant differences of IL-18 in serum and urine before and after treatment in children with SSNS (t=15.072,16.149 Pa

In order to efficiently control the quality of the Tibetan medicine Gentianae Szechenyii Flos, the quality standard was established in this study. The tests of water content, total ash and ethanol-soluble extractives of the crude drugs were carried out based on the methods recorded in appendix of Chinese Pharmacopeia (2010 edition, volume 1). The TLC method was established by using reference drug and gentiournoside A as reference substance, and a mixture of ethyl acetate-methanol-water-formic acid (7: 1.5: 1: 0.2) as the developing solvent system on silica gel G TLC plate. The content of gentiournoside A was assayed by HPLC on a Ultimate XB-C18 (4.6 mm x 250 mm, 5 μm) column, using methanol-water (0.02% phosphoric acid) (52:48) as the mobile phase at a flow rate of 1.0 mL x min(-1). The column temperature is 25 degrees C and the detection wavelength is at 240 nm. As a result, gentiournoside A and the other constituents were separated and presented the same fluorescence light comparing with the reference substance on TLC detected under the UV light(366 nm). The methodology validation for the assay of gentiournoside A showed that it was in a good linear correlation in the range of 10.01-400.32 mg x L(-1) with the regression equation of Y = 1 539.5X - 33.339 (r = 0.999 7), and the average recovery was 99.68% (RSD 1.92%). The mass fractions of gentiournoside A, water content, ethanol-soluble extractives of 19 batches samples were varied in the ranges of 14.48-31.51 mg x g(-1), 11.25% -12.74% and 24.21% - 31.60%, respectively, and total ash was 4.64% - 6.12% detected from 10 batches samples. The recommended standards of quantitative indexes are that the mass fractions of gentiournoside A and extractives are not less than 15.0 mg x g(-1) (1.5%) and 21.0%, respectively; the water and total ash are not more than 13.0% and 6.0%, respectively.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; standards ; Flowers ; chemistry ; Gentiana ; chemistry ; Medicine, Tibetan Traditional ; Quality Control

The relationship of nutrients and microorganisms in soils with polyphenols and total flavonoids of Houttuynia cordata were investigated by measuring nutrients, enzyme activity, pH, concentrations of microbe phospholipid fatty acids (PLFAs) in soils, and determining concentrations of polyphenols and total flavonoids of H. cordata. The research is aimed to understand characteristics of the planting soils and improve the quality of cultivated H. cordata. The soils at different sample sites varied greatly in nutrients, enzyme activity, pH, microbic PLFAs and polyphenols and all flavonoids. The content of total PLFAs in sample sites was following: bacteria > fungi > actinomyces > nematode. The content of bacteria PLFAs was 37.5%-65.0% at different sample sites. Activities of polyphenol oxidease, concentrations of available P and content of PLFAs of bacteria, actinomyces and total microorganisms in soils were significantly and positively related to the concentrations of polyphenols and total flavonoids of H. cordata, respectively (P < 0.05) . The Content of fungi PLFAs in soils was significantly and negatively related to concentrations of polyphenols and total flavonoids of H. cordata, respectively (P < 0.05). This study provides evidence that effectiveness of the soil nutrient, which may be improved due to transformation of soil microorganisms and enzymes to N and P in the soils, was beneficial to adaptation of H. cordata adapted to different soil conditions, and significantly affects metabolic accumulation of polyphenols and flavonoids of H. cordata.
Bacteria ; metabolism ; Drugs, Chinese Herbal ; analysis ; metabolism ; Fatty Acids ; metabolism ; Flavonoids ; analysis ; metabolism ; Fungi ; metabolism ; Houttuynia ; chemistry ; metabolism ; microbiology ; Phospholipids ; metabolism ; Polyphenols ; analysis ; metabolism ; Soil ; chemistry ; Soil Microbiology

Objective To compare the planning systems of the simplified manual intensity modulated (SMIM) irradiation and standard tangential (ST) irradiation,in order to explore the application of SMIM technique in clinic.Methods In 64 cases of breast cancers after breast conserving surgery,each underwent both SMIM and ST planning systems.SMIM planning was designed by copying additional fields for shielding the high dose areas from internal or lateral tangential field.The high dose areas were reduced by adjusting the size of the additional field and open tangential field.To optimize the SMIM planning,3 high dose areas (＞ 103% ,＞ 105% and ＞ 107%) were shielded and 3 protocols carried out.The wedges were also optimized in ST plan.The target coverage and dose homogeneity and dose of organ at risk were compared between SMIM and ST planning systems.Results When the dose was normalized to cover the volume of 95% CTV,85% of the shielded areas in optimal SMIM planning were that of ＞ 103% high dose area,and 94% of target area was covered.The study on the volume of breast CTV showed that,in the large breast group,SMIM could not only significantly reduce the high dose areas,and the maximum dose as well as the dose of organ at risk,but also enhance the dose homogeneity index.However,no such effect was not significant in the small breast group.Conclusions The simplified manual intensity modulated technique can improve target dose homogeneity in the large breast cases instead of the standard tangential technique.

Objective Multidrug resistance-associated protein 1 (MRP1) has been reported with a close correlation with tumor multi-drug resistance. Real-time quantitative PCR (QRT-PCR) was performed to detect the MRP1 gene expression in childhood acute lymphoblastic leukemia (ALL) and its clinical signiifcance was analyzed. Methods Sixty-seven denovo ALL patients and 10 healthier children as bone marrow donor were studied. The chemotherapy was given according to CCLG-2008 protocol. SPSS software was employed to analyze the data and p-value below 0.05 was regarded as statistic signiifcance. Results MRP1 expression level showed a close correlation with ALL risk, the median of MRP1 expression was 4.28 (2.75~6.12), 5.62 (4.99~8.60) and 7.56 (3.66~11.13) for standard-risk group (SR), intermediate-risk group (IR) and high-risk group (HR), respectively. MRP1 mRNA expression in T-ALL group was 7.71 (6.49~14.35), which is higher than that of B-ALL (5.18(3.89~8.46)) (P<0.01). The rate of leukemia cells’ sensitivity to prednisone on 7th day was 70.6%in high expression group (n=34), which was signiifcantly lower than that in low expression group (n=33, 90.9%, P=0.035). The complete remission rateon 33th day was 64.7%in high expression group, and 87.9%in low expression group, which showed a signiifcant difference between them (P=0.026). Conclusions In children ALL, the expression of MRP1 is closely related with immunophenotyping, treatment response, hazard level and disease relapse.

Objective: To analyze and review the clinical efficacy of acupuncture (including electroacupuncture) alone for allergic rhinitis (AR) and to compare its efficacy with antihistamines and Chinese patent medicineBi Yan Kang Tablet. Methods: The search strategy, inclusion and exclusion criteria were made according to the principle of evidence-based medicine. We performed a systematic search on China National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP), Chinese Biomedical Literature Database (CBM), PubMed, Excerpta Medica Database (EMBASE), Web of Science, Cochrane Library, and Cochrane Central Register of Controlled Trials (CENTRAL) for randomized controlled trials (RCTs) of acupuncture for allergic rhinitis between January 1990 and December 2015. The quality was evaluated by Cochrane Handbook for Systematic Reviews of Interventions Version 5.1, and the meta-analysis was conducted by RevMan 5.3 version. Results: Twenty eligible RCTs were included into the meta-analysis after selection. Compared with antihistamines, the meta-analysis showed RR=1.24>1, 95%CI[1.15, 1.33],P<0.00001, indicating that acupuncture achieved a better total effective rate for AR than antihistamines; MD=–0.93<0, 95%CI[–1.22, –0.63],P<0.00001, indicating that acupuncture is better than antihistamines in decreasing the total nasal symptom score (TNSS) in AR patients; and MD=1.46>0, 95%CI[–10.84, 13.75],P=0.82, indicating that there was no statistical difference between acupuncture and antihistamines in regulating immunoglobulin E (IgE) in AR patients. Compared withBi Yan Kang Tablet, the meta-analysis has shown RR=1.50>1, 95%CI[1.30, 1.73],P<0.00001, indicating that acupuncture achieved a better total effective rate for AR than Chinese patent medicineBi Yan KangTablet. Conclusion: Acupuncture alone can achieve a better total effective rate for AR than antihistamines andBi Yan Kang Tablet. It is also better than antihistamines in improving clinical symptom scores; however, whether acupuncture is better thanBi Yan KangTablet needs further proof. As far as current data are concerned, there was no statistical difference between acupuncture and antihistamines in improving serum IgE; further study is needed in this regard. The risk of bias due to absent randomization methods or blinding implementation decreased the evidence level of the overall conclusion.

Objective The purpose of this study was to prepare RGD4C modified ferritin nanocages (RGD4C-FRT) for targeted drug delivery to rat hepatic stellate cells (HSC-T6). Methods RGD4C modified human H-chain ferritin was expressed and purified. Doxorubicin (Dox) was encapsulated into the cavity of RGD4C-FRT through ion channels, which resulted in RGD4C-FRT-Dox. The target of RGD4C-FRT-Dox to HSC-T6 was detected using fluorescence microscopy. Results Transmission electron microscopy showed that RGD4C-FRT was hollow spherical-structured with uniform size and good dispersion. The average particle diameters of RGD4C-FRT and RGD4C-FRT-Dox were 12.57 nm and 20.13 nm , respectively. The drug encapsulation efficiency and loading percentage of RGD4C-FRT-Dox were 77.32% and 15.88% respectively. RGD4C-FRT-Dox was significantly uptaken by HSC-T6, and the uptake could be blocked by the empty carrier RGD4C-FRT. Conclusion RGD4C-FRT-Dox can specifically target HSC-T6. Further animal experiments are needed to inspect its therapeutic effect on liver fibrosis.

Objective: To investigate the role of human umbilical vein endothelial cells (HUVEC) in supporting hematopoiesis and the expansion of hematopoietic stem/progenitor cells in vitro. Methods: According to the fact that HUVEC supernatant has colony stimulating activity shown by methylcellulose colony-forming assay and HUVEC can maintain the survival of mononuclear cells for at least four weeks in vitro, CD34+ cells from umbilical cord blood were seeded with (HUVEC group) or without (control group) HUVEC monolayer. Every week cells were collected and counted, the frequency of CFU-GM was measured by using methylcellulose colony-forming assay, and the percentage of CD34+ and CD41a+ cells was measured by flow cytometry. Results: In control group，all the CD34+ cells died in two weeks. However, in HUVEC group,most nucleated cells and CD34+ cells were expanded by 68.1±14.8 fold and 6.6±1.4 fold，respectively at the third week while CFU-GM expansion reached its peak (5.7±2.1 fold) at the week 2. Moreover, the percentage of CD41a+ cells was enhanced significantly, reaching a maximum (15.6%) at the week 3. Conclusions:HUVEC can support hematopoiesis in vitro and expand the hematopoietic progenitor cells and CD41a+ cells in direct contact coculture.