Advances of high-throughput screening system in reengineering of biological entities / 生物工程学报

Enzymes and cell factories are the core of industrial biotechnology. They play important roles in various fields such as medicine, chemical industry, food, agriculture, and energy. Usually, natural enzymes and cells need to be engineered to improve the catalytic efficiency, stability and enantioselectivity. Directed evolution makes it possible to rapidly improve the properties of enzymes and cell factories. Sensitive and reliable high-throughput screening approaches are the key for successful and efficient engineering of enzymes and cell factories. In this review, we first summarize the advantages and disadvantages of different screening methods and signal generation strategies as well as their application scope; we then describe the latest advances of ultra-high throughput screening technology applied in the directed evolution of enzymes and cell factories in the past three years. On this basis, we discuss the limiting factors that need to be further improved for high-throughput screening systems and forecast the future development trends of high-throughput screening methods, hoping that researchers in various fields including biotechnology and instrument development can cooperate closely to enhance the reliability and applicability of the high-throughput screening techniques.

An empirical study on the evaluation system of clinical nurse training in hospitals of traditional Chinese and Western medicine based on post competency / 中华医学教育探索杂志

Objective:To evaluate the discrimination, reliability and validity of the evaluation system of clinical nurse training based on post competency in hospitals of traditional Chinese and Western medicine.Methods:From August 2018 to January 2019, 271 clinical nurses from a three-A hospital were selected by convenient sampling method. The competency of clinical nurses was assessed on the spot by case tracking method. The discrimination, reliability and validity of the evaluation system were tested by item analysis, Cronbach's α coefficient and confirmatory factor analysis.Results:The evaluation system had good discrimination, reliability and validity. Among the four scales of the evaluation system, there were significant differences in the high score and the low score of all items ( P < 0.05). The Cronbach's α coefficient of each dimension of the four scales was 0.769-0.898. Four structural equation model diagrams were established, the AVE (average variance extracted) of each dimension was 0.51-0.74, factor load was 0.53-0.93, C.R. (composite reliability) was 0.79-0.91, and the discrimination validity was up to the standard. Conclusion:This system provides a reference for the establishment of scientific, objective, measurable and homogeneous clinical nurse training evaluation tools.

A protective effect conferred by KIR3DL1 and its cognate ligand against cervical cancer among ethnic Han Chinese population and its potential mechanism / 中华医学遗传学杂志

Objective@#To explore the role of inhibitory KIR (iKIR) and its cognate HLA ligand in the occurrence and development of cervical cancer among ethnic Han Chinese and its potential mechanism.@*Methods@#Peripheral blood samples from 265 Han Chinese patients with cervical intraepithelial neoplasia (CIN)/cervical cancer and 200 ethnically matched healthy controls were collected. The results of KIR PCR-SSP, HLA PCR-rSSO and KIR3DL1 PCR-SBT, together with cervical cancer data from the TCGA database, were used to assess the association of iKIR genes, receptor-ligand gene combinations, iKIR transcription level in the tumor tissue and the KIR3DL1 alleles with the occurrence and development of cervical cancer.@*Results@#Among the four iKIR genes (KIR2DL1, 2DL2/3, 3DL1 and 3DL2), the frequencies of KIR3DL1 and KIR3DL1-HLA-Bw4 genes among controls were significantly higher than those of the cervical cancer group (96.5% vs. 87.0%, P = 0.018; 81.5% vs. 64.8%, P=0.009). The survival rate of cervical cancer patients with a high transcription level of KIR3DL1 in tumor tissues was significantly higher than those with a low/medium transcription level (P = 0.028). The frequency of strong-inhibitory and high-expression KIR3DL1*01502 allele among the healthy population was significantly higher than that of the cervical cancer group (76.0% vs. 59.3%, P = 0.015).@*Conclusion@#Combined KIR3DL1 and KIR3DL1-HLA-Bw4 can confer a protective effect against the development of cervical cancer, which may be attributed to the strong-inhibitory and high-expression allele of KIR3DL1*01502.

A protective effect conferred by KIR3DL1 and its cognate ligand against cervical cancer among ethnic Han Chinese population and its potential mechanism / 中华医学遗传学杂志

OBJECTIVE@#To explore the role of inhibitory KIR (iKIR) and its cognate HLA ligand in the occurrence and development of cervical cancer among ethnic Han Chinese and its potential mechanism.@*METHODS@#Peripheral blood samples from 265 Han Chinese patients with cervical intraepithelial neoplasia (CIN)/cervical cancer and 200 ethnically matched healthy controls were collected. The results of KIR PCR-SSP, HLA PCR-rSSO and KIR3DL1 PCR-SBT, together with cervical cancer data from the TCGA database, were used to assess the association of iKIR genes, receptor-ligand gene combinations, iKIR transcription level in the tumor tissue and the KIR3DL1 alleles with the occurrence and development of cervical cancer.@*RESULTS@#Among the four iKIR genes (KIR2DL1, 2DL2/3, 3DL1 and 3DL2), the frequencies of KIR3DL1 and KIR3DL1-HLA-Bw4 genes among controls were significantly higher than those of the cervical cancer group (96.5% vs. 87.0%, P = 0.018; 81.5% vs. 64.8%, P=0.009). The survival rate of cervical cancer patients with a high transcription level of KIR3DL1 in tumor tissues was significantly higher than those with a low/medium transcription level (P=0.028). The frequency of strong-inhibitory and high-expression KIR3DL1*01502 allele among the healthy population was significantly higher than that of the cervical cancer group (76.0% vs. 59.3%, P =0.015).@*CONCLUSION@#Combined KIR3DL1 and KIR3DL1-HLA-Bw4 can confer a protective effect against the development of cervical cancer, which may be attributed to the strong-inhibitory and high-expression allele of KIR3DL1*01502.

Construction of competency model for evaluating clinical nurses in hospital of Integrated Traditional Chinese and Western Medicine / 中华医学教育探索杂志

Objective To construct the objective and quantifiable model for evaluating clinical nurses' competency in hospital of Integrated Traditional Chinese and Western Medicine. Methods From December 2016 to August 2017, preliminary formulation of evaluation indicators was constructed through semi-structured interviews, combined with literature analysis. 21 experts from five hospitals of Traditional Chinese Medicine and medical universities were selected using Delphi method for performing two rounds of consultations and weight assignment of indexes. Data was logged and processed using SPSS 16.0. Results The preliminary system of the competency model included four items of Level one and 20 items of Level two. The weight coefficients of the first level indicators were 0.2609, 0.2598, 0.2570 and 0.2224. The positive coefficients from consulations of experts were 100 . 00％ and 95 . 24％, respectively . Authority coefficient was 0.88. The coefficients of variation were 0.0923 to 0.1628 and 0.0430 to 0.1827, respectively. Conclusions This model can provide guidance for the training objective of nurses' competency and serve as an instrument for hospital managers to evaluate nurses in hospital of Integrated Traditional Chinese and Western Medicine.

Construction of competency model for evaluating clinical nurses in hospital of Integrated Traditional Chinese and Western Medicine / 中华医学教育探索杂志

Objective@#To construct the objective and quantifiable model for evaluating clinical nurses' competency in hospital of Integrated Traditional Chinese and Western Medicine.@*Methods@#From December 2016 to August 2017, preliminary formulation of evaluation indicators was constructed through semi-structured interviews, combined with literature analysis. 21 experts from five hospitals of Traditional Chinese Medicine and medical universities were selected using Delphi method for performing two rounds of consultations and weight assignment of indexes. Data was logged and processed using SPSS 16.0.@*Results@#The preliminary system of the competency model included four items of Level one and 20 items of Level two. The weight coefficients of the first level indicators were 0.260 9, 0.259 8, 0.257 0 and 0.222 4. The positive coefficients from consulations of experts were 100.00% and 95.24%, respectively. Authority coefficient was 0.88. The coefficients of variation were 0.092 3 to 0.162 8 and 0.043 0 to 0.182 7, respectively.@*Conclusions@#This model can provide guidance for the training objective of nurses' competency and serve as an instrument for hospital managers to evaluate nurses in hospital of Integrated Traditional Chinese and Western Medicine.

Discussion on the Issues and Countermeasures in Clinical Trial Insurance / 中国医学伦理学

Through analyzing the current situation and coverage controversy of Chinese clinical trial insurance,this paper stated that the attending insurance rate in domestic clinical trials was entirely low.The sponsors,clinical trial institutions,investigators and insurance companies paid attention of different levels to clinical trial insurance.Therefore,the risk awareness of drug/medical device clinical trials should be enhanced.It is necessary to give impetus to clinical trial insurance system,during which all parties need to make a joint effort including government departments,ethics committees,sponsors,clinical trial institutions,investigators and insurance companies.

Carcino-embryonic antigen targeted and drug loaded ultrasound nanoparticle agents inhibit growth of ovarian cancer cells in vitro / 中国医学影像技术

Objective To prepare carcino-embryonic antigen (CEA) targeted and paclitaxel loaded phase-shifting PLGA nanoparticles (Ab-PTX-NPs),and investigate the targeting capability and inhibition to the ovarian cancer cell in vitro.Methods Single-emulsion/solvent evaporation (O/W) and carbodiimide method were used to prepare the Ab-PTX-NPs.The size of nanoparticles was determined by Malvern analyzer.The encapsulation and drug loaded efficiency of paclitaxel were detected by high performance liquid chromatography.And the drug release characteristics was measured by dialysis method in constant temperature shaker.The targeting ability of Ab-PTX-NPs to the ovarian cancer SKOV3 cell was evaluated by the laser scanning confocal microscope and flow cytometry.And the inhibition ability of Ab-NPs was investigated by the CCK-8 assays.Results The size of Ab-PTX-NPs was (397.70±99.95)nm.The encapsulation efficiency and drug loading capacity of PTX were (67.26±4.15) ％ and (6.31±0.39) ％,respectively.The conjugating rate of Anti-CEA antibody was (92.74 ± 5.75) ％.The targeting study in vitro showed that such a number of contrast agents landed around the SKOV3 cells in targeting group,and the mean fluorescence intensity of ovarian cells in targeting group was significantly higher than other groups (P＜0.05).After 24 h,the viability rate of ovarian cells in targeting group was lower than the non-target group (P＜0.05),only higher than that of the pure PTX group (P＜0.05).But there was no significant difference between the targeting group and the pure PTX group (P＞0.05) at 48 h.Conclusion The CEA targeted and paclitaxel loaded phase-shifting PLGA nanoparticles are successfully prepared.It can enhance ultrasound imaging well after activated by LIFU.With high drug-loading efficiency and fast drug release velocity,the Ab-PTX-NPs appeares great targeted ability.

Targeted multifunctional nanoparticles probe mediated photoacoustic imaging and treatment study of breast carcinoma in vitro / 中华超声影像学杂志

Objective To investigate the ability of targeting in vitro breast carcinoma and photoacoustic-mediate apoptosis of breast carcinoma of the nanoparticle probe loaded with Fe3O4 and observe its consequence of photoacoustic imaging in vitro.Methods The polymeric multifunctional nanoparticles probe that was loaded with Fe3O4 and connected with Herceptin targeting breast carcinoma was prepared in the use of double emulsion method and carbodiimide method.General physical property,the condition of Herceptin connected with nanoparticles and the ability of targeting of the probe were tested.The different concentration of nanoparticles was imaged by photoacoustic.The inhibiting effect of targeting nanoparticles on breast carcinoma cells was evaluated in vitro.Results The targeting polymeric multifunctional nanoparticles probes loaded with Fe3O,were prepared successfully with average particle diameter of (235.4± 53.75)nm and Zeta potential (-13.4 ± 4.7)mV.Fe3O4 particles dispersed on the shell of the probes.Antibody Hereeptin was successfully connected with the surface of the nanoprobes.There were massive targeting nanoprobes surround the breast carcinoma cell strains SKBR3 in the targeting group in vitro.The photoascoustic signal of the nanoprobes enhanced with the increase of Fe3O4 concentration in photoacoustic experiment in vitro.The apoptotic rate of breast carcinoma increased after the laser irradiation in the cell inhibition experiment in vitro.That proved the obvious inhibition of breast carcinoma cells was caused by photothermal effect of the prepared nanoprobes.Conclusions The prepared polymeric multifunctional nanoparticles probe that was loaded with Fe3O4 and connected with Herceptin targeting breast carcinoma can be used as a photoacoustic contrast agent,which can inhibit the proliferation of breast carcinoma by targeting photoacoustic therapy.

Preparation and characteristics of targeted phase-shift lipid nanoparticles mediated by tumor homing and penetrating peptide / 中华超声影像学杂志

Objective To prepare a novel ultrasound contrast agent , targeted phase-shift lipid nanoparticles mediated by tumor homing and penetrating peptide tLyP-1 ,and to evaluate its characteristics . Methods The nanoparticles were prepared by filming-rehydration and acoustic-vibration methods .The morphology ,distribution ,particle size and zeta potential were detected . After heating and irradiating of low intensity focused ultrasound ( LIFU) ,the phase-shift characteristic and the enhancement effect in vitro were observed . The tumor homing and cell-penetrating properties of the nanoparticles were examined by confocal laser scanning microscopy and flow cytometry . The cytotoxicity of the nanoparticles was evaluated by CCK 8 assay . Results The size and distribution of nanoparticles were uniformed . The size and zeta potential of nanoparticles were ( 399 .50 ± 29 .98) nm and ( 3 .28 ± 1 .72) mV ,respectively . When the nanoparticles were heated to a temperature of 45 ℃ or after irradiated by LIFU ,nanoparticles generated phase-shift and enhanced ultrasound imaging in vitro ( P 0 .05 ) . Conclusions A novel ultrasound contrast agent , targeted phase-shift lipid nanoparticles mediated by tumor homing and penetrating peptide tLyP-1 ,is prepared successfully . It can target to MDA-MB-231 cell and penetrate into the cell in vitro ,and enhance ultrasound imaging in vitro after LIFU irradiation ,which expected to become a novel tumor targeted ultrasound contrast agent and achieve ultrasound molecular imaging at the level of tumor cell .

Research progress of application of traditional Chinese medicine nursing in cervical spondylosis / 中国实用护理杂志

The effects of Traditional Chinese medicine (TCM) nursing operation in cervical spondylitis have been generally recognized. This paper reviewed the applications and improvements of the major three TCM nursing operations, cupping, Gua-Sha and acupressure. The future developments in moxibustion, auricular point sticking, fumigation and washing therapy and moisten compress therapy were also prospected.

Bioequivalence study on two terazosin hydrochloride tablets / 药学实践杂志

Objective To evaluate the bioequivalence of domestic and imported terazosin hydrochloride tablets after single oral dose .Methods It was a single center ,randomized ,open ,cross-over trail design ,21 subjects were fasting oral adminis-tered of 2 mg domestic and imported terazosin hydrochloride tablets in different periods ,venous blood 4 ml were collected in different time points before and 60 h after administration ,plasma concentration of terazosin was determined by LC-MS/MS . Results The main pharmacokinetic parameters of domestic and imported terazosin hydrochloride tablets were as follows :t1/2 :(13.2± 2.39)hvs(12.5±1.93)h,tmax :(1.01±0.83)hvs(1.08±0.69)h,Cmax :(40.1±10.6)ng/mlvs(37.3± 9 .57) ng/ml;AUC0- ∞ :(428 ± 82 .1) ng · h/ml vs (426 ± 85 .2) ng · h/ml .The relative bioavailability of domestic terazosin hydrochloride tablets was (101 .2 ± 14 .7)% .90% CI of domestic and imported terazosin hydrochloride tablets AUC0-t and Cmax geometric mean ratio fell between 80% -125% .Conclusion The domestic tablets are bioequivalent to the imported tablets .

Some Thoughts on Clinical Efficacy Evaluation of Traditional Chinese Medicine / 世界科学技术-中医药现代化

The research on clinical efficacy evaluation of traditional Chinese medicine (TCM) is a hot and difficult issue in the field of TCM. However, the overall level of current related research also needs to be improved. There are still a lot of academic controversies in the field of TCM clinical efficacy evaluation. The author believed that before conducting research, it is necessary to sort out core issues of TCM clinical efficacy evaluation, which in-clude the concept of TCM clinical efficacy and its evaluation. In recent years, Chinese scholars have conducted a series of studies on these issues and have some achievements. To think and answer these questions will play an important role for the ultimate resolution of problem of TCM clinical efficacy evaluation.

An evaluation method for analysis of correlation between traditional Chinese medicine syndrome and seasonal changes of weather based on information entropy / 中西医结合学报

OBJECTIVE: A method based on dubious condition of information entropy was introduced and applied to discuss a complexity problem in the analysis of correlation between traditional Chinese medicine (TCM) syndrome and season. METHODS: Eight hundred and seventy one cases of chronic virus hepatitis B (hepatitis B) with TCM clinical data were analyzed by information entropy method. RESULTS: It was found that hepatitis B with Yin deficiency of liver and kidney happened more often in summer than in other seasons. CONCLUSION: It is inferred that the difference of seasons may influence the variation of TCM syndromes.

Professor JIANG Jian's clinical experience in treating pain syndrome with later generation's prescriptions / 中华中医药杂志

This article introduced five cases of professor JIANG Jian using later generation's prescriptions of Qingwei Shegan Tang,Qingkong Gao,Xietong Shen Fang,Jianbei Jingutong Fang,Juantong San,Juhe Wan,and Shen xiao Gualou San to treat pain syndrome of gastralgia,hypochondriac pain,headache,backache,and breast-ache. During the treatment of pain syndrome,professor JIANG Jian’ clinical experience by using later generation's prescriptions was extraordinary,deserved to learn an reference.