AIM: To discuss the application of overlapping ratio to the fingerprints of plant and plant extract. METHODS: We took the HPLC fingerprints of green tea and tea polyphenols for decision procedure. RESULTS: Overlapping ratio could be applied to selecting representative samples for characteristic fingerprints of green tea and tea polyphenols,and could be used to deduce the raw material,process and producer of tea polyphenols.(CONCLUSION:) Overlapping ratio can be used to choose the suitable samples for establishing the characteristic fingerprint spectrum,and can supply references to relationship investigation among products,raw material and process.

Objective: To develop a HPLC method for simultaneous determination of catechins (EGC、DC、EGCG、EC、GCG、ECG) and caffeine in tea polyphenols and to quantify seven ingredients in twenty nine batches of tea polyphenols samples from thirteen producers. Methods: The study was achieved using a C 18 column with a methanol water 0.1% formic acid gradient elution system. Results: The peak resolutions of seven ingredients in mixed standards were all above 2; The resolutions in samples were all above 1.5 except EGCG; The average recoveries were 98.56%～100.14%. Conclusion: This method is accurate, stable, reproducible and suitable for quality control of tea polyphenols; the variations of contents are obvious among tea polyphenols samples from different even the same producers.

To explore the clinical characteristics and imaging features of miliary tuberculomas in central nervous system (CNS).A total of 5 cases diagnosed with tuberculosis in CNS first diagnosed by neurologists in Navy General Hospital of PLA were enrolled in the study.All clinical and imaging data were collected and analyzed retrospectively.The main initial symptoms were fever and headache (4/5).Multiple diffused miliary lesions were shown by brain MRI,with maximum diameter ranged from 1-4 mm and ringshape or nodular enhancement after gadolinium injection.As mycobacterium tuberculosis could seldomly be found in serum and cerebrospinal fluid,contrast MRI remains the effective method for detecting miliary tuberculomas in CNS.

Objective To analyze the clinical,imaging and pathological features of cortical vein thrombosis (CoVT).Methods Three cases of cortical vein thrombosis were diagnosed in our hospital from February 2010 to October 2012.We reviewed and summarized their clinical manifestation,radiological feature and pathological characteristics.Results All patients were young with acute onset.The main clinical manifestations included headache,epilepsy or limbs weakness.Two cases had intracranial hypertension.One case had decreased activity of protein S.One had increased plasma homocysteine.Brain computed tomography scan showed hypodensity lesion with some hyperdensity inside.Cranial routine magnetic resonance imaging showed long signal in T1-weighed image and T2-weighed image,with occasional short T1 signal inside.Contrast-enhanced magnetic resonance imaging revealed heterogeneous enhancement.All of 3 cases underwent brain biopsy because of the suspected diagnosis of brain tumor.Brain pathology showed the local necrosis and hemorrhage,dilated small vein with congestion or thrombosis.Neuronal degeneration,hyperplasia of gliocyte,hyperplasia of endotheliocyte in small blood vessels with reaction of histiocytes was also displayed.Duration from initial visit to final diagnosis was from 14 days to 2 months.Conclusions CoVT has various clinical and radiological manifestations and it is easy to misdiagnose as brain tumor.Careful analysis of clinical and imaging data could improve its diagnostic accuracy.Brain biopsy would also be helpful for diagnosis.

Objective To explore the effect of endovenous laser treatment ( EVLT) for varicose great saphe-nous veins complicated with ulcers .Methods 40 cases ( 46 limbs ) of varicose great saphenous veins complicated with ulcers underwent EVLT .The abnormal vein branches of the base of ulcers and adjacent ulcers were burned and closed.Results Except a case suffered the secondary transplantation for skin flap necrosis ,no infection,skin necro-sis,deep vein thrombosis complications occurred in other 39 cases.All the ulcers healed ,and postoperative lower limb varicose veins were disappeared .The operation time was (65 ±11.6) min.All the ulcers healed and the hospitalization time was (21 ±4.6)d.All the patients were followed up for 6-36 months,and no recurrence was found.Conclusion EVLT for varicos e great saphenous veins complicated with ulcers has the advantages of good effect ,minimal incision , and quick recovery .

Objective To study the clinical significance of intercostobrachial nerve ( ICBN) reservation in radical resection of breast cancer .Methods 42 cases with breast cancer at stageⅠ,ⅡorⅢa were randomly divided into experimental group(n=28) and control group(n=14).In the experimental group,the ICBN were preserved in the axillary lymph node dissection operation ,while in the control group ,the ICBN were resected in the operation .After the operation,both two groups were treated as breast cancer and the patients were followed up .The operation time,the number of ALND , local tumor recurrence and distant metastasis rates after operation , survival rate were compared between the two groups.Results Postoperative 14 days,3 months and 6 months,the incidence rates of the sensory disorder in the internal upper arms in the control group were 85.7%,64.3% and 50.0% respectively,those in the experimental group were 10.7%,3.5% and 0.0%.The differences were statistically significant between the two groups(P<0.05).The average operation time,average number of lymph node dissection between control group and experimental group had no significant difference [(80.0 ±12.9) min vs (88.0 ±15.6) min,(16.4 ±5.4) vs (14.8 ±6.1)](P >0.05).In the control group,the local recurrence rate,distant metastasis rate and survival rate were 0.0%,7.0%and 0.0%,those in the experimental group were 0.0%,0.0% and 0.0%.There were no statistically significant differences between the two groups .Conclusion It is feasible to preserve the ICBN in radical resection of breast cancer ,which reserves the skin sensory function of the internal upper arm and axilla ,and enhance the patients'quality of life.

Objective:To express mycobacterium tuberculosis protein MPB 64 in baculovirus insect cell expression system ,and identify its immunogenicity.Methods:The target gene MPB64 was connected to pFastBac vector ,then the pFastBac-MPB64 plasmid which was harvested would transformed to DH10Bac competent,and the target gene was transposition into Bacmid by Tn7 transposase fragment,therefore Bacmid-MPB64 Shuttle vector was obtained.The shuttle vector was packaged by liposomes and transfected Sf 9 cells to harvest P1-generation virus ,then high titers of P4 generation virus was harvested by repeat transfected Sf 9 cells three times.The target protein MPB64 was purified from the supernatant by Q Sepharose FF and Ni affine chromatography ,which were used to immunize BALB/c mice.Antibody changes in serum would be detected ,and the proliferation of immunized mice spleen cells would be detected by MTT,detected the IFN-γsecretion by MPB64 stimulated spleen cells by ELISA method.Results: MPB64 successfully expressed in insect cells.The purity of target protein was over 90% and yield up to 35 mg/L after purification.Purified protein can effectively stimulate BALB/c mice to produce antibodies , increase the content of IFN-γmedium in mice spleen cells ,and significantly promoting proliferation in spleen cells between 0.2-100 μg/ml.Conclusion: MPB64 which has immunogenicity was successfully expressed in baculovirus insect cell expression system ,that open a new avenue for tuberculosis vaccine production.

Objective To explore the fibrosis mechanism of patients with fibrosing cholestatic hepatitis(FCH) in the way of degradation of collagen.Methods The expressions of matrix metalloproteinase 1(MMP1) and tissue inhibitor of metalloproteinase 1(TIMP1),and contents of type I,III collagen proteins were detected by immunohistochemical staining in the liver tissues of 9 cases with FCH associated with HBV developed following renal transplantation and 5 cases without liver disease as controls.Results The expressions of MMP1 and TIMP1,and type I,III collagen proteins in the patients with FCH were significantly higher than those in the control group.There was a positive correlation between the expressions of type I,III collagen proteins and TIMP1/ MMP1 ratio.Conclusion Hepatic fibrosis in the patients with FCH associated with HBV developed following renal transplantation may be relative to the increase of TIMP1 expression which inhibit the degradation of collagen.

Objective To observe the expressions of matrix metalloproteinase 1(MMP1) and tissue inhibitor of matrix metalloproteinase1(TIMP1) and the collagen type Ⅰ,Ⅲ deposition in the liver tissues,and evaluate the possible fibrosis mechanism of patients with chronic hepatitis B(CHB) in the way of degradation of collagen. Methods The specimens of the biopsy liver in 50 cases with CHB were detected for the expressions of type Ⅰ,Ⅲ collagen proteins,MMP1 and TIMP1 by immunohistochemical staining. Results The expressions of type Ⅰ,Ⅲ collagen proteins and TIMP1,were significantly increased along with the advancing of hepatic fibrosis.There was a positive correlation between the expressions of type Ⅰ,Ⅲ collagen proteins and TIMP1. Conclusions Hepatic fibrosis in the patients with CHB may be related to increase of TIMP1 expression that inhibit the degradation of collagen.

BACKGROUND:Pueraria is a member of isoflavone characterizing by decreasing the toxic effects of alcohol,relieving the effect of alcohol withdrawal,anti-oxidative effect,removing oxygen free radicals,and preventing cell injury induced by lipid peroxidation.OBJECTIVE:To investigate the inhibitory effect of puerarin on adipogenic differentiation of alcohol-induced human bone marrow mesenchymal stem cells (hBMSCs).DESIGN,TIME AND SETTING:An in vitro observation based on cytology was performed at Laboratory of Biochemistry and Molecular Biology,Basic Medical College of Zhengzhou University in October 2008.MATERIALS:The bone marrow was taken from the adult healthy volunteers from the Department of Orthopedics of the First Affiliated Hospital of Zhengzhou University.Puerarin standard was provided by the National Institute for the Control of Pharmaceutical and Biological Products.METHODS:The primary hBMSCs were purified from the adult marrow by gradient centrifugation.The second passaged cells were divided into 3 groups.Blank control group:Cells were cultured with DMEM culture media containing 10% fetal bovine serum;Model group:Cells were cultured with 0.09 mol/L alcohol;Experimental group:Cells were cultured with 0.09 mol/L alcohol and 10-6 mol/L puerarin;while,the culture liquid was changed every two or three days.Cells were cultured for 7 days in each group.MAIN OUTCOME MEASURES:PPARγ 2 gene expression with RT-PCR;amount of adipocyte with oil red "O" staining;alkaline phosphatase activity.RESULTS:PPAR-γ2 mRNA expression in the model group was significantly greater than experimental group and blank control group (P ＜0.05),while the expression in the experimental group was slightly greater than blank control group,but there was no significant difference (P ＞0.05).The number of adipocytes was the most in the model group,and the lipid droplet was large and abundant.The number of adipocytes in the experimental group was significantly less than model group (P ＜ 0.05),and there were few adipocytes in the blank control group.Compared with blank control group,alkaline phosphatase activity was decreased in the model group (P＜0.05);but the activity in the experimental group was increased compared to model group (P ＜ 0.05).CONCLUSION:Puerarin inhibits differentiation of hBMSCs into adipocytes induced by alcohol.