This article proposes the "Efficacy Theory" hypothesis of the traditional Chinese medicines (TCMs): TCMs take effects and weaken toxicities through the additive effects of numerous effective forms (including their constituents or/and metabolites) on a same target, the synergistic effects based on the overall action of the additive effects on individual targets and their toxicities scattering effects. A TCM may include approximately 1000 constituents and each constituent may produce about 100 metabolites in vivo after oral administration. Numerous effective forms of incalculable constituents and their metabolites could work like a "army group" together. When the quantity of a specific target molecule is larger than the pharmaceutical molecules, the molecules of different kinds of effective forms could combine with the target molecules successively, to exert the additive effects. When the target molecules are mostly occupied ("target most spaces occupied"), this TCM begins to work. The additive effects maybe exert not only in concentration but also in a time order way, which gives a sustained efficacy of TCM. The additive effects and the toxicities scattering effects are resulted from the same effective groups and not identical toxic groups among different effective form molecules. The "toxicities scattering effect" can be used to explain the non-toxic TCMs, but not fit for toxic TCMs. The efficacy theory showed that the variety of constituents and metabolites may participate in the process of pharmacodynamic actions, including the additive effects, synergy effects and toxicities scattering effects, which may be useful for explaining and developing the characteristic advantage of the TCMs. The questions we need to study or confirm are as follows: What are the TCMs' pharmacodynamic substance basis and mechanism made up of Why are toxicities of most TCMs' smaller How is the TCMs' "Efficacy Theory" which reflects characteristic advantage of TCMs applied in the research and development of new drugs.
Animals ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Medicine, Chinese Traditional ; Pharmaceutical Preparations ; chemistry

Acute Disease ; Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; Humans ; Infant ; Inpatients ; Insecticides ; poisoning ; Middle Aged ; Organophosphorus Compounds ; Pesticides ; poisoning ; Poisoning ; etiology ; mortality ; therapy ; Risk Factors ; Survival Rate

OBJECTIVETo study clinical value of shear wave elastography (SWE) in the evaluation of neck-shoulder myofascial pain syndrome.

METHODSFrom December 2013 to July 2014,30 patients diagnosed as neck-shoulder myofascial pain syndrome were in the treatment group,including 17 males and 13 females, with an average age of (44 ± 3) years old. Thirty healthy people were in the control group, including 22 males and 8 females, with a mean age of (37 ± 5) years old. The patients in the treatment group were treated with manipulation, once every other day, total 7 times. The SWE was used to detect tension part of trapezius muscle of patients in the treatment group before and after treatment, as well as to detect muscle belly at the descending part of trapezius muscle in the control group. The tissue elasticity and Yang's modulus value were recorded and compared.

RESULTSThe tissue elasticity chart of patients in the treatment group before treatment was mainly greenish blue with the score of 3.70 ± 1.53, and the Yang's modulus was (43.4 ± 15.6) kPa. The tissue elasticity figure after treatment was mainly blue with the score of 2.40 ± 0.87, and the Yang's modulus was (29.0 ± 5.9) kPa. Whereas in the control group, the tissue elasticity figure was mainly blue with the score of 1.60 ± 0.72, and the Yang's modulus was (24.0 ± 7.6) kPa. These were statistical differences between the two groups (P = 0.000).

CONCLUSIONSWE can be used as an evaluation method of manipulation treatment for neck-shoulder myofascial pain syndrome, which is an objective and sensitive detection method.

Adult ; Elasticity Imaging Techniques ; methods ; Female ; Humans ; Male ; Middle Aged ; Musculoskeletal Manipulations ; Myofascial Pain Syndromes ; diagnosis ; therapy ; Neck ; Shoulder

To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.
Amides ; chemistry ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Rhizome ; chemistry

This study is to develop an HPLC method for the simultaneous determination of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoisoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in Smilacis Glabrae Rhizoma. Samples of Smilacis Glabrae Rhizoma, Heterosmilacis Chinensis Rhizoma and Heterosmilacis Yunnanensis Rhizoma were separated on an Agilent Zorbax SB-C18 column with gradient elution of acetonitrile-0.05% phosphoric acid at a flow rate of 1.0 mL · min(-1). The detected wavelength was set at 230 nm and the column temperature was maintained at 35 °C. The contents of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in 84 Smilacis Glabrae Rhizoma samples were in the range of trace-1.381, trace-9.913, trace-3.673, 0.6706-27.08, trace-1.181, trace-4.833 and trace-2.754 mg · g(-1), respectively. The established method was used for analysis of common adulterants. The results demonstrated that the contents of (-)-epicatechin in Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma were 0.01163-0.06007 mg · g(-1) and 0.01583-0.08585 mg · g(-1), respectively, while the other six constituents were not detected. The method is simple and accurate, and can be used for the quality control of Smilacis Glabrae Rhizoma. The constituents of Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma are significantly different from Smilacis Glabrae Rhizoma, and they are not suitable for using as Smilacis Glabrae Rhizoma.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Liliaceae ; chemistry ; Rhizome ; chemistry

Objective To explore the effects of mesenchymal stem cells ( MSC ) treatment on platelet counts in mice with immune-mediated thrombocytopenia ( ITP) and the possible mechanism .Meth-ods ITP was induced by daily intraperitoneal injection of anti-platelet membrane CD 41 antibody (MWReg30) into BALB/c mice.The mice were then divided into experiment and control groups with 20 mice in each.Each mouse in experimental group was injected with 2×107 mesenchymal stem cells (MSC) through the tail vein .The numbers of blood platelets in mice from two groups were counted on days 5, 7 and 14 after MSC injection .Reverse transcriptase polymerase chain reaction ( RT-PCR) was performed to meas-ure T-bet and GATA-3 gene expression in peripheral blood mononuclear cells ( PBMCs ) at mRNA level on day 14.The levels of IFN-γ, IL-2, IL-4 and IL-10 in serum were detected by ELISA .Results The platelet counts in experimental group were significantly higher than those in control group on days 7 and 14 after MSC injection [(588.0±81.6)×109/L and (623.0±78.9) ×109/L vs.(317.0±90.1) ×109/L and (288.0± 87.8)×109/L ] (P<0.05).On day 14 after MSC injection, the T-bet expression at mRNA level in PBMCs from mice in experimental group was significantly lower than that in control group [(0.04±0.03) vs.(0.27 ±0.05)] (P<0.05), while the GATA-3 expression at mRNA level was higher than those in control group [ (0.14±0.04) vs.(0.07±0.05)] (P<0.05).Compared with control group, the concentrations of Th1 type cytokines such as IFN-γand IL-2 were remarkably down-regulated in experimental group [(3.1±1.7) pg/ml and (3.2±2.1) pg/ml vs.(10.3±4.8) pg/ml and (16.3±5.7) pg/ml](P<0.05), while the con-centrations of Th2 type cytokines such as IL-4 and IL-10 were up-regulated in experimental group [(88.6± 15.2) pg/ml and (38.3±11.8) pg/ml vs.(32.7±5.7) pg/ml and (22.1±3.4) pg/ml ] (P<0.05). Conclusion MSC treatment can effectively increase platelet counts in mice with immune-mediated thrombo-cytopenia, which may be associated with the suppression of Th 1-dominant response mediated by abnormal ex-pression of T-bet and GATA-3.

0.05 for all).Conclusions The allelic frequencies of LPL gene at Pvu Ⅱ locus in Hei Yi Zhuang were different from those in Han,but the genotypie frequencies in Hei Yi Zhuang were not different from those in Han.There was no significant correlation between the polymorphism of LPL gene at Pvu Ⅱ site and the serum lipid levels in two ethnic groups.

Streptomyces hygroscopicus 17997 produces the antiviral and antitumor ansamycin antibiotic, geldanamycin. Studies on geldanamycin biosynthetic pathway will provide good tools for genetic manipulation of the antibiotic-producing strain to improve the productivity or to facilitate making novel geldanamycin analogs. The structural similarities between geldanamycin and ansamycins such as rifamycin or ansatrienin suggest that both geldanamycin and ansamycins has a closely related pathways of biosynthesis and that biosynthetic system for geldanamycin is similar to the one of type I polyketide synthase (PKS) enzyme system. To explore the possible PKS genes involved in geldanamycin biosynthesis, the degenerate primers were designed according to the conserved sequence of KS-AT region from erythromycin and oleandomycin type I PKS genes. Cosmids containing multiple PKS genes (pCGBK2,4,6,10,11,18) were obtained by hybridization with the PCR products, which were amplified from S. hygroscopisus 17997 genomic DNA. The designed primers above were used for PCR. Development of a Streptomyces temperate phage phiC31-derivative KC515( tsrR) transduction system was carried out for identification of cosmids containing the PKS gene related to biosynthesis of geldanamycin. Several factors, mainly the Ca2+ and Mg + concentrations in different culture media affecting the frequency of gene transfection, were optimized .Transfection efficiency could reach up to 10(3) /microg DNA on YMG medium supplemented with 10mmol/L MgSO4. Reversely, the transfection efficiency decreased when YMG medium was supplemented with 30mmol/L MgSO4. Gene transfection system based on the integration-defective phage KC515 had been established for S. hygroscopicus17997. Recombinant phages (ph111, 258, 287, 116, 105) were constructed by insertion of the homologous to PKS gene fragments into the KC515 phage vector. Gene disruption experiments were performed by transduction of recombinant phages into S. hygroscopicus 17997 genome, and disruption of geldanamycin production was observed as a result of homologous recombination between the cloned insert in recombinant phage and the S. hygroscopicus 17997 genome by integration. Thiostrepton resistant transductants were selected and integration event was analyzed by Southern hybridization. The fermentation broth extracts from five resistant transductants were analyzed by TLC and HPLC. The results showed that only G16 mutant failed to produce geldanamycin. This result showed that the integration of the insert DNA fragment in recombinant phage phl6 into the chromosome of S. hygroscopicus disrupted the expression of the geldanamycin biosynthetic genes. The original cosmid pCGBK10 containing this cloned insert was predicted to encode PKS genes in the geldanamycin biosynthesis. This study laid the foundation for cloning the PKS genes involved in geldanamycin biosynthetic gene cluster from S. hygroscopicus 17997.
Alcohol Oxidoreductases ; genetics ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Bacteriophages ; genetics ; Benzoquinones ; metabolism ; Blotting, Southern ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Genetic Vectors ; genetics ; Lactams, Macrocyclic ; metabolism ; Multigene Family ; genetics ; physiology ; Polymerase Chain Reaction ; Streptomyces ; genetics ; metabolism

Objective:To explore the influencing factors for postoperative delirium in General Ward of Neurosurgery and evaluate the influence of serum acetylcholinesterase level in it.Methods:A retrospective study was performed. Two hundred and ninety-eight patients accepted surgery and diverted into General Ward of Neurosurgery in our hospital from January 2021 to July 2021 were chosen in our study. The 4AT delirium scoring tool was used to evaluate whether the patients had delirium, and these patients were, then, divided into non-delirium group and delirium group. The preoperative general data, history of deseases and laboratory results (serum acetylcholinesterase level) were collected. Univariate analysis and multivariate Logistic regression analysis were used to determine the independent factors affecting the occurrence of postoperative delirium, especially the relation between preoperative serum acetylcholinesterase level and postoperative delirium. Receiver operating characteristics (ROC) curve was drawn to evaluate the predictive value of serum acetylcholinesterase in postoperative delirium.Results:The incidence of postoperative delirium in 298 patients in General Ward of Neurosurgery was 24%, including 225 patients into the non-delirium group and 73 patients into the delirium group. There were significant differences between the two groups in the proportions of patients having resuscitation in anesthesia ICU, using postoperative analgesic pump and having alcoholism history, surgical duration, intraoperative bleeding, proportion of patients accepting skull base surgery, proportion of patients remaining awake 2 h after surgery, and incidence of bilateral frontal lobe pneumatosis after surgery ( P<0.05). Preoperative serum acetylcholinesterase level in delirium group ([2.35±0.49] U/mL) was significantly lower than that in non-delirium group ([2.78±0.48] U/mL, P<0.05). Preoperative serum acetylcholinesterase level ( OR=0.116, 95%CI: 0.034-0.394, P=0.001), postoperative resuscitation in anesthesia ICU ( OR=0.043, 95%CI: 0.002-0.878, P=0.041), keeping awake 2 h after surgery ( OR=7.641, 95%CI: 1.675-34.858, P=0.009), surgical duration ( OR=1.887, 95%CI: 1.192-2.987, P=0.007), intraoperative bleeding ( OR=1.010, 95%CI: 1.006-1.014, P<0.001), and skull base surgery ( OR=6.700, 95%CI: 1.907-23.547, P=0.003) were all independent influencing factors for postoperative delirium in patients in General Ward of Neurosurgery. The area under ROC curve for serum AchE level to predict the occurrence of postoperative delirium was 0.735(95%CI: 0.679-0.800, P<0.001); when the cut-off value was 2.67 U/mL, the sensitivity and specificity were 64% and 75%. Conclusions:Skull base surgery, keeping awake 2 h after surgery, long surgical duration and large amount of intraoperative bleeding can promote the occurrence of postoperative delirium; admission to anesthesia ICU after surgery can reduce the occurrence of delirium. When the preoperative serum AchE level is less than 2.67 U/mL, the possibility of postoperative delirium should be warned.

BACKGROUNDTreponema pallidum (T. pallidum) subsp. pallidum is the causative agent of syphilis. Analysis of recombinant antigens of T. pallidum led to the identification of potential candidate antigens for vaccine development and syphilis serodiagnosis. Tp0965 was predicted to be a membrane fusion protein and was found to be reactive with infected human sera in previous studies, but the results were controversial. In this research, the antigenicity and immunoreactivity of recombinant protein Tp0965 were assessed.

METHODST. pallidum subsp. pallidum (Nichols strain) was propagated and isolated and the genomic DNA was extracted. The Tp0965 gene was amplified by polymerase chain reaction (PCR). Then the recombinant protein Tp0965 was expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid (Ni-NTA) purification system. The reactivities of protein Tp0965 were examined by immunoblot analysis and indirect enzyme-linked immunosorbent assay. The antisera against protein Tp0965 were obtained by immune rabbits and the immunogenicity of antisera were detected by indirect enzyme-linked immunosorbent assay.

RESULTSRecombinant protein Tp0965 was expressed successfully in vitro. Immunoblot assay showed that the recombinant protein Tp0965 could be recognized by human syphilitic sera of all stages. Indirect enzyme-linked immunosorbent assay showed there were only 4 of 74 human syphilitic sera that failed to show reactivity to recombinant antigen Tp0965, and lack of reactivity of Tp0965 to all 28 uninfected sera. A low titer of antiserum against Tp0965 in immune rabbits could be detected after the third time of immunization.

CONCLUSIONSThe recombinant antigen Tp0965 shows excellent sensitivity for the reactivity with sera from syphilitic individuals at all stages. The results also demonstrate a potential application for the serodiagnosis of syphilis.

Animals ; Antigens, Bacterial ; genetics ; immunology ; Bacterial Proteins ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Membrane Proteins ; genetics ; immunology ; Polymerase Chain Reaction ; Rabbits ; Syphilis ; immunology ; microbiology ; Treponema pallidum ; immunology ; metabolism