From the consideration of the requirement of the state traditional Chinese medicine ( TCM ) clinical research facility (Hubei), this article focused on the establishment of the study and exploitation of TCM plat-form which serves exploitation of the hospital pharmaceuticals and new pharmaceuticals of key diseases such as liver disease . After 5 years of construction , the platform with innovative ability was initially formed; some characteristic pharmaceuticals were exploited, which promoted the clinical service ability of key disease, brought good economic and social influence . This article made a preliminary discussion on the construction achieve-ments , experiences and expectations of this platform .

Objective To analyze the clincopathological features and prognosis of 77 colorectal cancer patients with synchronous liver metastases after resection of the primary tumors.Methods Seventy seven colorec-tal cancer patients with synchronous liver metastases who underwent resection of the primary tumors and patholog-ical confirmed were enrolled in this retrospective study.The clincopathological features and prognosis were ana-lyzed.Kaplan-Meier method was used to analyze survival rate.The differences between survival curves were cal-culated by Log-rank test.Results The analysis showed that sex,tumor differentiation and TNM stage were as-sociated with prognosis(P<0.05).The analyses demonstrated no differences in age,primary tumor site,treat-ment,number of liver metastases and extrahepatic metastasis(P>0.05).There was a significant correlation be-tween CEA,CA19-9 level and gender,tumor differentiation and TNM stage(P<0.001)and the corresponding surgical procedure and TNM stage(P<0.05).Conclusion The analysis show that male,the low differentiation and stageⅢare poor prognosis factors.

Objective To construct the recombinant plasmid containing human filaggrin gene,purify and identify the immunoreactivity of the recombinant protein,and establish the indirect ELISA to detect AFA for diagnosis of RA.Methods The constructed plasmids were transformed into E. Coli Rosettagami(DE3).This fusion protein was purified by NAT chromatography.ELISA coated with the fusion protein Was established to detect the AFA in serum of patients,which included 114 cases of RA,56 cases of SLE,32 cases of OA and 40 cases of normal controls. The correlation between the results of AFA and anti-CCP in RA group were compared. Results 321 bp fragment of filaggrin gene was amplified and the recombinant expression vector pET-28a( + )-filaggrin was constructed. The sequence of filaggrin gene was the same as the sequence reported in the literatures. The Rosetta-gami (DE3) strains of E. Coli with recombinant vector showed high level of filaggrin protein after induction. The SDS-PAGE showed that the plasmid expressed the filaggrin fusion protein with molecule weight of 14 000 Da. The expression protein could be purified by Ni-NAT with activity. The absorbance value of AFA in RA group was 0.473 ±0. 248 while they were 0. 160 0. 088, 0. 121±0. 070, 0.050 0. ±018 in SLE, OA and normal groups respectively. There were significant differences of absorbance values of AFA between RA and SLE, OA, control group (t = 12.004, 14. 464, 18.078, P<0. 01, respectively). The positivities of anti-filaggrin in RA, SLE and OA were 48.2%, 5.4% and 3. 1% respectively. The positivities of AFA were significantly different between RA, OA and normal control groups (x~2 = 67. 088, P < 0. 01). There was positive correlation of results between AFA and anti-CCP antibody (r = 0.42, P < 0. 05 ) . The consistency rate of results between AFA and anti-CCP was 70. 1%. Anti-CCP was negative in 10 out of 114 patients with AFA positive. AFA can be used to diagnose RA with sensitivity of 48. 2% , specificity of 96.9% , positive predictive value of 93. 2% and negative predictive values of 67. 9% . Conclusions The purified human filaggrin fusion protein is successfully purified. The indirect ELISA method based on the recombinant protein shows good sensitivity and specificity. Joint detection with AFA and anti-CCP can improve the positive rate of detection.

Objective To observe the different needs of Uighur and Han stroke patients for oceupational activities using the Canadian occupational performance measure (COPM).Methods The COPM was employed to evaluate 51 stroke patients in hospital before and after treatment.Thirty were from the Han ethnic group and 21 were Uighur.The first evaluation was performed at admission to confirm their occupational activity problems.Interventions were then planned using a patient-centered occupational therapy model.At discharge the second evaluation was performed to assess and compare the effects of treatment.Results In the first evaluation,both the Uighur and Han stroke patients had problems with self-care activities which were more prominent than those with productive and leisure activities.Older Uighur patients had special occupational activity needs resulltng from their religious practices.Compared witb the first evaluation,the total performance and satisfaction scores had improved significantly by the second evaluation for both the Han and Uighur patients,but their satisfaction scores were lower than their occupational activities performance scores.Conclusion Different nationalities may have different occupational activities needs.The COPM is easy to use and helpful in confirming occupational activity problems.Its use can contribute to the plan ning of primary goals for rehabilitation and treatment programs and help assess the effect of rebabilitation.

Objective To explore the expression of Metastasis-associated in colon cancer-1 (MACC1) in patients with nasopharyngeal carcinoma (NPC), and its relationship with clinical pathologic characteristics and prognosis Methods The expression of MACC1 was detected in 130 cases of NPC and the relationship among the MACC1 expressions, clinical pathologic characteristics and prognosis of NPC was analyzed. Results Positive expression rate of MACC1was 68.5% in the NPC and MACC1 expression was associated with advanced T stages, lymph node metastasis and advanced clinical stages of NPC (P < 0.05). The results of Kaplan-Meier survival analysis showed that the five year overall survival rate in patients with positive expressions of MACC1 (45.9%) was significantly lower than that of those with negative expressions (73.7%) and the difference was statistically significant (P < 0.05), Cox multi-factor analysis results showed that MACC1 expression was an independent prognostic factor for NPC (P = 0.004). Conclusion MACC1 abnormal expression is closely related to the invasion and metastasis of NPC and it is expected to become a new target for gene therapy of NPC.

To establish the quality standard for Zhike Xiaoyan Zhitong lotion. Methods: Phellodendri chinensis Cor-tex, Angelicae sinensis Radix and Astragali Radix in the preparation were identified by TLC. The content of berberine hydrochloride, the effective component in Phellodendri chinensis Cortex, was determined by HPLC with the chromatographic conditions as follows: an Agilent 5 TC-C18 column (250 mm × 4. 6 mm, 5 μm) was used, the mobile phase was acetonitrile-0. 05 mol·L-1 potassium dihydro-gen phosphate (30∶70) with the flow rate of 1. 0 ml·min-1 , the detection wavelength was 265 nm, and the column temperature was at 30 ℃. Results:The spots of Phellodendri chinensis Cortex, Angelicae sinensis Radix and Astragali Radix in Zhike Xiaoyan Zhitong lotion on TLC were clear with strong specificity and without any interference from the negative controls. There was a good linear rela-tionship for berberine hydrochloride within the range of 0. 029-3. 628 μg(r=0. 999 9), and the average recovery was 97. 83% (RSD=2. 05%, n=6). Conclusion:The qualitative and quantitative methods are accurate, reliable and repeatable, which can effectively control the inherent quality of Zhike Xiaoyan Zhitong lotion.

Objective To establish an HPLC method for the content determination of protodioscin and diosgenin in Dioscoreae Hypoglaucae Rhizoma from different habitats.Methods The chromatographic separation was performed on an Agilent C18 column (4.6 mm×150 mm, 5μm) with mobile phase of acetonitrile-water solution with gradient elution at the flow rate of 0.8 mL/min; the detection wavelength was 208 nm; the column temperature was 30℃; the injection volume was 20μL.Results Protodioscin showed a good linear relationship among the range of 1.73–8.64 μg (r=0.999 6), with the average recovery of 101.98% (RSD=1.53%); Diosgenin showed a good linear relationship among the range of 1.03–8.20μg (r=0.999 1), with the average recovery of 101.60% (RSD=2.41%). The contents of protodioscin and diosgenin in Dioscoreae Hypoglaucae Rhizoma from 10 different habitats were in the range of 0.89%–2.24% and 0.75%–3.22%, respectively.Conclusion The method is simple, accurate and with repeatability, which can be used as quality control method of Dioscoreae Hypoglaucae Rhizoma.

Crystallography, X-Ray ; Hepatitis A ; genetics ; virology ; Hepatitis A virus ; chemistry ; pathogenicity ; Humans

Aim To investigate the effect of gelsemium alkaloids on chloride channels and cell volume in he-patic carcinoma cells. Methods The time-lapse live cell imaging and whole-cell patch clamp techniques were used respectively to detect the volume changes and currents induced by gelsemium alkaloids in HepG2 cells. Results It was found that the cell volume was decreased by (12. 48 ± 2. 2) % (P<0. 01) when ex-posed to gelsemium alkaloids for 50 min and this phe-nomenon could be inhibited by the chloride channel blocker tamoxifen. It was shown by whole-cell patch clamping that a chloride current could be evoked by extracellular application of gelsemium alkaloids ( 2μmol·L-1 ) . The current was outward-rectified with-out obvious voltage- and time-dependent inactivation. The reversal potential of the current was ( -3. 21 ± 0. 67) mV ,which was close to the equilibrium poten-tial of chloride. The extracellular application of the chloride blockers, tamoxifen and 5-notro-2-(3-phenyl-propylamino)benzoic acid (NPPB), and 47% hyper-tonic solution inhibited the current significantly ( P <0. 01 ) . Conclusion Gelsemium alkaloids could acti-vate chloride channels and induce a volume decrease ( named apoptotic volume decrease, AVD) , and these effect could be inhibited by chloride channel blockers. The results suggest that the chloride channel can be one of the targets of gelsemium alkaloids in their anti-cancer action.

Objective To study the effect of mitogen-activated protein kinas/extracellular signalregulated kinase (MAPK/ERK) signaling pathway on cell proliferation modulated by Sonic Hedgehog (Shh) signaling in fibroblast-like synoviocytes (FLS) isolated from patients with active rheumatoid arthritis (RA).Methods The synovial tissue were collected by the synovial arthroscopic debridement or arthroscopic synovectomy of RA patients with active disease activity [disease activity score(DAS)28 ≥3.2].The RA-FLS were primarily cultured by the explanted culture,and then were treated with Shh agonist purmorphamine,inhibitor cyclopamine or MAPK/ERK signaling pathway inhibitor U0126,respectively.Western blotting was used to examine the phosphorylation level of ERK 1/2 (p-ERK1/2),which was the critical protein of MAPK/ERK signaling.The cell proliferation activity was detected using cell proliferation and cytotoxicity kit-8 (CCK8),and the cell proliferation rate was detected using a flow cytometry.Analysis of variance and Kruskal-Wallis H(K) test were used for statistical analysis.Results Compared with the control group,purmorphamine transiently increased p-ERK1/2 protein at the concentration of 1 μmol/L,and the peak activations of p-ERK1/2 took place at 15 min (P＜0.01).Cyclopamine and U0126 decreased the expression ofp-ERK1/2 protein (P＜0.01).After the RA-FLS treated with purmorphmine(1 μmol/L)for 48 hours,the cell proliferation activity was (114±4)％ and the percentage of S phase cells was (8.39±0.60)％,which was significantly higher than those of the control group (100±0)％ (P＜0.01) and (3.29±0.69)％ (P＜0.01).After treated with cyclopamine (10 μmol/L) for 48 hours,the cell proliferation activity of RA-FLS was (89±1)％ (P＜0.05) and the percentage of S phase cells was (1.53±0.22)％ (P＜0.05).When co-treated with purmorphamine (1 μmol/L) and U0126 (10 μmol/L),the cell proliferative activity was (89±2)％ (P＜0.05) and the percentage of S phase cells was(1.07±0.25)％(P＜ 0.05).Conclusion Shh may promote proliferation of RA-FLS via modulating MAPK/ERK signaling,which in turn contributes to hyperplasia of synovium and ultimately leading to RA.