Analysis of Alterations in Intestinal Flora in Chinese Elderly with Cardiovascular Disease and Its Association with Trimethylamine.

To investigate the changes in the intestinal flora in the Chinese elderly with cardiovascular disease (CVD) and its correlation with the metabolism of trimethylamine (TMA), the intestinal flora composition of elderly individuals with CVD and healthy elderly individuals was analyzed using 16S rRNA sequencing, the TMA levels in the feces of elderly were detected using headspace-gas chromatography (HS-GC), and four kinds of characterized TMA-producing intestinal bacteria in the elderly were quantified using real-time fluorescence quantitative polymerase chain reaction (qPCR). The results showed that Firmicutes, Actinobacteria, Proteobacteria, Bacteroidetes, and Verrucomicrobia are the dominant microorganisms of the intestinal flora in the Chinese elderly. And there were significant differences in the intestinal bacteria composition between healthy elderly individuals and those with CVD, accompanied by a notable difference in the TMA content. The richness and diversity of the intestinal flora in the elderly with CVD were higher than those in the healthy elderly. Correlation analysis indicated that certain significantly different intestinal flora were associated with the TMA levels. Our findings showed a significant difference in TMA-producing intestinal flora between healthy elderly individuals and those with CVD. The TMA levels were found to be positively and significantly correlated with Klebsiella pneumoniae, suggesting that this bacterium is closely linked to the production of TMA in the elderly gut. This may have implications for the development and progression of CVD in the elderly population.

Transcriptomic Analysis of Wuzi Yanzongwan on Testicular Spermatogenic Function in Semi-castrated Male Mice / 中国实验方剂学杂志

ObjectiveTo screen out the transcriptomes related to the intervention of Wuzi Yanzongwan on the spermatogenic function of semi-castrated male mice， and to explore its potential mechanism in the intervention of the progress of low spermatogenic function. MethodBalb/c mice were randomly divided into sham-operated group， model group， testosterone propionate group（0.2 mg·kg-1·d-1， intramuscular injection） and Wuzi Yanzongwan group（1.56 g·kg-1·d-1， intragastric administration） according to body weight， with 12 mice in each group. The right testicle and epididymis were extracted from the model group and the drug administration group to construct the semi-castrated model of low spermatogenic function， while the fur and the right scrotum of the sham-operated group were only cut and immediately sterilized and sutured. At the end of the intervention， hematoxylin-eosin（HE） staining was used to observe the histopathology of testis， enzyme-linked immunosorbent assay（ELISA） was used to detect the levels of serum testosterone（T）， luteinizing hormone（LH） and follicle stimulating hormone（FSH）. The sperm count and motility of epididymis were measured by automatic sperm detector of small animal. Transcriptomic microarray technology was used to detect the mRNA expression level of testicular tissue in each group， the transcriptome of genes related to the regulation of Wuzi Yanzongwan was screened， and three mRNAs were selected for Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） to verify the transcriptome data. Through the annotation analysis of Gene Ontology（GO） and the signaling pathway analysis of Kyoto Encyclopedia of Genes and Genomes（KEGG）， the related functions of drugs regulating transcriptome were analyzed. ResultCompared with the sham-operated group， the testicular tissue of mice in the model group showed spermatogenic injury， contraction and vacuolization of the seminiferous tubules， reduction of spermatogenic cells at all levels， widening of the interstitial space， obstruction of spermatogonial cell development and other morphological abnormalities， and serum T significantly decreased， LH significantly increased（P<0.01）， and FSH elevated but no statistically significant difference， the count and vitality of epididymal sperm significantly decreased（P<0.01）. There were 882 differentially expressed mRNAs in the testicular tissues， of which 565 were up-regulated and 317 were down-regulated. Cluster analysis showed that these differentially expressed mRNA could effectively distinguish between the sham-operated group and the model group. Compared with the model group， the damage to testicular tissue in the Wuzi Yanzongwan group was reduced， the structure of the seminiferous tubules was intact， vacuolization was reduced， and the number of spermatogenic cells at all levels was significantly increased and arranged tightly. The serum T significantly increased， LH significantly decreased（P<0.01）， and FSH decreased but the difference was not statistically significant. The count and vitality of sperm in the epididymis were significantly increased（P<0.01）. Moreover， Wuzi Yanzongwan could regulate 159 mRNA levels in the testes of semi-castrated mice， of which 32 were up-regulated and 127 were down-regulated， and the data of the transcriptome assay was verified to be reliable by Real-time PCR. GO and KEGG analysis showed that the transcriptome functions regulated by Wuzi Yanzongwan were involved in the whole cell cycle process of sperm development such as sex hormone production of interstitial cells in testis， renewal， differentiation， metabolism， apoptosis and signal transduction of spermatogenic cells， and were closely related to the biological behaviors of signaling pathways such as spermatogenic stem cell function， endoplasmic reticulum protein processing and metabolic program. ConclusionWuzi Yanzongwan can effectively improve the low spermatogenic function of semi-castrated male mice， and its mechanism may be related to the regulation of testicular transcriptional regulatory network， the synthesis of sex hormones in testicular interstitial cells， the function of spermatogenic stem cells， the whole cell cycle process of spermatogenesis， as well as the expression of endoplasmic reticulum protein processing and metabolic program related genes transcription.

Expression level of epithelial cell adhesion molecule (EpCAM) of circulating tumor cells (CTCs) of patients with NSCLC as an early indicator to monitor the effects of postoperative adjuvant chemotherapy.

BACKGROUND: An early indicator for monitoring the effect of adjuvant treatment after lung cancer surgery is urgently needed. The study was to explore the effects of epithelial cell adhesion molecule (EpCAM) of circulating tumor cells (CTCs) in NSCLC patients with postoperative adjuvant chemotherapy. METHODS: Two drugs (platinum-containing chemotherapeutics + platinum-free chemotherapeutics) first-line chemotherapy regimen were given after surgery. MRNA of EpCAM was detected. Chest computed tomography, head computed tomography and abdominal B-ultrasound were reviewed before the first and third chemotherapy. RESULTS: EpCAM in CTCs from peripheral blood between the recurrent group and the non-recurrent group at 1 day before surgery, first, second and third adjuvant chemotherapy were no significant differences (P>0.05). Only one day before the fourth adjuvant chemotherapy treatment, it showed significant difference between the recurrent group and the non-recurrent group (P=0.008). There was a significant difference between the time of imaging diagnosis of recurrence or metastasis and the time of monitoring the expression level of EpCAM in CTCs from peripheral blood (P<0.0001). CONCLUSIONS: EpCAM in CTCs from peripheral blood during postoperative adjuvant chemotherapy was related to recurrence or metastasis of NSCLC patients.

[Expression and significance of microRNA-125b in tongue squamous cell carcinoma].

OBJECTIVE: The expression of microRNA-125b in tongue squamous cell carcinoma (TSCC) was detected and analyzed for its relationship with the clinicopathological features of TSCC. METHODS: Real time fluorescence-quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of microRNA-125b in 35 TSCC tissues and adjacent normal tissues from 35 TSCC cases. The relationship between the expression of microRNA-125b in TSCC tissues and the clinicopathological features of patients with TSCC was analyzed. In situ hybridization (ISH) was used to detect the expression level of microRNA-125b gene in the TSCC tissues and adjacent normal tissues. RESULTS: RT-qPCR results showed that the relative expression levels of microRNA-125b in the TSCC issues was 2.32±0.69, and that of normal tissues was 0.87±0.32. The statistical results showed that the expression level of microRNA-125b was significantly higher in the TSCC tissues than in the normal tissues (P<0.001). The expression level of microRNA-125b in the TSCC tissues was not significantly correlated with age, gender, pathological grade, and lymph node metastasis but was positively correlated with TNM stage. Patients with high TNM stage had high microRNA-125b expression levels (P<0.05). The ISH results showed that the expression levels of microRNA-125b in the TSCC tissues were 0.010±0.003, and that of normal tissues was 0.004±0.001. The expression levels of microRNA-125b in the 35 TSCC tissues were significantly higher than those in the normal tissues (P<0.05). CONCLUSIONS: MicroRNA-125b is highly expressed in TSCC and associated with TNM stage, suggesting that high microRNA-125b expression may be involved in the development of TSCC.

Development and Comparison of Determination of Staphylococcus aureus in Different Processed Products of Angelicae Sinensis Radix Based on Real-time PCR Technique / 中国实验方剂学杂志

Objective:To quantitatively analyze the changes of Staphylococcus aureus in different processed products of Angelicae Sinensis Radix. Method:The real-time fluorescence quantitative polymerase chain reaction method （Real-time PCR） was established to quantitatively analyze S. aureus in Angelicae Sinensis Radix decoction pieces which bought from different producing areas， different enterprises and different storage time. The fluorescence quantitative reaction system was SYBR Premix Ex Taq Ⅱ of 10 μL， each of forward primer and reverse primer （10 μmol·L-1） of 0.8 μL， template/genome DNA of 1 μL， double distilled water of 7.4 μL. The reaction conditions of the fluorescence quantitative amplification curve were pre-denaturing for 30 s at 94 ℃， denaturing for 10 s at 94 ℃， annealing for 12 s at 60 ℃， extensing for 30 s at 72 ℃， cycling 45 times， single-point detection signal at 72 ℃. The melting curve was made from 72 ℃， and the step temperature of 0.5 ℃ was kept for 15 s to collect fluorescence. According to the results of Real-time PCR， representative samples were selected from Angelicae Sinensis Radix decoction pieces for comparison between plate counting method and Real-time PCR. Result:The content of S. aureus in different processed products was sorted by rank of raw Angelicae Sinensis Radix>soil-fried Angelicae Sinensis Radix>wine-processed Angelicae Sinensis Radix. The content of S. aureus was the lowest in the samples from Weiyuan area of Gansu province by comparing with other producing areas. Compared with the retail enterprises， the content of S. aureus in raw products and wine-processed products from production and sale enterprises was lower. Different storage time had certain effect on the content of S. aureus in raw products and wine-processed products， and the content of S. aureus increased with the increase of storage time. The detection results of plate counting method were 3-4 orders of magnitude lower than that of Real-time PCR. Conclusion:The established Real-time PCR is superior to plate counting method in specificity， sensitivity， reliability and reporting period， which can provide an effective method for rapid and accurate quantitative detection of S. aureus in different processed products of Angelicae Sinensis Radix.

Expression and significance of microRNA-125b in tongue squamous cell carcinoma / 华西口腔医学杂志

OBJECTIVE@#The expression of microRNA-125b in tongue squamous cell carcinoma (TSCC) was detected and analyzed for its relationship with the clinicopathological features of TSCC.@*METHODS@#Real time fluorescence-quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of microRNA-125b in 35 TSCC tissues and adjacent normal tissues from 35 TSCC cases. The relationship between the expression of microRNA-125b in TSCC tissues and the clinicopathological features of patients with TSCC was analyzed. In situ hybridization (ISH) was used to detect the expression level of microRNA-125b gene in the TSCC tissues and adjacent normal tissues.@*RESULTS@#RT-qPCR results showed that the relative expression levels of microRNA-125b in the TSCC issues was 2.32±0.69, and that of normal tissues was 0.87±0.32. The statistical results showed that the expression level of microRNA-125b was significantly higher in the TSCC tissues than in the normal tissues (P<0.001). The expression level of microRNA-125b in the TSCC tissues was not significantly correlated with age, gender, pathological grade, and lymph node metastasis but was positively correlated with TNM stage. Patients with high TNM stage had high microRNA-125b expression levels (P<0.05). The ISH results showed that the expression levels of microRNA-125b in the TSCC tissues were 0.010±0.003, and that of normal tissues was 0.004±0.001. The expression levels of microRNA-125b in the 35 TSCC tissues were significantly higher than those in the normal tissues (P<0.05).@*CONCLUSIONS@#MicroRNA-125b is highly expressed in TSCC and associated with TNM stage, suggesting that high microRNA-125b expression may be involved in the development of TSCC.

Expression of CCL18 gene in ovarian cancer and its impact on the biologic function of ovarian cancer cells.

The aim of the present study was to evaluate the expression of the chemokine ligand 18 (CCL18) gene in ovarian cancer and to investigate the effects of its overexpression or suppression on growth, invasion, and metastasis in an ovarian carcinoma cell line (SKOV3) in vitro. CCL18 mRNA expression in epithelial ovarian carcinoma (EOC), benign ovarian tumor and normal ovarian tissues was measured by fluorescence quantitative polymerase chain reaction. A CCL18 restructuring plasmid was constructed, and SKOV3 cells were transfected with the plasmid DNA in vitro. A restructuring interference vector was also transfected into CCL18-positive SKOV3 cells. The growth curves, cell cycle distribution, and invasive, migrative and adhesive capacities of SKOV3 cells following overexpression and suppression of CCL18 were evaluated by MTT assay, flow cytometry, Transwell assay, migration assay, and the fibronectin adhesion method, respectively. The positive expression rate of CCL18 in EOC was significantly higher than in benign ovarian tumor (P = 0.002) and normal ovarian tissues (P = 0.003). However, there was no statistical significance in the expression of CCL18 with regard to clinical pathology (including histological classification, pathological grade and surgical pathological stage), and the median survival times of CCL18-positive and CCL18-negative patients did not differ significantly. The invasive, migrative, and adhesive capacities of SKOV3-CCL18 cells were significantly higher than those of SKOV3 and SKOV3-vector cells (P < 0.05). However, there was no significant difference in cell proliferation between the SKOV3-CCL18 and negative control cells. The invasive, migrative, and adhesive capacities of the pSilencer4.1-CCL18-small interfering RNA127 group were significantly lower than those of non-transfected pSilencer4.1-negative and pSilencer4.1 groups (P < 0.05). In conclusion, the overexpression and silencing of CCL18 affected invasion, adhesion, and migration in EOC cells; thus CCL18 may have potential as a clinical marker for early diagnosis of malignant ovarian tumors, and as a target molecule in the treatment of ovarian cancer.

Preliminary Research on Dynamic Change of Microbial Population in Fermentation Process of Pinelliae Rhizoma Fermentata / 中国实验方剂学杂志

Objective:To detect the colony number of bacteria, yeasts and molds in fermentation process of Pinelliae Rhizoma Fermentata (PRF), microbial flora species, and quantitatively analyze the dynamic changes of four dominant microorganisms at different fermentation time points of PRF, so as to provide experimental basis for exploring the processing mechanism of PRF. Method:According to Pharmaceutical Standard Preparation of Traditional Chinese Medicine Prescription of Ministry of Health of the People's Republic of China (the 10th volume), PRF was processed. The samples at five different fermentation time points (0, 30, 60, 90, 120 h) of PRF were taken, the culturing, isolation and purification of bacteria, yeasts and molds were carried out with selective media, and the colonies were counted. Fluorescence quantitative polymerase chain reaction (PCR) technique was employed to conduct absolute quantification of Bacillus subtilis, Paecilomyces variotii, Byssochlamys spectabilis and Aspergillus niger. The recombinant plasmids of these 4 microorganisms were used as the standard substances, and the standard curves were prepared after dilution of multiple ratios, quantitative analysis was performed on these 4 microorganisms in five samples at different processing time points (0, 30, 60, 90, 120 h) of PRF. Result:During the fermentation process of PRF, the number of bacteria was low with smooth change, while molds and yeasts grew dramatically at the late stage of fermentation and reached 1×106 CFU·mL-1 at the end of fermentation. At 5 different fermentation time points, the copy numbers of Bacillus subtilis were 3.53×105, 7.56×104, 1.58×105, 1.90×106, 1.85×106 copies·g-1, the copy numbers of Paecilomyces variotii were 0, 0, 0, 3.45×107, 4.15×108 copies·g-1, the copy numbers of Byssochlamys spectabilis were 0, 0, 0, 1.04×108, 2.28×108 copies·g-1, the copy numbers of Aspergillus niger were 0, 0, 9.48×105, 1.47×106, 7.56×106 copies·g-1, respectively. Conclusion:The change trend of microflora in the fermentation process of PRF can be reflected by the dynamic change of four dominant microorganisms, and molds may play an important role in the processing of PRF. Fluorescence quantitative PCR technique has the advantages of rapidity, sensitivity, good repeatability and high specificity, it is suitable for exploring processing mechanism of PRF.

Expression of RIZ1 gene in myelodysplastic syndromes and its significance / 肿瘤研究与临床

Objective To explore RIZ1 mRNA expression level in different subtypes and risk classification groups of the patients with myelodysplastic syndromes (MDS) and to analyze the correlation between the clinical features and RIZ1 mRNA expression. Methods A total of 46 newly diagnosed as MDS patients and 10 healthy controls who were the donors of hematopoietic stem cell transplantation from Chuiyangliu Hospital Affiliated to Tsinghua University and the Second Hospital of Shanxi Medical University between January 2014 and December 2017 were collected. The real time fluorescence quantitative polymerase chain reaction (RT-PCR) was used to detect the expression level of RIZ1 mRNA in bone marrow cells from MDS patients and the healthy controls. Results Compared with the healthy control group, the relative expression level of RIZ1 mRNA in MDS patients was decreased [the median (P25, P75):1.003 (0.895, 1.812) vs. 0.557 (0.333, 0.815)], and the difference was statistically significant (Z= -2.991, P= 0.0003). According tothe World Health Organization (WHO) classification criteria, compared with the healthy control group, the relative expression of RIZ1 mRNA in refractory anemia/refractory anemia with ring sideroblasts/refractory cytopenia with multiple dysplasia (RA/RAS/RCMD), refractory anemia with excess blasts Ⅰ (RAEB-Ⅰ), RAEB-Ⅱand MDS transformed into acute myeloid leukemia (MDS/AML) groups had statistically significant differences (χ2= 19.500, P< 0.01). Further pairwise comparison showed that the relative expression level of RIZ1 mRNA in RAEB-Ⅰ, RAEB-Ⅱand MDS/AML groups was lower than that in the healthy control group, and the differences were statistically significant (all P < 0.05). According to the international prognostic scoring system (IPSS), compared with the healthy control group, the expression of RIZ1 mRNA in low-risk, inter-risk-1, inter-risk-2 and high-risk group had statistical differences (χ2= 19.214, P= 0.001). Further pairwise comparison showed that the relative expression of RIZ1 mRNA in inter-risk-1, inter-risk-2 and high risk group was lower than that in the healthy control group, and the differences were statistically significant (all P<0.05). And RIZ1 mRNA expression showed a decreasing trend with the increase of disease risk grade. RIZ1 mRNA expression level had no relationship with age, gender, peripheral blood white cell count, the hemoglobin, platelet count and karyotype (all P> 0.05). Conclusion RIZ1 mRNA expression level is decreased in MDS patients, and it is different in various subtypes and risk classification. RIZ1 may involve in the pathogenesis of MDS and play an important role in the progression of MDS.

Levels of miRNA－106a in non－small cell lung cancer tissues and its clinical significance / 中国基层医药

Objective To investigate the level and clinical significance of miRNA －106a in non－small cell lung cancer tissues.Methods Paraffin－embedded specimens of non －small cell lung cancer tissues and adjacent tissues from 80 patients with non －small cell lung cancer from January 2012 to December 2013 in Yiwu Central Hospital were selected in this study.The real－time fluorescence quantitative polymerase chain reaction (QRT－PCR) was used to determine the level of miRNA －106a in lung cancer tissues and adjacent tissues.Results The level of miRNA－106a in non－small cell lung cancer tissues (2.42 ±0.23) was higher than that in adjacent tissues (1.00 ± 0.06) (t＝53.433,P＝0.000).The miRNA －106a levels in lung cancer tissues of stage Ⅲ －Ⅳ,lymph node metastasis and recurrence time ＜6 months were high than those of the stage Ⅰ－Ⅱ,no lymph node metastasis and recurrence time≥6 months(t＝7.641,11.115,2.183,P＝0.000,0.000,0.032).The level of miRNA－106a was not associated with age ,gender,pathological type,degree of differentiation and vascular invasion (all P＞0.05).The level of miRNA－106a in non－small cell lung cancer tissues was cut by 2.12.The area under the ROC curve for the diagnosis of non －small cell lung cancer was 0.823(95%CI＝0.820－0.825,P＝0.000),and the sensitivity was 54.37%,the specificity was 89.21%.The cumulative survival rate and progression －free survival rate of patients with low－expression of miRNA －106a in non －small cell lung cancer were higher than those with high expression of miRNA－106a (P＝0.027,0.012).Conclusion The miRNA －106a level is elevated in non －small cell lung cancer tissues.The miRNA－106a may be involved in the development of non －small cell lung cancer and is of great value in the diagnosis and prognosis evaluation of non －small cell lung cancer.

Levels of miRNA-106a in non-small cell lung cancer tissues and its clinical significance / 中国基层医药

Objective@#To investigate the level and clinical significance of miRNA-106a in non-small cell lung cancer tissues.@*Methods@#Paraffin-embedded specimens of non-small cell lung cancer tissues and adjacent tissues from 80 patients with non-small cell lung cancer from January 2012 to December 2013 in Yiwu Central Hospital were selected in this study.The real-time fluorescence quantitative polymerase chain reaction (QRT-PCR) was used to determine the level of miRNA-106a in lung cancer tissues and adjacent tissues.@*Results@#The level of miRNA-106a in non-small cell lung cancer tissues (2.42±0.23) was higher than that in adjacent tissues (1.00±0.06) (t=53.433, P=0.000). The miRNA-106a levels in lung cancer tissues of stage Ⅲ-Ⅳ, lymph node metastasis and recurrence time<6 months were high than those of the stage Ⅰ-Ⅱ, no lymph node metastasis and recurrence time ≥ 6 months(t=7.641, 11.115, 2.183, P=0.000, 0.000, 0.032). The level of miRNA-106a was not associated with age, gender, pathological type, degree of differentiation and vascular invasion (all P>0.05). The level of miRNA-106a in non-small cell lung cancer tissues was cut by 2.12.The area under the ROC curve for the diagnosis of non-small cell lung cancer was 0.823(95% CI=0.820-0.825, P=0.000), and the sensitivity was 54.37%, the specificity was 89.21%.The cumulative survival rate and progression-free survival rate of patients with low-expression of miRNA-106a in non-small cell lung cancer were higher than those with high expression of miRNA-106a (P=0.027, 0.012).@*Conclusion@#The miRNA-106a level is elevated in non-small cell lung cancer tissues.The miRNA-106a may be involved in the development of non-small cell lung cancer and is of great value in the diagnosis and prognosis evaluation of non-small cell lung cancer.

Micro-droplet Digital Polymerase Chain Reaction and Real-Time Quantitative Polymerase Chain Reaction Technologies Provide Highly Sensitive and Accurate Detection of Zika Virus.

The establishment of highly sensitive diagnostic methods is critical in the early diagnosis and control of Zika virus (ZIKV) and in preventing serious neurological complications of ZIKV infection. In this study, we established micro-droplet digital polymerase chain reaction (ddPCR) and real-time quantitative PCR (RT-qPCR) protocols for the detection of ZIKV based on the amplification of the NS5 gene. For the ZIKV standard plasmid, the RT-qPCR results showed that the cycle threshold (Ct) value was linear from 101 to 108 copy/µL, with a standard curve R2 of 0.999 and amplification efficiency of 92.203%; however, a concentration as low as 1 copy/µL could not be detected. In comparison with RT-qPCR, the ddPCR method resulted in a linear range of 101-104 copy/µL and was able to detect concentrations as low as 1 copy/µL. Thus, for detecting ZIKV from clinical samples, RT-qPCR is a better choice for high-concentration samples (above 101 copy/µL), while ddPCR has excellent accuracy and sensitivity for low-concentration samples. These results indicate that the ddPCR method should be of considerable use in the early diagnosis, laboratory study, and monitoring of ZIKV.

Establishment and evaluation of different serotypes group B streptococcus method by TaqMan fluorescent probe technique / 国际检验医学杂志

Objective To establish a molecular method for the identification of different serotypes of group B streptococcus(GBS)based on TaqMan fluorescence probe technology,and to lay the foundation for the sub-sequent study of multiple fluorescent probe technology to detect different serotypes of GBS.Methods Primers and probes were designed according to the different serotypes of capsular polysaccharide(CPS).CPS se-quences were amplified by real-time fluorescence quantitative polymerase chain reaction.GBS classification methods of different serotypes were established.The results were compared with latex agglutination test and the method was evaluated from the aspects of sensitivity,specificity and detection of clinical isolates.Results The logarithmic concentration of DNA in the same serotype GBS was linearly correlated with the value of Ct. The detection limit of this method is 1 pg/μL,a probe could only detect the corresponding serotype GBS.The results of TaqMan fluorescence probe test of 10 strains were consistent with the results of latex agglutination test.Conclusion TaqMan fluorescence probe technique is a simple,rapid,highly sensitive and specific method for the detection of different GBS serotypes,and it is better than latex agglutination test for the classification of clinical isolates.

Study on CMV-DNA detection in blood and urine / 国际检验医学杂志

Objective To study the detection of cytomegalovirus (CMV)-DNA in different kinds of blood and urine sample .Methods The CMV-DNA loads in 3 different kinds of blood sample from 52 patients and 3 different kinds of urine sample from 85 patients were detected by real-time fluorescence quantitative polymer-ase chain reaction(FQ-PCR) .The differences in CMV-DNA detection rate and virus loads were compared a-mong different kinds of blood and urine samples .Results The CMV-DNA detection rates in serum ,whole blood ,plasma and peripheral blood mononuclear cell (PBMC) from 52 patients were 48 .08％ ,71 .15％ ,57 .69％ and 69 .23％ respectively .The CMV-DNA detection rates of whole blood and PBMC were higher ,the difference was statistically significant (P<0 .05) .The quantitative results of PBMC was higher .The CMV-DNA detec-tion rates of mixed urine ,urine supernatant and urine sediment from 85 patients were 72 .94％ ,62 .35％ and 84 .71％ respectively ,the CMV-DNA detection rate of urine sediment was higher ,while the quantitative re-sults of mixed urine was higher .Conclusion The CMV-DNA detection results of blood and urine have large difference ,therefore using the same kind of sample for conducting detection has an important clinical signifi-cance in the clinical diagnosis ,treatment and monitoring of CM V infection .

Anti-fatigue effect of arginyl fructosyl glucose from red ginseng in mice / 吉林大学学报(医学版)

Objective:To investigate the anti-fatigue phenomenon induced by forced swimming in the mice,and to explore the anti-fatigue effect of argininyl fructoyl glucose (AFG) from red ginseng in the mice and its mechanism.Methods:The AFG was extracted from red ginseng.The ICR mice were divided into blank control group,low dose of AFG group (100 mg · kg-1),middle dose of AFG group (200 mg · kg-1) and high dose of AFG group (400 mg · kg-1) (n=20).The mice mere given a forced swimming test after continuous gavage for 28 d.The weights,organ indexes,time of forced swimming,contents of lactic acid (LD),blood urea nitrogen (BUN),hepatic glycogen (Gly) and expressing levels of PGC-1α in gastrocnemius of the mice in various groups were detected.Results:Compared with blank control group,the weights and organ indexes of the mice in low,middle and high doses of AFG groups had no significant differences (P＞ 0.05).Compared with blank control group,the time of forced swimming,contents of Gly and expressing levels of PGC-1α of the mice in low,middle and high doses of AFG groups were significantly increased (P＜0.05 or P＜0.01) in a dose-dependent manner.Compared with blank control group,The contents of LD and BUN in serum of the mice in low,middle and high doses of AFG groups were significantly decreased (P＜0.01).Conclusion:AFG has anti-fatigue effect in mice,and its mechanism may be related to energy metabolism.

Construction of standard plasmids for detecting Salmonella in meat products with real-time fluorescence quantitative polymerase chain reaction / 上海预防医学

Objective To construct standard plasmids for detecting Salmonella in meat products with real-time fluorescence quantitative polymerase chain reaction(PCR).Methods Primers directed at Salmonella invA gene were designed.Specific fragments were amplified and cloned into plasmid vectors to construct recombinant plasmids.The constructed recombinant plasmids were used as standards for implementing the real-time fluorescence quantitative PCR after optimization, establishing standard curves and examining the sensitivity and stability of these standards.Results The plasmid standard containing the invA gene in Salmonella was successfully constructed.The cycle threshold value(Ct value) of the standard curve established by means of the plasmid standard and the number of template copies exhibited good linear relationship(r2=0.9979).The minimum that could be detected by means of this method was 10 copies/reaction.The standard plasmid was proved to have good stability.This standard plasmid was used to detect Salmonella in meat products.After two hours enrichment, sample detection could be completed within seven hours.Conclusion The constructed plasmid standard can be used for detecting Salmonella in meat products by means of the fluorescence quantitative PCR, which can provide reliable reference bases for examining quality of relevant experiments.

Anti-fatigue effect of arginyl fructosyl glucose from red ginseng in mice / 吉林大学学报(医学版)

Objective:To investigate the anti-fatigue phenomenon induced by forced swimming in the mice,and to explore the anti-fatigue effect of argininyl fructoyl glucose (AFG) from red ginseng in the mice and its mechanism.Methods:The AFG was extracted from red ginseng.The ICR mice were divided into blank control group,low dose of AFG group (100 mg · kg-1),middle dose of AFG group (200 mg · kg-1) and high dose of AFG group (400 mg · kg-1) (n=20).The mice mere given a forced swimming test after continuous gavage for 28 d.The weights,organ indexes,time of forced swimming,contents of lactic acid (LD),blood urea nitrogen (BUN),hepatic glycogen (Gly) and expressing levels of PGC-1α in gastrocnemius of the mice in various groups were detected.Results:Compared with blank control group,the weights and organ indexes of the mice in low,middle and high doses of AFG groups had no significant differences (P＞ 0.05).Compared with blank control group,the time of forced swimming,contents of Gly and expressing levels of PGC-1α of the mice in low,middle and high doses of AFG groups were significantly increased (P＜0.05 or P＜0.01) in a dose-dependent manner.Compared with blank control group,The contents of LD and BUN in serum of the mice in low,middle and high doses of AFG groups were significantly decreased (P＜0.01).Conclusion:AFG has anti-fatigue effect in mice,and its mechanism may be related to energy metabolism.

The value of quantitative PCR detection in early skin tissue fluid of syphilis / 国际检验医学杂志

Objective To study the value of fluorescence quantitative polymerase chain reaction (FQ-PCR) detection in early skin tissue fluid of syphilis.Methods A total of 40 patients of suspected syphilis who received therapy from September 2014 to September 2016 in our hospital were selected in this study.Five mL venous blood samples were collected in all the patients,and detected by toluidine red unheated serum test (TRUST) and treponema pallidum antibody enzyme-linked immunosorbent assay (ELISA),and skin tissue fluid were collected and performed FQ-PCR detection,all patients were treated with benzylpenicillin for 3 and 6 months,then detected again,the conversion rates were record.Results There was no significant difference in the detection rate of ELISA and FQ-PCR[97.50%(39/40) vs.95.00%(38/40),P>0.05].The total detection rate of ELISA and FQ-PCR were significantly higher than that of TRUST[97.50%(39/40) vs.67.50%(27/40),95.00%(38/40) vs.67.50%(27/40),P0.05).In the FQ-PCR detection results,the average value of TP-DNA was significantly decreased after treatment,there were significant differences in the phase Ⅰ,phase Ⅱ compared with before treatment(P<0.05).Conclusion FQ-PCR could be used to measure treponema pallidum (TP-DNA) effectively in early stage,it′s conducive to the diagnosis of syphilis,the clinical application value is high.

Clinical application of real-time fluorescence quantitative PCR for the detection of fecal tumor M2-pyruvate kinase in colorectal cancer patients / 国际检验医学杂志

Objective To investigate the application valve of real-time fluorescence quantitative polymerase chain reaction(RT-PCR) for the detection of tumor M2-pyruvate kinase(tM2-PK) DNA in patients with colorectal cancer(CRC).Methods Fragment of tM2-PK DNA(162 bp) was amplified and inserted into PGM-T vector to construct recombinant plasmid,which was used to develop RT-PCR method.Sensitivity,specificity and repeatability of RT-PCR for the detection of tM2-PK were analyzed.From Jan.2014 to Jun.2016,200 CRC patients and 100 healthy subjects were enrolled and detected for fecal and serum tM2-PK DNA by using RT-PCR,and the detected results were compared with those detected by using enzyme linked immunosorbent assay(ELISA).Results Recombinant plasmid was successfully constructed,which was certified by sequencing.The sensitivity of RT-PCR for the detection of tM2-PK DNA was 10 copy/mL,with high specificity and 0.3%-2.9% of coefficient of variation.In patients,the positive rate of fecal tM2-PK DNA,detected by RT-PCR,was 92.50%,and that of ELISA to detect tM2-PK was 80.00%.Fecal and serum levels of tM2-PK were correlated with the pathologic stages of tumour.Conclusion Self-established RT-PCR could be specificity and sensitivity for the detection of fecal tM2-PK,which could be used for the early diagnosis of CRC.

The expression of protection of telomeres 1 in acute myeloid leukemia and its significance / 白血病·淋巴瘤

Objective To analyze the relationship between the expression of protection of telomeres 1 (POT1) and the pathogenesis of acute myeloid leukemia (AML). Methods 62 patients with de novo AML (case group) and 10 patients with iron deficiency anemia (control group) were enrolled in this study. The quantitative real-time polymerase chain reaction (PCR) and Western blot were used to detect the expression of POT1 in AML patients. Results There were 62 de novo AML patients, including 2 cases M1, 14 cases M2, 12 cases M3, 14 cases M4, 17 cases M5, 2 cases M6 and 1 case AML without classification. According to the risk stratification, high risk group (24 cases), medium risk group (22 cases) and low risk group (16 cases) were divided. Compared with that in the controls, POT1 expression levels in patients with AML were significantly decreased both in mRNA and protein level (P 0.05). Conclusions POT1 may be involved in the pathogenesis of AML. POT1 protein expresses in both cytoplasm and nucleus, and the regulatory mechanism may be related to the telomere length.