With the application of monoclonal antibody technology more and more widely, its production technology is becoming more and more perfect. Small molecule monoclonal antibody technology is becoming a hot research topic for people. The application of traditional Chinese medicine small molecule monoclonal antibody technology has been more and more widely, the technology for effective Chinese medicine component knockout provide strong technical support. The preparation of monoclonal antibodies and small molecule knockout technology are reviewed in this paper. The preparation of several steps, such as: in the process of preparation of antigen, hapten carrier coupling, coupling ratio determination and identification of artificial antigen and establishment of animal immunization and hybridoma cell lines of monoclonal antibody, the large-scale preparation; small molecule monoclonal antibody on Immune in affinity chromatography column method is discussed in detail. The author believes that this technology will make the traditional Chinese medicine research on a higher level, and improve the level of internationalization of Chinese medicine research.
Animals ; Antibodies, Monoclonal ; chemistry ; genetics ; immunology ; Humans ; Hybridomas ; metabolism ; Immunologic Techniques ; methods ; trends

Selection and affinities of Cd binding peptides were assayed by phage random dodecapeptide library and affinity chromatography. Two Cd binding peptides were obtained, it was found that the affinities of Cu~ 2+ ,Co~ 2+ ,Zn~ 2+ ,Ni~ 2+ for Cd binding peptides were higher than that of Cd~ 2+ and Cr~ 2+ after detection of the amplified Cd binding peptides displayed phages to different heavy metal-charged resins; the detoxification of E.coli to Ni~ 2+ and Cd~ 2+ was enhanced when infected by Cd binding peptide displayed phages as compared with those of the control. The interaction of Cd binding peptide displayed phages with heavy metals charged resins was also observed under microscope. The work would be of great value and consequences for the study of interaction between heavy metals and proteins(peptides), as well as thedetoxification and bioremediation of heavy metals.

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

Glomerular hypertrophy is the main pathological characteristic in the early stage of diabetic nephropathy (DN), and its regulatory mechanism is closely related to mammalian target of rapamycin (mTOR) signaling pathway activity. mTOR includes mTOR complex 1 (mTORC1) and mTOR complex 2(mTORC2), in which, the upstream pathway of mTORC1 is phosphatidylinositol-3-kinase (PI3K)/serine-threonine kinase(Akt)/adenosine monophosphate activated protein kinase(AMPK), and the representative signaling molecules in the downstream pathway of mTORC1 are 4E-binding proteins(4EBP) and phosphoprotein 70 S6Kinase(p70S6K). Some Chinese herbal extracts could improve cell proliferation via intervening the expressions of the key molecules in the upstream or downstream of PIK/Akt/mTOR signaling pathway in vivo. As for glomerular mesangial cells(MC) and podocyte, mTOR plays an important role in regulating glomerular inherent cells, including adjusting cell cycle, energy metabolism and matrix protein synthesis. Rapamycin, the inhibitor of mTOR, could suppress glomerular inherent cell hypertrophy, cell proliferation, glomerular basement membrane (GBM) thickening and mesangial matrix deposition in model rats with DN. Some Chinese herbal extracts could alleviate glomerular lesions by intervening mTOR signaling pathway activity in renal tissue of DN animal models or in renal inherent cells in vivo and in vitro.
Animals ; Diabetic Nephropathies ; drug therapy ; enzymology ; genetics ; pathology ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypertrophy ; drug therapy ; enzymology ; genetics ; pathology ; Kidney Glomerulus ; drug effects ; metabolism ; pathology ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; genetics ; metabolism

OBJECTIVETo evaluate clinical efficacy of suture anchors in treating acute injuries of medial collateral ligament (MCL) of knee at degree III.

METHODSTwenty-seven patients with degree III acute MCL injuries of knee were treated with suture anchors from January 2007 to June 2011. There were 15 males and 12 females, aged from 19 to 56 (averaged 32.6) years old. The time from injury to operation was 3 to 10 days, averaged 6 days. Symptoms and physical signs before and after treatment were observed, Lysholm scoring were used to evluated clinical efficacy.

RESULTSAll patients were followed up from 16 to 30 months with an average of 21.6 months. The stability of knee joints was good in all patients. Abduction stress test was negative when the knee joint was straightened at 0 degrees and flexed at 30 degrees. The average degree of flexed knee (67.00 +/- 5.80) degrees preoperatively was lower than that of postoperatively (136.50 +/- 6.30) degrees at 1 year. According to Lysholm scoring, preoperative scores ranged from 30 to 43 points, averaged 36.46 +/- 1.48; 1 year after operation ranged from 87 to 100 with an average of 91.50 +/- 3.80 and higher than postoperative. Twenty patients got an excellent results, 5 good and 2 fair.

CONCLUSIONSuture anchors in treating acute injuries of medial collateral ligament of knee at degree III has following advantages: small range of tissue dissection, easy to operate, reliable fixation and less complications.

Acute Disease ; Adult ; Collateral Ligaments ; injuries ; surgery ; Female ; Humans ; Knee Injuries ; surgery ; Male ; Middle Aged ; Suture Anchors

OBJECTIVETo establish a method for the simultaneous determination of 15 pesticides residues in Radix Paeoniae Alba by large volume injection-gas chromatography-mass spectrometry(LVI-GC-MS).

METHODSThe pesticides, including organochlorine pesticides, organophosphorus pesticides and pyrethroid insecticides, were analyzed by LVI-GC-MS using DB-5MS capillary column (30 m X 250 μm, 0.25 μm). The column temperature programming: initial temperature 40 degree for 1 min, with the increasing rate of 20 degree/min to 210 degree for 2 min, then with the increasing rate of 5 degree/min to 280 degree for 22 min. The flow of carrier gas was 1.0 ml/min with the injection volume of 15 μl.

RESULTSThe calibration curves of the pesticides were linear in the specified concentration ranges with correlation coefficients of 0.9937-0.9995. The average recoveries of the pesticides in Radix Paeoniae Alba spiked at two spiked levels ranged from 60.4 % to 106.8 % (for pendimethalin and 4, 4'-DDE those were 53.1 % and 45.2 %) with relative standard deviation(RSD) of 3.6 % to 18.6 % and the detection limits (S/N=3) were in the range of 0.16 μg/kg to 3.59 μg/kg.

CONCLUSIONThe established method for determination of multi-pesticide residue in Radix Paeoniae Alba is rapid, convenient and accurate with high sensitivity and low-cost.

Gas Chromatography-Mass Spectrometry ; methods ; Paeonia ; chemistry ; Pesticide Residues ; analysis

OBJECTIVETo investigate the safety and feasibility of dose dense chemotherapy in the postoperative adjuvant treatment for breast cancer.

METHODSThe data of 114 breast cancer patients treated with adjuvant dose dense chemotherapy was retrospectively analyzed.

RESULTS114 breast cancer patients received postoperative adjuvant dose dense chemotherapy from Jan. 2004 to March 2005. The patients were treated with different regimens according to their recurrence risk: 43 with paclitaxel + epirubicin (PE), 8 with taxotere + epirubicin (TE), 36 with cyclophosphamide + epirubicin + 5-Fu ( CEF) and 27 with epirubicin +5-Fu (EC). Totally, 111 patients except 3 completed the dose dense chemotherapy smoothly. Major toxicities severer than grade III included: neutropenia in 6.3% of the patients, nausea and vomit in 11.7%, peripheral neuropathy in 3.6%.

CONCLUSIONPostoperative adjuvant dose dense chemotherapy for breast cancer patient is feasible and safe, but the long-term outcome still needs further observation.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Breast Neoplasms ; drug therapy ; pathology ; surgery ; Chemotherapy, Adjuvant ; Cyclophosphamide ; administration & dosage ; Disease-Free Survival ; Dose-Response Relationship, Drug ; Epirubicin ; administration & dosage ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Humans ; Liver Neoplasms ; secondary ; Lymphatic Metastasis ; Mastectomy ; methods ; Middle Aged ; Nausea ; chemically induced ; Neutropenia ; chemically induced ; Paclitaxel ; administration & dosage ; Postoperative Period ; Retrospective Studies ; Taxoids ; administration & dosage

Public hospitals reform is a key roadblock for the ongoing health reform.By means of such experiments as Three openings and three mechanisms,Beijing is practicing a separation of hospital regulation and management and separation of clinic and pharmacy,while building the mechanism of financial subscription for pricing,that of medical insurance adjustment,and that of hospital corporate governance.These measures aim at building a new management structure,operation mechanism and medical service model focusing on quality of care,efficiency and satisfaction.Separation of clinic and pharmacy has lowered drug proportion,average outpatient expense and out of-pocket payment of patients,as well as producing higher patient satisfaction,quality of care and hospital income.Other benefits include better management efficiency indirectly caused by separation of clinic and pharmacy,higher acceptance of the corporate governance,and service model innovation to better serve the people.

Objective To compare the efficacy and safety of escitalo-pram combined with zolpidem and zolpidem alone on primary insomnia.Methods A total of 72 primary insomnia patients were recruited from outpatients in the Seventh Hospital of Hangzhou and were randomly divided into test group (n=36)and control group (n=36).Patients in the test group were treated with escitalopram 10-20 mg· d-1 combined zolpidem 5 -10 mg · d -1 for 8 weeks, while patients in control group were treated with zolpidem 5 -10 mg · d-1 only.The efficacy and ad-verse events were assessed with the pittsburgh sleep quality index (PSQI), patient health questionnaire(PHQ-9)and treatment emergent symptom scale ( TESS ) before and after 1, 4 and 8 weeks treatment.Results The score of PSQI and PHQ-9 in two groups were decreased significantly at week 1, 4, 8 after treatment(P<0.05).The scores of PSQI and PHQ-9 was lower in test group than that in control group at the end of 8 weeks after treatment( P<0.05).There were similar rates of adverse reactions between two groups ( P >0.05 ).Conclusion Escitalopram combined with zolpidem is significantly effective and safe for primary insomnia patients.

Objective To investigate the relationship between marital status and prevalence of peripheral arterial disease (PAD) among Beijing residents.Methods From May to August 2007,10 054 residents were interviewed to collect demographic and biochemistry information.Generalized linear mixed model was used to determine the relationship between marital status and prevalence of PAD in terms of OR and corresponding 95％ confidence intervals.Results After adjusting the traditional risk factors including age,area,lipid,serum glucose,blood pressure,central obesity,smoking,drinking and physical activities for PAD,relationship between marital status and prevalence of PAD was found both at the female and male groups of those with 45 years or older.Compared with married men,OR and 95％ confidence interval of PAD for unmarried men were 1.56 (0.39 - 6.35 ) and 1.61 (0.77 - 3.35 ) for those with 45 years younger and older respectively,while compared with married women,OR and 95％ confidence interval of PAD for unmarried women were 0.75 ( 0.22 - 2.57 ) and 1.78 ( 1.23 - 2.58 ) for those with 45 years younger and older respectively.Significant differences ( all P ＜ 0.01 ) were found between married and unmarried women of 45 years or older in terms of age,waist circumstances,systolic blood pressure,fasting glucose,total cholesterol,low-density lipoprotein and smoking.Conclusions Peripheral arterial disease was related to marital status for women of 45 years or older among Beijing residents.Estrogen level and the distribution of other PAD risk factors might contribute to the observed finding.