This article introduced professor WANG Shu-chen's experience in picture treating bronchiolitis obliterans syndrome (BOS) after hematopoietic stem cell transplantation with tougue picture as the key link. Patients with BOS showed deficiency of essence of tongue crack, which should be treated by warming kidney yang, replenishing essence and marrow; fester tongue for qi disorder, inflammation caused by fire, which should be treated by regulating qi, and clearing heat; exfoliative fur accumulation, which should be treated with blood stasis and toxin, removing blood stasis and toxin; thick and greasy fur, which should be treated by warming spleen and activating spleen.

Objective:To systematically evaluate the therapeutic efficacy of tuina therapy for primary insomnia.Methods:Nine Chinese and English databases were searched from the inception to May 2017 to identify randomized controlled trials (RCTs) studying tuina therapy for insomnia.The enrolled articles were all RCTs with tuina as the monotherapy or major therapy in the experiment group,with clear diagnostic criteria for primary insomnia well recognized worldwide or in China,and Pittsburgh sleep quality index (PSQ I) as one of the outcome measures.Two researchers evaluated the risk of bias and quality of the enrolled studies by following Cochrane Handbook version 5.1.0.The meta-analysis was performed by RevMan version 5.3.Results:Eleven studies were included with a total of 1 076 participants.The Western medication adopted in the control groups were benzodiazepine receptor agonists.The studies were all assessed as high risk of bias for blinding since blinding method was unable to be performed due to the specificity of tuina therapy;no study reported the support of fund or potential interest conflict,so they were all rated unclear for selective reporting.The meta-analysis showed that compared with other traditional Chinese medicine therapies,tuina worked more effectively in reducing the PSQI score (MD=-4.11＜0,95％ confidence interval (CI)-6.01 to-2.22,P＜0.0001);compared with oral administration of Western medication,tuina showed more significant efficacy in reducing the PSQI score (MD=-3.42＜0,95％CI-5.19 to-1.66,P＜0.0001).Subgroup analysis showed that head tuina alone showed no significant difference compared with oral administration of Western medication regarding the change of PSQI score (MD=-4.19＜0,95％CI-8.87 to 0.50,P＞0.05);a combination of head and back tuina could more effectively reduce the PSQI score compared with oral administration of Western medication (MD=-2.08＜0,95％CI-3.09 to-1.06,P＜0.0001).Conclusion:Tuina can produce more significant efficacy in treating primary insomnia compared with other traditional Chinese medicine therapies and oral administration of Western medication,especially the combination of head and back tuina.

The use of monoclonal antibodies (mAbs) for cancer therapy has achieved considerable success in recent years. Approximate 17 monoclonal antibodies have been approved as cancer therapeutics since 1997. Antibody-drug conjugates (ADC) are powerful new treatment options for cancer, and naked antibodies have recently achieved remarkable success. The safety and effectiveness of therapeutic mAbs in oncology vary depending on the nature of the target antigen and the mechanisms of tumor cell killing. This review provides a summary of the current state of antibody-based cancer therapy, including the mechanisms of tumor cell killing by antibodies, tumor antigens as antibody targets, clinical effectiveness of antibodies in cancer patients and nanoparticles-based ADCs.
Antibodies, Monoclonal ; immunology ; therapeutic use ; Antigens, Neoplasm ; immunology ; Antineoplastic Agents ; therapeutic use ; Humans ; Immunoconjugates ; therapeutic use ; Nanoparticles ; Neoplasms ; immunology ; therapy

OBJECTIVEN-acetyltransferase 2 (NAT2) and cytochrome P450 2EI (CYP2E1) play a crucial role in the drug metabolic process. The aim of this study was to understand the genotype and phenotype polymorphisms of NAT2 and CYP2E1 in the Han Chinese pediatric population in order to provide a theoretical basis for individualized drug treatment.

METHODSA total of 341 (211 males and 130 females) randomly sampled Han Chinese children, aged from 2 months to 14 years, were enrolled in this study. Genotyping was carried out by PCR method, and metabolic phenotypes were identified.

RESULTSIn this study population, wild genotype was found as a major genotype in seven SNPs of NAT2, rs1801279, rs1041983, rs1801280, rs1799929, rs1799930, rs1208 and rs1799931. The frequency of NAT2 fast metabolism was highest (61.3%), followed by middle to slow metabolism (34.1%). Wild genotype also predominated in the four SNPs of CYP2E1 (rs2031920, rs3813867, rs6413432 and rs72559720) named as CYP2E1*5, *6 and *2, with a frequency of 61.3%, 60.1% and 99.4% respectively. As the relationship between CYP2E1 genotype and phenotype was unknown, phenotyping of CYP2E1 was not done.

CONCLUSIONSThe important SNPs of NAT2 and CYP2E1 are predominantly wild genotype in the Han Chinese pediatric population. Fast metabolic phenotype predominates in important SNPs of NAT2.

Adolescent ; Arylamine N-Acetyltransferase ; genetics ; Child ; Child, Preschool ; China ; ethnology ; Cytochrome P-450 CYP2E1 ; genetics ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Male ; Phenotype ; Polymorphism, Single Nucleotide

The paper is aimed to study the dynamic accumulation regulation of curcumin (Cur), demethoxycurcumin (DMC) and bisdemethoxyeurcumin (BDMC) in three strains of Curcuma longa, and provide scientific references for formalized cultivation, timely harvesting, quality control and breeding cultivation of C. longa. The accumulation regulation of the three curcumin derivatives was basically the same in rhizome of three strains. The relative contents decreased along with plant development growing, while the accumulation per hectare increased with plant development growing. The accumulation of curcuminoids per hectare could be taken as the assessment standard for the best harvest time of C. longa. A3 was the best strain in terms of Cur and BDMC content.
Curcuma ; chemistry ; growth & development ; metabolism ; Curcumin ; analogs & derivatives ; analysis ; metabolism ; Quality Control ; Rhizome ; chemistry ; growth & development ; metabolism

Recent studies have shown that epidermal growth factor receptor (EGFR) is an important target for cancer therapy. The present study prepared single chain Fv (scFv) directed against EGFR. Balb/c mice were immunized by human carcinoma A431 cells, and total RNA of the splenic cells was extracted. VH and VL gene fragments were amplified by RT-PCR and further joined into scFv gene with a linker, then scFv gene fragments were ligated into the phagemid vector pCANTAB 5E. The phagemid containing scFv were transformed into electro-competent E. coli TG1 cells. The recombinant phage antibody library was constructed through rescuing the transformed cells with help phage M13K07. The specified recombinant phages were enriched through 5 rounds of affinity panning and the anti-EGFR phage scFv clones were screened and identified with ELISA. A total of 48 clones from the library were selected randomly and 45 clones were identified positive. After infecting E. coli HB2151 cells with one positive clone, soluble recombinant antibodies about 27 kD were produced and located in the periplasm and the supernatant. The result of sequencing showed that the scFv gene was 768 bp, which encoded 256 amino acid residues. VH and VL including 3 CDRs and 4 FRs, respectively, were all homologous to mouse Ig. The soluble scFv showed the specific binding activity to purified EGFR and EGFR located in carcinoma cell membrane. The successful preparation of anti-EGFR scFv will provide an EGFR targeted molecule for the development of antibody-based drugs and biological therapy of cancer.
Animals ; Antibodies, Monoclonal ; Cell Line, Tumor ; Humans ; Immunoglobulin Light Chains ; genetics ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Peptide Library ; Receptor, Epidermal Growth Factor ; genetics ; immunology ; Single-Chain Antibodies ; genetics ; immunology

AIMTo analyze the chemical components in Danggui (the roots of Angelica sinensis (Oliv.) Diel).

METHODSHPLC-MS/MS was used to identify the main components in Danggui. Furthermore, the MS fragmentation regularity of the phthalides was proposed. The mobile phase of HPLC consisted of 0.5% acetic acid in water and 0.5% acetic acid in acetonitrile, analytical column was Hypersil ODS2 (250 mm x 4.6 mm, 5 microm), flow rate 1.0 mL x min(-1), injected volume 2 microL. The ionization source was ESI in positive ion mode.

RESULTSFerulic acid, nine known phthalides and one unknown phthalide derivative were tentatively identified in chromatograms based on their MS data and the comparison of their UV spectra with those published in the literatures.

CONCLUSIONThe structural information of phthalides was obtained via HPLC-MS/MS, which provides an accurate and fast method to identify the phthalides and provides more scientific information for quality control of Danggui.

4-Butyrolactone ; analogs & derivatives ; analysis ; chemistry ; Angelica sinensis ; chemistry ; Benzofurans ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; chemistry ; Molecular Structure ; Phthalic Anhydrides ; analysis ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Spectrometry, Mass, Electrospray Ionization ; methods

Carboxypeptidase B is a metalloenzyme, which is widely used for commercial and research purposes. Commercially available CPB purified from porcine or bovine pancreas is very expensive, and is not totally free from other proteases. In order to express the rat proCPB in Pichia pastoris, total RNA extracted from SD rat pancreas cells was reversely transcripted to synthesize cDNA, and the proCPB ORF was synthesized by PCR. After digestion with Xho I and EcoR I , the fragment was inserted into pPIC9, and the recombinant plasmid was named as pPIC9-proCPB. By digestion with Sac I , the lined pPIC9-proCPB was transformed into Pichia pastoris strains GS115 with PEG1000 and integrated into their genomes. In the inducement of methanol, recombinant proCPB was successfully expressed in Pichia pastoris, and could be secreted into the supernatant in the culture. After optimizing the fermentation conditions, a higher production could be obtained when GS115-proCPB was induced in BMGY (pH6.0) at 28CC, with addition of 0.5% casein. The yield of recombinant protein reached 500mg/L, achieving over 94% of total protein in the culture supernatant. The purity of recombinant CPB can reach 96% after two step phenyl sepharose F F purification, and 38% of total protein can obtained after optimizing the pufication method. Comparing to the specific activity 180u/mg of CPB purchased from Sigma, the specific activity of recombinant CPB is 110u/mg. Mass spectrometry analyses showed the mass of the recombinant CPB was 35.1 kD, which is very close to the theory value 35.2 kD. Amino acid sequencing of N-terminal of recombinant CPB further indicated proCPB was expressed successfully and modificated correctly after translation.
Animals ; Carboxypeptidase B ; genetics ; isolation & purification ; metabolism ; Catalysis ; Cattle ; Electrophoresis, Polyacrylamide Gel ; Gene Expression Regulation, Enzymologic ; Kinetics ; Mass Spectrometry ; Molecular Weight ; Pichia ; genetics ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; chemistry ; isolation & purification ; metabolism ; Sequence Analysis, Protein ; Substrate Specificity ; Swine

Objective To construct RNAi combinant adenoviral expressive vectors specific to p65 subunit of NF-?B and to observe their gene silencing effect on p65 subunit.Methods Three pairs of complementary. single-strand DNA oligos targeting three various sites of p65 mRNA were designed and synthesized.Annealling was used to generate double-strand oligos(ds-oligos),and then the ds-oligos were cloned into pENTR~TM/u6 to generate the entry clone named pENTR.Recombination reaction in vitro with the pENTR and pAd/BLOCK-iT~TM- DEST was used to creat the adenovirus plasmid which contains the RNAi cassette.Then,the adenovirus plasmids digested with PacI were transfected into HEK293A cells to product adenovirus,and latter infected the HEK293A cells to amplify the adenoviral stock.Plaque forming assay was used to titer the adenoviral stock.The p65 gene silencing effect induced by the RNAi adenovirus was detected by Western blot and immunocytochemistry assay in ECV304 cells.Results The RNAi adenovirus specific to p65 subunit of NF-?B were produced with titer of 3.0 x 10~9pfu/ml to 2.5?10~10pfu/ml.The expression of p65 protein in ECV304 cells could be down-regulated efficiently by the RNAi adenovirus 48-72 h after infection,which would last for more than 6 days after infection.Conclusion RNAi adenovirus is an important tool inhibiting the expression of target gene efficiently.

China implemented the public hospital reform in 2012.This study utilized bootstrapping data envelopment analysis (DEA) to evaluate the technical efficiency (TE) and productivity of county public hospitals in Eastern,Central,and Western China after the 2012 public hospital reform.Data from 127 county public hospitals (39,45,and 43 in Eastern,Central,and Western China,respectively) were collected during 2012-2015.Changes of TE and productivity over time were estimated by bootstrapping DEA and bootstrapping Malmquist.The disparities in TE and productivity among public hospitals in the three regions of China were compared by Kruskal-Wallis H test and Mann-Whitney U test.The average bias-corrected TE values for the four-year period were 0.6442,0.5785,0.6099,and 0.6094 in Eastern,Central,and Western China,and the entire country respectively,with average non-technical efficiency,low pure technical efficiency (PTE),and high scale efficiency found.Productivity increased by 8.12％,0.25％,12.11％,and 11.58％ in China and its three regions during 2012-2015,and such increase in productivity resulted from progressive technological changes by 16.42％,6.32％,21.08％,and 21.42％,respectively.The TE and PTE of the county hospitals significantly differed among the three regions of China.Eastern and Western China showed significantly higher TE and PTE than Central China.More than 60％ of county public hospitals in China and its three areas operated at decreasing return scales.There was a considerable space for TE improvement in county hospitals in China and its three regions.During 2012-2015,the hospitals experienced progressive productivity;however,the PTE changed adversely.Moreover,Central China continuously achieved a significantly lower efficiency score than Eastern and Westem China.Decision makers and administrators in China should identify the causes of the observed inefficiencies and take appropriate measures to increase the efficiency of county public hospitals in the three areas of China,especially in Central China.