Composition of invention patent application is important for obtaining patent right and maintaining the right stability. According to the features of TCM invention patent application and common problems in examination, this article summarized several key points for effective application composition, including composing background technology objectively, highlighting technical problems, reasonable layout of technical pian, working in contact with technical effects, and clarifying technical terms. It expounded each point with relevant typical cases, with a purpose to provide references for applicants and agents of TCM invention patent application.

Objective:To study the effect of miR-122a on inhibition proliferation of laryngeal carcinoma cell line Hep-2.Meth-ods:The oligomucleotide of miR-122a was transfected into laryngeal carcinoma cell line Hep 2 cells,which were devided into three groups of A(miR-122a transfection),B(miR-122a inhibitor),C(miR-122a-NC inhibitor) and group of D(blank control).The expres-sion of miR-122a was defected by RT-PCR,and relevant protein expression was evaluated by Western blot .The cell proliferation and cell cycle were determined by MTT assy and flow cytometry ,respectively.Results:Compared to group D,miR-122a expression in Hep2 cells was obviously elevated atter miR-122a-transfected.The proliferation of Hep2 cells in group A was significantly inhibited and the cell cycle arrested at G1/G0 phase.The protein expression of CDC42 was downregulated with decreased expressions of CDK 4 and cyclin D1 in group A.Conclusion:miR-122a inhibits the proliferation activity of Hep 2 cells,suggesting that miR-122a can be taken as a po-tential candidate for gene therapy of laryngeal carcinoma .

Objective To develop a HPLC quantitatively test method of glycyrrhizin, liquiritin and isoliquritigenin simultaneously for licorice. Methods The 67% methanol extracts of licorice powder were analyzed by reverse-phase HPLC on an ODS column, using a gradient elution with a mixture of 1% H3PO4 and acetonitrile. The eluate was monitored by UV absorption at 248, 276, 360 and 370 nm. Results The linear equation of glycyrrhizin was Y =1?106X +16 220, r 2=1. The linear equation of liquiritin was Y= 2?106X+49 444, r 2=0.999 5. The linear equation of isoliquritigenin was Y =1?107X +4.466 7, r 2=1. The recovery rate of glycyrrhizin, liquiritin and isoliquritigenin was 98.01%, 102.63% and 98.18% respectively. Conclusion The method is accurate, stable amd reliable, and can be used to determine the three main chemical contents simultaneously.

Objective To study the protective effect of Duzhong Hongjingtian Capsule (DHC) on cerebral ischemia in rats. Methods Thrombotic ischemia and middle cerebral artery occlusion (MCAO) induced by FeCl3 in rats was used to observe the protective effect of DHC on ischemia stroke, and common carotid artery occlusion (CCAO) rats was used to analyze the effect of DHC on blood brain barrier (BBB) permeability. Results DHC could improve the abnormal behavior of MCAO rats and decrease the infarction area and water content in the brain of MCAO rats. DHC could also improve the cerebral blood flow on MCAO rats and significantly reduce the permeability for evans' blue (EB) of CCAO rats. Conclusions DHC can protect the brain ischemic injury.

Objective To investigate the expression of CD40/CD40L on peripheral blood mononuclear cells (PBMC) in patients with recurrent genital herpes (RGH). Methods Flow cytometric analysis was employed to identify the expression of CD40 and CD40L on PBMC and the T lymphocyte subgroups in the peripheral blood of 30 patients with RGH and 20 healthy controls. Results Percentage of the cells with either CD40 or CD40L expression in the patients with RGH was significantly lower than that in the controls (P = 0.0061 and 0.041, respectively). The RGH group had a significantly lower percentages of CD3+ T cells (P=0.025) and CD4+ T cells (P = 0.032) as compared to the controls. Conclusion Reduced costimulatory interaction of CD40 and CD40L in RGH patients may be one of the important factors for the recurrence.

To compare the effects of polysaccharide nucleic acid fraction of bacillus calmette guerin (BCG-PSN) and thymopeptides on T-lymphocytes of normal and immunosuppressed mice, CD4+ and CD8+ T-lymphocyte subsets of single nucleic cell in thymus, spleen and peripheral blood were detected successively by flow cytometry after application of BCG-PSN and thymopeptides. Meanwhile, CD4+/CD8+ ratio was also calculated. The results showed that both BCG-PSN and thymopeptides could decrease the proportion of CD4+ CD8+ T-lymphocyte subsets in the thymus, at the same time increase CD4+ T-lymphocyte, CD8+ T-lymphocyte proportion in the three tissues. The fluctuation in amplitude was greater in thymopeptides group than that in BCG-PSN group. It is concluded that acting location of thymopeptides is in thymus, its stimulating action is stronger than that of BCG-PSN, while BCG-PSN not only accelerates the differentiation in thymus, but also has some direct stimulation to peripheral CD4+ T-lymphocytes, and can maintain CD4+/CD8+ ratio within normal range. So, BCG-PSN is safer.
Adjuvants, Immunologic/*pharmacology ; Immunocompromised Host ; Mycobacterium bovis/*chemistry ; Nucleic Acids/pharmacology ; Peptide Fragments/*pharmacology ; Polysaccharides, Bacterial/*pharmacology ; T-Lymphocyte Subsets/*drug effects ; Thymus Gland/chemistry

Objective To analyze the patients with infection before transfusion in onder to avoid misdiagnosis and blood transfusion caused by cross-infection and remind medical staff attention to operation and self-protection.Methods HBsAg,Anti-HCV,Anti-TP,Anti-HIV1/2 were detecced by 2 576 cases of pre-operative,prenatal,blood transfusions before the patient's blood ELISA assay.Results 2 576 cases of infectious markers in patients with serum positive rate was 12.15％.The positive rates of was HBsAg,Anti-HCV,Anti-TP,Anti-HIV1/2:11.06％,2.13％,1.67％,0.00％,HBsAg and Anti-HCV were positive for both the percentage of 0.54％.Conclusion The transfusion of pre-infection detection is necessary for patients with early diagnosis and treatment of infectious diseases to avoid and prevent hospital infections,medical personnel,as well as occupational infection to reduce or avoid blood transfusion caused by medical disputes.

Objective To investigate the infection of Xuebijing injection on inflammatory factor of large sized avulsion patients.Methods 70 patients being selected with large sized avulsion were randomly recruited into a treatment group(35 patients)and a control group(35 patients).The control group received traditional comprehensive treatment.On this basis,Xucbijing injection was injected to the treatment group on admission day,and Xuebijing injection 50 ml in 0.9%NaCl solution 100 ml was,intravenously infused for 60 min once,2 times daily,up to 7 d.TNF-α,IL-6,CRP,WBC and NEU%of the two groups were respectively detected before treatment and 7 days after the treatment.Exudation of wound surface was also observed.Results After 7 days of treatment,there was significant difierence in the treatment group compared with pretreatment(P＜0.01).There was statistical difference between two groups after 7 days of treatment(P＜0.01).Exudation of wound surface of the treatment group was less than the control group's.Conclusion Xuebijing injection has antagonistic effect on inflammatory factor of large sized avulsion patients and can lessen exudation of wound surface.

AIM:To prepare Wurenchun Dispersible Tablet(Fructus schisandrae Chinensis)and investigate the dissolution of it.METHODS:The optimal formulation of Wurenchun Dispersible Tablets was established by the single factor experiments.The dissolution of deoxyschisandrin from Wurenchun Dispersible Tablets was determined by HPLC.RESULTS:Wurenchun Dispersible Tablets prepared by the mixture of 50 g extraction and 125 g CaHPO_4 plus excipent(MCC∶CMS-Na 3∶1)could disintegrate and disperse well within 3 min in(20?1)℃ water.The dissolution of deoxyschisandrin from Wurenchun Dispersible Tablets was higher than 80% in 15 min.CONCLUSION:Wurenchun Dispersible Tablets were stable,dissolved fast and completely.The preparation could promote the dissolution of total lignans.

Objective The mutation of the ABCB6 gene is involved in a variety of diseases , including dyschromatosis univer-salis hereditaria (DUH).This study aimed to construct the expression vectors for the ABCB 6-DsRed fusion proteins, pDsRed-wt-AB-CB6 and pDsRed-L356P-ABCB6, detect its cellular localization in A375 cells, and thus facilitate future studies on the pathogenesis of ABCB6-related diseases . Methods The recombinant plasmids pDsRed-wt/L356 P-ABCB6 were constructed based on the previously constructed pIRES2-ZsGreen1-ABCB6 vector and then transfected into A 375 cells.At 48 hours after transfection , the expression of AB-CB6 was detected by Western blot and the cellular localization of ABCB 6 determined under the laser scanning confocal microscope . Results The expression vectors pDsRed-wt/L356P-ABCB6 were verified by colony PCR, enzyme digestion, and DNA sequencing. The expression of ABCB 6 was significantly increased in the A 375 cells after transfected with the recombinant plasmids .Confocal mi-croscopy showed the localization of both wild-type and mutant ABCB6 in the cytoplasm. Conclusion The expression vectors for wild-type and mutant ABCB6-DsRed fusion protein were successfully constructed and the localization of ABCB 6 in A375 cells was de-termined, which may serve as a basis for further studies of ABCB 6 and the pathogenesis of ABCB 6-related diseases .