Establishment of CNC regional microbial database and microbial population analysis for pharmaceutical enterprises / 中国生物制品学杂志

@#Objective To establish CNC(controlled not classified)area microbial database of pharmaceutical enterprises,conduct correlation analysis with microbial distribution in clean area to achieve good microbial control in the clean area,especially in the core aseptic operation area,and improve the sterility assurance level of pharmaceutical enterprises.MethodsThe CNC area of a manufacturing workshop from pharmaceutical enterprise was selected as the research object,the airborne microbe,settling microbe,personal and environmental surface microorganisms were sampled one by one in the CNC area according to the route of personnel and materials entering the clean area. After the microorganisms were cultured and purified,the 16S rRNA sequencing method was used to identify the strains. Microbiomeanalyst online analysis software was used for data analysis.ResultsThe number of microorganisms in the CNC area of pharmaceutical enterprises was large and the species were rich. A total of 4 080 microorganisms were collected,which distributed in 47 genera. Gram-positive bacteria accounted for 84. 4%,mainly belonging to Staphylococcus,Micrococcus,Microbacterium. Gram-negative bacteria accounted for 15. 6%,and the distribution of Acinetobacter was much higher than other gram-negative bacteria. The Alpha diversity index was used to characterize the richness of microbial populations. The richness of microbial populations of airborne microbe and settling microbe was the highest,followed by environmental surface microorganisms,and the microbial population richness of personnel surface microorganism was the lowest. The number of microorganisms in the personnel's hood and chest was relatively small,and the number of microorganisms in the personnel's soles of feet and hands was relatively large. There were relatively few types of microorganisms on the surface of personnel,and staphylococcus showed an absolute advantage. The microbial distribution in CNC area was correlated with the microbial distribution in Class B and C clean areas of the same production workshop. Most of the microorganisms collected in Class B and C clean areas were included in the microbial population in CNC area. Class B and C clean areas were similar to CNC area,both of which were dominated by Staphylococcus and Micrococcus. The distribution proportion of Acinetobacter in Class B and C clean areas decreased compared with that in CNC area.ConclusionThe CNC area microbial database of the pharmaceutical enterprises has been established and the microbial population that is easy to enter the clean area has been determined,providing reliable technical support for the good microorganism control in the clean area

Comparison on the clinical characteristics of patients with Takotsubo syndrome from China and from Europe/North America / 中华心血管病杂志

Objective: To summarize the clinical characteristics of patients with Takotsubo syndrome (TTS) from China and compare these features with patients from Europe/North America. Methods: We reviewed case reports published between 1990 and 2020 with the key words of "Takotsubo syndrome" "stress cardiomyopathy" "apical balloon syndrome" and "broken heart syndrome", in Wanfang, CNKI, Pubmed and Web of Science databases, and 1 294 articles were identified, including 128 articles reporting 163 cases in China and 1 166 articles reporting 1 256 cases in Europe/North America. The characteristics of demographics, triggers, symptoms, electrocardiogram, echocardiography, left ventriculogram,coronary angiography, treatment and prognosis were analyzed and compared between Chinese and European/North American cases. Results: A total of 1 294 articles (1 419 cases: 163 from China, 1 256 from Europe/North America) were included in the final analysis. The characteristics of Chinese cases included: (1) demographic:the age was (59.6±16.9) years, which was similar with that of European/North American ((59.7±17.4) years, P=0.90), and female accounting for 78.5% (128/163), which was lower than that of European/North American (85.4% (1 073/1 256), P=0.02). (2) Triggers:mental triggers accounted for 48.5% (79/163), physical triggers accounted for 43.6% (71/163), and no triggers accounted for 7.9% (13/163), respectively. Compared with Europe/North America, the ratio of patients with mental triggers was higher in China, while the ratio of patients with physical triggers and no triggers was lower (P<0.05). (3) Symptoms: chest pain (52.8% (86/163)), chest tightness (35.0% (57/163)), shortness of breath (33.1% (54/163)), dizziness (16.0% (26/163)), sweating (15.3% (25/163)), palpitations (12.3% (20/163)), syncope (9.2% (15/163)) abdominal pain/diarrhea (8.6% (14/163)), hypotension (7.4% (12/163)), and fatigue (1.2% (2/163)) were illustrated in sequence. Compared with patients in Europe/North America, the ratio of patients with chest tightness, dizziness, sweating, palpitations, abdominal pain/diarrhea was higher in Chinese patients, while the ratio of patients with hypotension was lower in Chinese patients (P<0.05). (4) Electrocardiogram: main manifestations were myocardial ischemia symptoms, such as ST-segment elevation (63.8% (104/163)), T wave inversion (46.0% (75/163)), ST-segment depression (8.6% (14/163)). Compared with European/North American, the ratio of patients with ST-segment elevation, T wave inversion, and atrioventricular block was higher in Chinese patients (P<0.05). (5) Echocardiography and imaging:apical dyskinesia (59.5% (97/163)) and apical/left ventricular bulbar dilation (36.2%(59/163)) dominated the echocardiography findings. Compared with European/North American, the ratio of patients with apical dyskinesia, apical/left ventricular bulbar dilation, and mitral regurgitation was higher in Chinese patients, while the ratio of patients with dyskinesia in other parts and left ventricular ejection fraction<50% was lower in Chinese patients (P<0.05). Left ventricular angiography showed 36.2% (59/163) of apical dyskinesia in Chinese patients, which was higher than that reported in European/North American patients, and 38.7% (63/163) of apical/left ventricular bulbar dilation was reported in Chinese patients, which was similar to that reported in European/North American patients. Coronary angiography showed percent of no stenosis or stenosis less than 50% was 87.1% (142/163), which was similar to that reported in European/North American patients (P>0.05). The typical type of TTS accounted for 96.3% (157/163), which was significantly higher than that reported in European/ American patients, while the ratio of basal type and midventricular type was lower (P<0.01). (6) Treatment and prognosis:the applied drugs in China were listed in order as following, β-blockers (41.1% (67/163)), antiplatelet agents (37.4%(61/163)), ACEI/ARB (36.2%(59/163)), anticoagulants (27.0%(44/163)), diuretics (19.6% (32/163)), etc. Compared with Europe/North America, the ratio of antiplatelet agents, anticoagulants, statins, diuretics, and nitrates use was higher in China (P<0.05), while the use of oxygen therapy and IABP was similar (P>0.05). The hospital mortality in China was 5.5% (9/163), during 1-year follow-up the recurrence rate was 3.7% (6/163) and the mortality was 0. The prognosis was similar with that in Europe/North America. Conclusions: Compared with TTS cases in Europe/North America, TTS cases in China also occur usually in middle-aged and elderly women, most of whom have mental/physical triggers and typical imaging manifestations, followed by a low hospital mortality rate and recurrence rate.

Effect of intervention on high caries risk children in some kindergartens in Zhangjiagang / 中国学校卫生

Objective@#To explore the prevention and control effects of different management modes on children with high risk of dental caries from some kindergartens in Zhangjiagang, and to provide a basis for relevant departments to formulate oral health management strategy.@*Methods@#In September 2020, 1 600 children aged 3 years old from 9 towns in Zhangjiagang were sampled by cluster sampling method for baseline survey, including oral examination, questionnaire survey and caries susceptibility test. According to the risk assessment criteria, high risk children were screened out and divided into two groups. In the experimental group, the frequency of fluoride application was once every 3 months, combined with behavioral management and oral health guidance. In the control group, fluoride application was conducted once every 6 months, combined with behavioral management and oral health guidance. The effect of caries prevention was evaluated one year later.@*Results@#The prevalence of caries in three year old children was 50.9% at baseline, and the prevalence of caries in boys and girls was 47.5% and 54.5%, respectively, with statistical significance ( χ 2=9.64, P <0.05). A total of 1 090 high risk children were screened out, including 475 in experimental group and 615 in control group. The prevalence of caries in the two groups at baseline was 74.1% and 75.1%, respectively, and dmft were (2.98±3.33) and (3.04±3.16), respectively, with no significant difference ( χ 2/t =0.15, 0.28, P >0.05). One year after intervention, the prevalence of caries in the two groups was 78.5% and 83.0%, respectively and dmft were (4.22±3.97) and (4.51±4.08), respectively, with no statistical significance ( χ 2/t=3.17, 0.05, P >0.05). The incidence of new caries in the experimental group was 3.7%, and the incidence of new caries was (1.26±1.69), lower than 7.6% and (1.45±2.04) in the control group, with statistical significance ( χ 2/t=6.89, 5.05, P <0.05).@*Conclusion@#The frequency of fluorination intervention once every 3 months combined with behavior management mode was more effective in controlling new caries in children with high risk of caries.

Comparison of Relative Molecular Weight Distribution and Immune Activity of Polysaccharides in Polygonati Rhizoma Before and After Processing / 中国实验方剂学杂志

Objective:To study the relative molecular weight and distribution of polysaccharides in Polygonati Rhizoma （PR） before and after processing， as well as the effects of different polysaccharide fractions on immune function and inflammatory response of mouse peritoneal macrophages. Method:High-performance gel permeation chromatography （HPGPC） was used to determine the relative molecular weight and distribution of polysaccharides in PR （named SC） and polysaccharides in PR processed with wine （named JC）， and polysaccharide fractions with different relative molecular weights were obtained by dialysis. Different polysaccharide fractions were applied to mouse peritoneal macrophages， which was normal or induced by lipopolysaccharide （LPS）. Cell counting kit-8 （CCK-8） method was used to select the optimal administration concentration. Enzyme linked immunosorbent assay （ELISA） was used to determine the concentrations of tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） and interleukin-1<italic>β</italic> （IL-1<italic>β</italic>） in the cell supernatant. The Griess method was used to detect the concentration of nitric oxide （NO）. Result:SC and JC could be divided into four parts according to relative molecular weight and its distribution range， including part Ⅰ（14 800-2 273 kDa）， part Ⅱ（2 148-296 kDa）， part Ⅲ（12-1 kDa） and part Ⅳ（818-362 Da）. Based on the differences of part Ⅰ and part Ⅲ after processing， the SC and JC were divided into two-part according to the weight-average relative molecular weight （<italic>M</italic>_W）. For normal mouse peritoneal macrophages， JC could significantly promote the secretion of TNF-<italic>α</italic> （<italic>P</italic><0.01）， while SC had no significant effect. Four polysaccharide fractions， named SD （SC fraction with <italic>M</italic>_W>50 kDa）， JD （JC fraction with <italic>M</italic>_W>50 kDa）， SX （SC fraction with <italic>M</italic>_W<50 kDa） and JX （JC fraction with <italic>M</italic>_W<50 kDa）， also could<italic> </italic>significantly promote the secretion of TNF-<italic>α</italic> （<italic>P</italic><0.01）， but only JX could significantly promote the secretion of NO （<italic>P</italic><0.05）. In addition， the effect of JX group stimulated secretion of TNF-<italic>α</italic> was better than the JD group （<italic>P</italic><0.01）. For the LPS-induced macrophage model， JC and SC group could<italic> </italic>significantly inhibit the secretion of TNF-<italic>α</italic> and IL-1<italic>β</italic> （<italic>P</italic><0.01）， and the effect of JC was stronger. To compare different polysaccharide fractions， the impact of JX on inhibiting the secretion of TNF-<italic>α</italic> and IL-1<italic>β</italic> was<italic> </italic>significantly stronger than JD （<italic>P</italic><0.01）， and SX inhibited the secretion of TNF-<italic>α</italic> was significantly stronger than SD （<italic>P</italic><0.01）. Conclusion:The relative molecular weight and distribution of polysaccharides in PR before and after processing have changed. JC and SC improve the immune regulation mainly by inhibiting the inflammatory response， the fraction of <italic>M</italic>_W<50 kDa is the main effective part， and the effect of PR polysaccharides in inhibiting the inflammatory is enhanced after processing with wine.

Protective effect of CD73 inhibitor α, ß-methylene ADP against amyloid-ß-induced cognitive impairment by inhibiting adenosine production in hippocampus

BACKGROUND: Alzheimer's disease (AD) is a chronic, progressive neurodegenerative disease. Recent studies have reported the close association between cognitive function in AD and purinergic receptors in the central nervous system. In the current study, we investigated the effect of CD73 inhibitor α, ß-methylene ADP (APCP) on cognitive impairment of AD in mice, and to explore the potential underlying mechanisms. RESULTS: We found that acute administration of Aß1­42 (i.c.v.) resulted in a significant increase in adenosine release by using microdialysis study. Chronic administration of APCP (10, 30 mg/kg) for 20 d obviously mitigated the spatial working memory impairment of Aß1­42-treated mice in both Morris water maze (MWM) test and Y-maze test. In addition, the extracellular adenosine production in the hippocampus was inhibited by APCP in Aß-treated mice. Further analyses indicated expression of acetyltransferase (ChAT) in hippocampus of mice of was significantly reduced, while acetylcholinesterase (AChE) expression increased, which compared to model group. We observed that APCP did not significantly alter the NLRP3 inflammasome activity in hippocampus, indicating that anti-central inflammation seems not to be involved in APCP effect. CONCLUSIONS: In conclusion, we report for the first time that inhibition of CD73 by APCP was able to protect against memory loss induced by Aß1­42 in mice, which may be due to the decrease of CD73-driven adenosine production in hippocampus. Enhancement of central cholinergic function of the central nervous system may also be involved in the effects of APCP.

Successful treatment of a patient with cutaneous co-infection caused by Mucor irregularis and Klebsiella pneumoniae

Abstract The authors report a rare case of primary cutaneous mucormycosis caused by Mucor irregularis and cutaneous Klebsiella pneumoniae infections in a 67-year-old Chinese woman. After the administration of liposomal amphotericin B combined with cefoperazone/sulbactam sodium, the patient recovered. Invasive fungal infection combined with cutaneous bacterial infection should receive attention.

Research on Textual Research and Methods of Chinese Herbs in Famous Classical Formulas / 中国实验方剂学杂志

As the essential part of tratidional Chinese medicine(TCM), the research and development of classic formula have become a hot spot in TCM industry. However, with the change of the age, the species, medical part and origin of TCM have more or less changed. It is of great significance for the safety and effectiveness of the classical prescription to clarify the varieties and medicinal parts of TCM. In this paper, based on the discussion of the methods of textual research on the Chinese herbs, the species and medical parts, origin of Chinese herbs in a list of 100 famous classical formulas which promulgated by the state administration of TCM were analyzed. The textual research of Chinese herbs shows that most of the herbs involved in the classical formula have the problems of species, medical part, and origin. Therefore, it is of great significance for the selection of the species and medical parts, origin of the Chinese herbs in the research and development process of the classical formula.

Correlation between long non-coding RNAs (lncRNAs) H19 expression and trastuzumab resistance in breast cancer

Introduction: Trastuzumab resistance is a major obstacle encountered in human epidermal growth factor receptor 2 (HER2)-positive breast cancer therapy. Long non-coding RNAs (lncRNAs) have been confirmed to play important roles in both tumorigenesis and tumor development. However, whether lncRNAs are associated with trastuzumab resistance is not yet clear. Subjects and Methods: We evaluated trastuzumab sensitivity in breast cancer cell lines, SKBR3, HCC1954, and MDA-MB-231. We also evaluated H19 expression in these cell lines after treatment with different trastuzumab concentrations. Besides, H19 was downregulated to investigate its role in cell viability and trastuzumab sensitivity and a trastuzumab resistance cell line was cultured to verify the effect of H19 in trastuzumab resistance. Forty-eight HER2-positive breast cancer patients treated with trastuzumab in the first-line setting were selected retrospectively to explore the relationship between H19 expression and tumor-node-metastasis (TNM) stage as well as trastuzumab resistance. Results: H19 is a trastuzumab-responsive lncRNA and its expression was upregulated in a trastuzumab-resistant breast cancer cell. Downregulation of H19 restored the sensitivity of trastuzumab-resistant cells to this drug. The expression of H19 significantly correlated with TNM stage. Patients with higher expression of H19 showed an evidently shorter progression-free survival than those with low H19 expression. H19 overexpression was negatively correlated to the trastuzumab-therapy response. Conclusions: Our results provide evidence for the H19-mediated regulation of trastuzumab resistance in HER2-positive breast cancer cells. H19 could act as a potential predictive biomarker for HER2-positive breast cancer patients, and downregulation of H19 could reverse trastuzumab resistance and enhance the inhibitory function of this drug

Comparison of Total Daily Doses of 16 Active Components in Three Dosage Forms of Fuzi Lizhongwan / 中国实验方剂学杂志

Objective:To compare the total daily doses of 16 active components in big honeyed pills, concentrated pills and tablets of Fuzi Lizhongwan. Method:Three dosage forms of Fuzi Lizhongwan were prepared according to the process described in the literature. RRLC-QqQ-MS was employed to analyze the contents of 16 active ingredients with mobile phase of 0.1%formic acid aqueous solution-0.1%formic acid acetonitrile solution for gradient elution,the separation was performed on a Accucore RP-MS column(2.1 mm×100 mm, 2.6 μm) with a flow rate of 0.3 mL·min-1 and the column temperature at 30℃, the mass spectrometry condition was electrospray ion source, positive and negative ion switching mode for detection, multi-reaction monitoring mode(MRM) for scanning. The contents of 16 active ingredients were calculated, and the normalization arithmetic method was used for comparing the total daily doses of these active ingredients in three dosage forms of Fuzi Lizhongwan. Result:Processed products of Aconiti Lateralis Radix Praeparata were used as raw powder in preparation process of the three dosage forms, so there was no significant difference in the contents of six alkaloids in the three dosage forms, while the contents of other 10 active ingredients from Zingiberis Rhizoma, Codonopsis Radix, Atractylodis Macrocephalae Rhizoma and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were significantly higher in big honeyed pills than those in concentrated pills or tablets(PConclusion:The total daily doses of 16 active ingredients in the three dosage forms of Fuzi Lizhongwan are significantly different caused by preparation process, prescription and dosage.

Arsenic trioxide induces cell cycle arrest and affects Trk receptor expression in human neuroblastoma SK-N-SH cells

BACKGROUND: Arsenic trioxide (As2O3), a drug that has been used in China for approximately two thousand years, induces cell death in a variety of cancer cell types, including neuroblastoma (NB). The tyrosine kinase receptor (Trk) family comprises three members, namely TrkA, TrkB and TrkC. Various studies have confirmed that TrkA and TrkC expression is associated with a good prognosis in NB, while TrkB overexpression can lead to tumor cell growth and invasive metastasis. Previous studies have shown that As2O3 can inhibit the growth and proliferation of a human NB cell line and can also affect the N-Myc mRNA expression. It remains unclear whether As2O3 regulates Trks for the purposes of treating NB. METHODS: The aim of the present study was to investigate the effect of As2O3 on Trk expression in NB cell lines and its potential therapeutic efficacy. SK-N-SH cells were grown with increasing doses of As2O3 at different time points. We cultured SK-N-SH cells, which were treated with increasing doses of As2O3 at different time points. Trk expression in the NB samples was quantified by immunohistochemistry, and the cell cycle was analyzed by flow cytometry. TrkA, TrkB and TrkC mRNA expression was evaluated by real-time PCR analysis. RESULTS: Immunohistochemical and real-time PCR analyses indicated that TrkA and TrkC were over-expressed in NB, and specifically during stages 1, 2 and 4S of the disease progression. TrkB expression was increased in stage 3 and 4 NB. As2O3significantly arrested SK-N-SH cells in the G2/M phase. In addition, TrkA, TrkB and TrkC expression levels were significantly upregulated by higher concentrations of As2O3 treatment, notably in the 48-h treatment period. Our findings suggested that to achieve the maximum effect and appropriate regulation of Trk expression in NB stages 1, 2 and 4S, As2O3 treatment should be at relatively higher concentrations for longer delivery times;however, for NB stages 3 and 4, an appropriate concentration and infusion time for As2O3 must be carefully determined. CONCLUSION: The present findings suggested that As2O3 induced Trk expression in SK-N-SH cells to varying degrees and may be a promising adjuvant to current treatments for NB due to its apoptotic effects.

Inhibition of Adhesion and Metastasis of HepG2 Hepatocellular Carcinoma Cells In Vitro by DNA Aptamer against Sialyl Lewis X / 华中科技大学学报（医学）（英德文版）

The sialyl Lewis X (SLex) antigen encoded by the FUT7 gene is the ligand of endotheliam-selectin (E-selectin).The combination of SLex antigen and E-selectin represents an important way for malignant tumor metastasis.In the present study,the effect of the SLeX-binding DNA aptamer on the adhesion and metastasis of hepatocellular carcinoma HepG2 cells in vitro was investigated.Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining were conducted to detect the expression of FUT7 at both transcriptional and translational levels.The SLex expression in HepG2 cells treated with different concentrations of SLeX-binding DNA aptamer was detected by flow cytometry.Besides,the adhesion,migration,and invasion of HepG2 cells were measured by cell adhesion assay,and the Transwell migration and invasion assay.The results showed that the FUT7 expression was up-regulated at both mRNA and protein levels in HepG2 cells.SLeX-binding DNA aptamer could significantly decrease the expression of SLex in HepG2 cells.The cell adhesion assay revealed that the SLeX-binding DNA aptamer could effectively inhibit the interactions between E-selectin and SLex in the HepG2 cells.Additionally,SLeX-binding DNA aptamers at 20 nmol/L were found to have the similar effect to the monoclonal antibody CSLEX-1.The Transwell migration and invasion assay revealed that the number of penetrating cells on the down-side of Transwell membrane was significantly less in cells treated with 5,10,20 nmol/L SLeX-binding DNA aptamer than those in the negative control group (P＜0.01).Our study demonstrated that the SLeX-binding DNA aptamer could significantly inhibit the in vitro adhesion,migration,and invasion of HepG2 cells,suggesting that the SLeX-binding DNA aptamer may be used as a potential molecular targeted drug against metastatic hepatocellular carcinoma.

Knockdown of GRHL3 Inhibits Activities and Induces Cell Cycle Arrest and Apoptosis of Human Colorectal Cancer Cells / 华中科技大学学报（医学）（英德文版）

The Grainyhead-like 3 (GRHL3) is involved in epidermal barrier formation,neural tube closure and wound repair.Previous studies have suggested that GRHL3 has been linked to many different types of cancers.However,to date,its effects on human colorectal cancer (CRC) has not been clarified yet.Our microarray analysis has indicated predominant GRHL3 expression in CRC.The purpose of this study was to investigate the expression and significance of GRHL3 in CRC tumorigenesis using CRC tissues and paired paracancerous tissues,as well as using distinct CRC cell lines (HT29 and DLD1).We observed increased GRHL3 expression at both mRNA and protein levels in CRC tissues and CRC cell lines using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting.Moreover,silencing GRHL3 with siRNA could suppress CRC cell proliferation,viability and migration in vitro.We also found that knockdown of GRHL3 could promote cell cycle arrest at G0/G1 phase in HT29 cells and DLD1 cells,and induce cell apoptosis in HT29 cells.Together,our study revealed the down-regulation of GRHL3 in vitro could inhibit CRC cell activity and trigger cell cycle arrest at G0/G1 phase and apoptosis.

Study on HPLC fingerprint and chemical constituent difference of different species of Aurantii Fructus Immaturus / 中国中药杂志

This study is to establish an HPLC fingerprint by HPLC-DAD method and simultaneous quantitative analysis of 17 components of 18 batches of Citrus aurantium and 10 batches of C. sinensis. The separation was performed on an Agilent Poroshell 120 SB-C₁₈ (4.6 mm×100 mm,2.7 μm) column with the gradient elution of methanol-0.1% formic acid water, the flow was 0.6 mL•min⁻¹. The detection wavelength was set at 318 nm. The column temperature was maintained at 30 ℃. The data calculation was performed with similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (Version 2004A) together with SIMCA-P 13.0 software to clarify the differential marker between these two different species of Aurantii Fructus Immaturus. This method has good precision stability and repeatability that could provide basis for quality control and evaluation of Aurantii Fructus Immaturus.

Research on concentration of 5 different ginsenosides in Panax japonica collected from different cultivated geographic regions / 中国中药杂志

In this paper, an HPLC-QqQ-MS method for determination of 5 different ginsenosides of Panax japonica collected from different cultivated geographic regions was established. The separation was performed on a Zorbax XDB-C₁₈ (4.6 mm×100 mm, 1.8 μm) column with the gradient elution of acetonitrile (contained 0.1% formic acid)-0.1% formic acid water. The flow rate was 0.5 mL•min⁻¹. The colunm temperature was maintained at 30 ℃. The analytes were detected using electrospray ionization (ESI) in multiple reaction monitoring (MRM) modes. Reaction selected ions were 203.2 for ginsenoside Re, 202.9 for ginsenoside Rg₁, 365.0 for ginsenoside Rf, 789.1 for ginsenoside Rd, 360.9 for ginsenoside Ro. Ginsenosides Re, ginsenosides Rg₁, ginsenosides Rf, ginsenosides Rd, ginsenosides Ro had good linearity in the ranges of 3.33-66.60 μg (r=0.999 1),2.83-56.54 μg (r=0.999 2), 0.32-6.51 μg (r=0.999 2), 12.55-251.00 μg (r=0.999 3), 0.85-16.90 μg (r=0.999 5), respectively. The results of recovery were among 100.8% to 104.6%, and the values of RSD were blow 3.0%. This method is simple, reliable and accurate, and can provide basis for P. japonica basic research.

Suppression of Allogeneic T Cells Proliferation by CD3/CD46-Induced T-regulatory 1 Cells / 华中科技大学学报（医学）（英德文版）

CD46 is not only identified as a complement regulatory protein which protects host cells from complement attack,but also a new co-stimulatory molecule for human T cells.CD3/CD46 co-stimulation can induce a T-regulatory 1 cell(Tr1)-specific cytokine phenotype in human CD4+T cells.However,the role of CD46 as a co-stimulatory molecule in the modulation of the acquired immunity,such as transplant immunology,remains unclear.In this study,CD4+T cells were isolated from human CD46-transgenic C57BL/6 mice by magnetic-activated cell sorting,and further induced by anti-CD3,anti-CD28 and anti-CD46 antibodies respectively,and anti-CD3/anti-CD28 antibodies,anti-CD3/anti-CD46 antibodies,or the monoclonal antibody panel against CD3/CD28/CD46.The levels of interleukin-2(IL-2),γ-interferon(γ-IFN),interleukin-10(IL-10)and transforming growth factor-β(TGF-β)were detected in the supematants of different groups.Suppression of allogeneic T cell proliferation were assessed by using mixed lymphocyte reaction(MLR)assay,in which monoclonal antibodies against CD46 were added to the culture.The results showed that CD3/CD28,CD3/CD46 and CD3/CD28/CD46 co-stimulation could significantly induce stronger proliferation of T cells than CD3 stimulation(P＜0.05),and CD3/CD28/CD46 co-stimulation significantly increased the proliferation of T cells when compared with CD3/CD28 or CD3/CD46 co-stimulation(P＜0.05 for each).IL-2 and γ-IFN levels were much higher in CD3/CD28 co-stimulation group than in CD3,CD28,CD46 and CD3/CD46 groups(P＜0.05 for each).IL-10 and TGF-β levels were dramatically increased in CD3/CD46 co-stimulation group as compared with those in the CD3,CD28,CD46 and CD3/CD28 groups(P＜0.05 for each).CD3/CD46 co-stimulation significantly inhibited the T cell proliferation and allogenic immune responses through the secretion of IL-10 and TGF-β in MLR (P＜0.05).These results suggested that CD3/CD46 can induce Tr1 cells to modulate allogenic immune responses,and it may become a novel target for the development of new therapeutic approach for T-cell-mediated diseases.CD46 plays an important role in regulating the T cell-mediated immune responses by bridging innate and acquired immunity.