This paper briefly analyzed and discussed the current status and major scientific challenges of traditional Chinese medicine (TCM) formulaology research. To promote formulaology research, a new strategy and corresponding technology, network formulaology, were proposed to reveal the complex interaction between functional chemome and biological responses network. The research framework and directions of network formulaology were also summarized and prospected.
Chemistry, Pharmaceutical ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; Internet ; Medicine, Chinese Traditional ; standards

Objective To prepare recombinant human prothrombin-2 expressed in Pichia pastoris, and assay the enzymatic and clotting activities of prothrombin-2 activated by prothrombin activator ecarin.Methods Human prothrombin-2 gene and Echis carinatus ecarin gene were synthesized separately on the basis of the cDNA sequences published in GenBank.The gene of prothrombin-2 was cloned into the expression vector pPICZαA.The expression vector pPICZαA/prothrombin-2 was transformed into glycoengineered P.pastoris, and then prothrombin-2 engineered P.pastoris was screened.The expression products were induced by methanol, purified by two-step chromatography and identified by diges-tion by PNGase F and analysis of pepetide fingerprint.The ecarin gene was cloned into the expression vector pcDNA3.1. The expression vector pcDNA3.1/Ecarin was transformed into HEK 293T cells and the culture supernatant of HEK 293T/Ecarin was collected.The reaction product of HEK 293T/Ecarin cell culture supernatant and purified prothrombin-2 was analyzed by S-2238,which was the chromogenic substrate for thrombin.Fibrinogen was used to measure blood clotting time. Results The purified protein of P.pastoris expressed prothrombin-2 culture supernatant was 37 ×103 .The relative molecular mass(Mr) of the purified protein was reduced to 35 ×103, which was consistent with the theoretical Mr of prothrombin-2 molecular weight.The purified protein was proved to be prothrombin-2 by peptide fingerprint identification. The purified prothrombin-2 processed by HEK 293T/Ecarin culture supernatant could hydrolyze S-2238 to produce yellow pNA, and D405 of pNA increased with the volume of the processed prothrombin-2 that could promote the plasma coagulation.The blood clotting time was close to that of the thrombin kit.Conclusion Prothrombin-2 is prepared by P.pastoris and activated toα-thrombin by ecarin.This technique may replace the method of extraction of prothrombin from plasma and can be used for the treatment of war wounds or for future clinical research.

Objective Alstr(o)m syndrome (AS) is a rare,autosomal recessive inherited disease characterized by various clinical manifestations.The aim of this study was to review the clinical characteristics and laboratory findings of AS.Methods Two cases of AS was reported.Combined with the clinical data of 7 cases of AS which had been reported in China,the clinical characteristics and laboratory findings of AS were reviewed.Results Visual disorder( median onset age:6.0 years ) and dysaudia( median onset age:10.3 years ) were found in 9 patients,short stature and obesity in 8 patients,acanthosis nigricans in 7 patients,diabetes mellitus( median onset age:14.5 years) in 6 patients,and heart disease in 4 patients; hyperuricemia was detected in 6 patients,hepatic dysfunction and hypertriglyceridemia in 5 patients.Conclusions Visual disorder was the first presentation in patients with AS.Deafness,obesity,diabetes,and short stature were common.These findings were helpful in making an early and accurate diagnosis and appropriate treatment.

Objective To observe the improvement of postoperative pulmonary function and oxygen partial pressure during general anesthesia for open abdominal surgery with lung protective ventilation strategies and alveolar recruitment maneuvers. Methods Seventy patients who underwent selective open abdominal surgery were selected, and they were divided into standard ventilation group (tidal volume 8 ml/kg) and protective ventilation group (tidal volume 6 ml/kg, 5 cmH2O positive end-expiratory pressure, and alveolar recruitment maneuvers, 1 cmH2O=0.098 kPa) according to the random digits table method with 35 cases each. The airway pressure, blood pressure, pulse oxygen saturation (SpO2), end-tidal partial pressure of carbon dioxide (PETCO2) and adverse reactions were observed. The SpO2, partial pressure of O2 (PaO2) and pulmonary function before surgery and 1, 3, 5 d after surgery were measured. Results The respiratory rate, airway pressure and PETCO2 levels in protective ventilation group were significantly higher than those in standard ventilation group: (12.3 ± 2.1) times/min vs. (10.2 ± 1.0) times/min, (15.1 ± 2.8) cmH2O vs. (13.5 ± 2.3) cmH2O, (34.6 ± 2.1) mmHg (1 mmHg=0.133 kPa) vs. (32.1 ± 1.4) mmHg, and there were statistical differences (P<0.05). The SpO2 in 2 groups was maintained at 0.99. There was no statistical difference in the incidence of postoperative complications between 2 groups (P>0.05). The SpO2 and PaO2 levels at 1, 3 d after surgery in protective ventilation group were significantly higher than those in standard ventilation group:0.951 ± 0.018 vs. 0.936 ± 0.016 and 0.964 ± 0.018 vs. 0.949 ± 0.018, (74.8 ± 6.8) mmHg vs. (65.0 ± 6.2) mmHg and (79.6 ± 6.0) mmHg vs. (70.6 ± 5.3) mmHg, and there were statistical differences (P<0.05). The forced expiratory volume in 1 s (FEV1), percentage of the estimated value of FEV1, forced vital capacity (FVC) and percentage of the estimated value of FVC at 1, 3 and 5 d after surgery in protective ventilation group were significantly higher than those in standard ventilation group, the FEV1/FVC at 1 d after surgery was significantly higher than that in standard ventilation group, and there were statistical differences (P<0.05). Conclusions The lung protective ventilation strategy and alveolar recruitment maneuvers can improve the postoperative pulmonary function and oxygen partial pressure during general anesthesia for abdominal surgery. Low vital volume, appropriate positive end-expiratory pressure and recruitment maneuvers can protect the lung in general anesthesia patients.

Objective To investigate the role of receptor for advanced glycation end products (RAGE)/NF-κB signaling pathway in mediating lysophosphatidylcholine (LPC)-induced TGF-β1 expression in human retinal endothelial progenitor cells (HEPCs).Methods Human retinal endothelial cells (HERCs)were transfected with siRNA for RAGE siRNA or added NF-κB in-hibitor pyrrolidine dithiocarbamate (PDTC)in the presence or absence of LPC,the expressions of TGF-β1 and RAGE genes were analyzed by qPCR and Western blot.Results LPC could increase the expression of RAGE and TGF-β1 gene in HERCs.The RAGE gene after silence could significantly decrease the expression of LPC-induced RAGE and TGF-β1 .Adding NF-κB inhibitor PDTC significantly reduced LPC-induced RAGE and TGF-β1 expression in HERCs.Conclusion RAGE/NF-κB signaling pathway plays an important role in mediating LPC induced TGF-β1 gene expression in HERCs.

Objective To investigate the association of vitamin D receptor gene (Apa1,Bsm1,Taq1) polymorphisms with autoimmune liver diseases risk.Metiods Case control test documents were retrieved through Pubmed,Ovid,Medline,and Web of science databases,according to the inclusion and exclusion criteria included in this study.The design of experiments,the characteristics of the object of study,research results was excerpted,STATA version 12.0 software were used.The correlation intensity was demonstrated with odds ratio (OR) and 95％ confidence interval (CI).Results A total of 6 publications containing 9 studies (7 studies about primary biliary cirrhosis,2 studies about autoimmune hepatitis) published from January 2000 to February 2012 were identified and 844 cases and 1 522 controls were included.The combined results based on all studies showed that there was a statistically significant link between Apa 1 and autoimmune liver diseases (OR =0.85,95％ CI 0.74-0.96,P =0.058,for a vs.A; OR =0.75,95％ CI 0.58-0.97,P =0.212,for aa vs.AA;OR =0.78,95％ CI 0.63-0.98,P =0.235,for Aa vs.AA; OR =0.77,95％ CI 0.63-0.94,P =0.231,for Aa/aa vs.AA),while the Bsm 1 and Taq 1 didn' t show the association with autoimmune liver diseases.Conclusion The current meta-analysis shows that Apal may be a low-penetrant risk factor for autoimmune liver diseases.Bsm1 and Taq1 don't show the association with autoimmune liver diseases.

OBJECTIVE:To establish a method for simultaneous determination of spironolactone and erythromycin in Com-pound spironolactone gel. METHODS:HPLC was performed on the column of Thermo-Hypersil ODS2-C18 with mobile phase of 0.1 mol/L ammonium dihydrogen phosphate solution (pH was adjusted to 7.0 by triethylamine)-acetonitrile (60∶40,V/V) at flow rate of 1.0 ml/min,detection wavelength was 215 nm and 238 nm,column temperature was 30℃,and injection volume was 5 μl. RESULTS:The linear range was 0.251 6-5.032 μg/ml for spironolactone and 0.577 2-11.544 μg/ml(r=0.999 9) for erythromycin (r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 0.83%;average recoveries were 97.8%(RSD=0.74%,n=9)and 96.7%(RSD=2.60%,n=9),respectively. CONCLUSIONS:The method is simple,reproducible,ac-curate and reliable,and can be used for the quality control of Compound spironolactone gel.

Objective To explore the mechanism of LMWH therapy for SAP.Methods 48 wistar rats were random divided into 3 groups,sham group(S group),severe acute pancreatitis group(SAP group)and LMWH therapy group(H group).Serum amylase,IL-6,acinar cell apoptosis and the activity of NF-κB were detected and compared.Results The expression of amylase and IL-6 in SAP group was significantly higher than that in H group(P＜0.01).The apoptosis index of acinar cell in SAP group wag significantly lower than that in H group(P＜0.01),while the activity of NF-κB in SAP groupwas stronger than that in H group.Conclusions LMWH therapy may ameliorate SAP by inducing acinar cell apoptosis through suppressing the activity of NF-κB.

Objective To investigate the mechanism of preventing islet β-cell apoptosis in NOD mice with pioglitazone.Methods Female NOD mice at 4 weeks of age were divided into pioglitazone group ( n =21,0.02％pioglitazone was added into the feed ) and control group ( n =21,fed with regular diet).The accumulative incidence of diabetes was followed-up to 52 weeks of age in each group of NOD mice.Pancreas was removed from NOD mice at 12 weeks of age in each group ( n =15 ) to score severity of insulitis by routine H-E staining.The apoptotic β-cells in islets were observed with double-labeling technique of TUNEL in situ combined with standard sensitive avidin-biotin complex (sABC) immunohistochemical method.The spleens were taken for cell culture; IL-4 and IFN-γ levels in sera and supernatants of cultured splenocyte,the activity of PPARγ and NF-κB nuclear proteins in cultured splenocyte were measured by ELISA.Results (1)At 30 and 52 weeks of age,the respective incidences of diabetes were 57.1％ and 76.2％ in pioglitazone group,and 76.2％ and 90.5％ in control group ( all P＞0.05 ).At 15 weeks of age,the incidence became 4.8％ in pioglitazone group,and 33.3 ％ in control group ( P =0.045 ).( 2 ) At 12 weeks of age,the percentages of non infiltrated islet and peri-insulitis islet in pioglitazone group were higher than those in control group ( 14.73％ vs 5.69％,P＜0.01 ; and 26.02％ vs 15.72％,P＜0.01 ),and that of intraislet insulitis was lower than that in control group ( 59.25％ vs 78.59％,P＜0.01 ).The percentage of apoptotic β-cell in pioglitazone group was lower than that in control group( 6.17％ ±3.62％ vs 10.62％ ±4.43％,P=0.008 ).(3) In sera,IFN-γ level in pioglitazone group was lower than that in control group [( 561.05±78.61 ) vs ( 666.43 ± 28.42 ) pg/ml,P =0.045].In cultured splenocyte supernatant,the level of IFN-γ in pioglitazone group was lower than that in control group[(605.84+65.60) vs (692.20+44.98) pg/ml,P=0.041].(4) In cultured splenocyte,PPARγ nuclear protein activity in pioglitazone group was higher than that in control group ( 0.06 ± 0.01 vs 0.03 ± 0.01,P =0.013 ),and NF-κB nuclear protein activity was lower than that in control group ( 0.03 ± 0.01 vs 0.08± 0.01,P =0.001 ).Conclusions Pioglitazone activates PPARγ nuclear protein,inhibits activity of NF-κB nuclear protein,downregulates IFN-γ,diminishes differeutiation of Th cells to Th1,and subsequently prevents insulitis and β-cell apoptosis in NOD mice.

This study aims to provide an effective theory-based intervention model on health education for osteoporosis in accordance with the International Classification of Functioning, Disability and Health (ICF) theory of functioning and disability. The framework and approach,significance and contents had been discussed