Objective To develop SYBR Green I real-time PCR assay for detection and identification of Hepatitis B virus. Methods Based on the sequences of Hepatitis B virus gp1 gene,primers were designed.The reaction assay and thermal cyc-ling profile were optimized.The positive standard was from recombinant clone.Both the developed assay and Zhejiang kuake biotechnology company’s assay were applied in 100 patients serum.Results The detection limit was between 5×102 copies/ml to 5×108 copies/ml with a good liner correlation and no cross reaction.The whole process just needed 2.5 h.Comparing with the company products,the sensitivity and specificity of the developed assay were 100% and 92.5% respectively.Con-clusion The established assay is rapid,simple,high sensitivity and specificity.It is not only valuable for the identification of Hepatitis B virus patients,but also provide accurate quantitative analysis for HBV patients.

A HPLC-QTOF MS method was established for analysis of components in Ophiocordyceps sinensis . The HPLC analysis was performed on an Agilent Zorbax SB Aq (150 mmí4.6 mm, 5 μm) with gr adient elution (5 mmol·L-1 ammonium acetate aqueous solution-acetonitrile), flow rate was 0.8 mL·min-1 and detection wave-length was 260 nm. The developed method was successfully applied in analysis of three different samples in-cluding O. sinensis, Hirsutella sinensis ( anamorph of O.sinensis) and Cordyceps militaris. Nine compounds were i-dentified in both O.sinensis and H.sinensis, which eight compounds were identified in C.militaris.

Objective To explore the clinical effect of Jiangru Formula(Herbal formula for soothing the liver,clearing heat,activating blood and removing stasis)combined with TCM surgical treatment for plasmocyte mastitis.Methods Fifty five patients with plasmocyte mastitis were treated with Jiangru Formula orally and TCM surgical treatment to see the effect of this method.Results The total effective rate was 100%.The average treatment period was(28?13.61)days.The recurrence rate was 1.82%.Conclusion The combination of Jiangru Formula with TCM surgical treatment for plasmocyte mastitis is effective in shortening the treatment period,reducing the recurrence rate and keeping the shape of the breast.

Objective To develop TaqMan real-time PCR assay for detection and identification of streptococcus pneumonia iso-lated from children CAP.Methods Based on the sequences of lytA gene,primers and probe were designed and the assay was optimized.Then 1 504 sputum samples were detected by culture and the developed assay with double-blind testing.Results The lower limit of detection in the developed real-time PCR assay was 18.75 cfu/PCR,and had no cross reaction.141 strep-tococcus pneumonia strains were detected from 1 504 samples and 140 were isolated by culture.The whole process just nee-ded 2.5 h.Conclusion The established assay is rapid,simple,high sensitivity and specificity.It is not only valuable for the i-dentification of streptococcus pneumonia,but also provide evidence for antibiotic therapy.

Objective To explore the effect of anti-retroviral therapy on interleukin(IL)-7/IL-7R in human immunodeficiency virus(HIV) infected patients in China.Methods Cases were divided into 2 groups:HIV-infected group (35 cases),and control group (30 cases).IL-7 in serum,IL-7R(CD127) expression in CD4 +T cells,and CD4 +T cells count were detected and compared between two groups before and after treatment for 1 year.Results IL-7 level in the serum of HIV infected group before treatment [(8.98 ±3.77) pg/ml] was significantly higher than that in control group [(3.84 ±0.86) pg/ml] (P ＜0.05).The counts of CD4+T cells [(202.65 ± 121.54)/μl],CD4 + CD127 + T cells [(60.25 ± 11.75) ％],and CD8 + CD127 + T cells [(46.27 ± 12.10)％] in HIV-infected group were significantly lower than those in control group [(766.99 ± 103.21)/L,(76.89 ± 20.01) ％,(81.27 ± 12.35)％] (P ＜0.05).After anti-retroviral therapy (ART),IL-7 level in the serum of HIV-infected group[(5.55 ± 1.35) pg/ml]was decreased,and CD4+T cells [(450.58 ± 15)/μl],CD4 + CD127 +T cells [(69.82 ± 15.24)％],and [CD8 + CD127 + T(59.23± 14.73) ％] cells was increased in HIV-infected group,with a significant difference between two groups (P ＜0.05).Conclusions ART could improve the IL-7 level in the serum and IL-7R(CD127)expression in CD4 +T cells of HIV-infected patients.However,they still cannot become normal level.

Objective To explore the efficacy and side effects of highly active antiretroviral therapy (HAART) in human immunodeficiency virus (HIV)-infected patients with different CD4+ cell counts.Methods The clinical data of HIV-infected patients who accepted TDF (tenofovir disoproxil fumarate) + 3TC (lamivudine) + EFV (efavirenz) treatment were retrospectively collected in Jiangmen region.All patients were divided into group A(350/μl ≤ CD4 + ＜ 500/μ1),group B (200/μl ≤ CD4 + ＜ 350/μl) and group C(CD4+ ＜ 200/μl)according to their CD4+ cell counts.The efficacy and side effects in different groups were compared.Results A total of 132 clinical cases was collected,including 32 cases in group A,42 cases in group B,and 58 cases in group C.No statistically difference was found among three groups in terms of gender,age,or route of transmission.CD4 + cell counts after treatment was significantly higher than that before treatment in each group (P ＜ 0.05).The increase of CD4 + cell counts in groups A,B,and C was 75.6 ± 52.1,80.5 ± 58.7,and 97.5 ± 78.7 after 6-month HAART,respectively ; and 71.4 ± 58.9,110.8 ± 71.6,and 113.7 ± 88.3 after 12-month HAART,respectively.Statistical analysis showed no significant difference among three groups (P ＞ 0.05).The incidence of side effects in groups A,B,and C was 4/32,14/42,and 32/58 in 3-month HAART,respectively; and 1/32,8/42,and 22/58 in 3 ～ 12 month HAART,respectively.Statistical analysis showed significant difference among three groups (group C ＞ group B ＞ group A,P ＜ 0.05).Conclusions It was effective to begin the anti-retroviral treatment in all stages.The incidence of side effects may be less if anti-retroviral treatment began in early period.

Aim To make a research of rats with focal cerebral ischemia/reperfusion injury on pathological changes in brain and the changes of AQP4 and related proteins, in order to explore the relationship between AQP4 and brain edema. Methods Adult male SD rats, weighting 250~300 g, were randomly divided in-to Sham group and cerebral ischemia/reperfusion ( I/R) injury model group. The I/R model group was di-vided into the I/R-6 h, 12 h, 24 h, 48 h-four time point groups. The animal model of the right MCA is-chemia/reperfusion was established by suture method in mature SD rats. The nerve symptom score was con-ducted in the corresponding time points. Then, the permeability of brain tissue was detected by EB stai-ning;TTC staining was conducted to observe the cere-bral infarction volume;the dry wet weight method was used to detect the changes of brain water content; im-munohistochemical( IHC) , WB and RT-PCR were ap-plied to detect the expression of AQP4 , and the related factors at different time points of the model rats after is-chemia-reperfusion around infarcts. Results Com-pared with the Sham group, then ever function score of the rats in I/R model groups were much higher. With the increase of the reperfusion time, the cerebral in-farction volume, brain tissue permeability and the brain water content were also increased. IHC results showed that AQP4 expression gradually rose with widen distribution. WB and RT-PCR results verified the in-creasing level of AQP4 expression. The detection of the related proteins expression showed apparent changes. The expression of MMP-9 was increased, while the Oc-cludin and JAM-1 expression showed a decreasing trend. The I/R-48 h model group showed the most ob-vious differences in the expression of the related pro-teins and mRNA ( P <0. 01 vs Sham, respectively ) . Conclusion Accompanied with the aggravating cere-bral injury after cerebral ischemia/reperfusion, the ex-pression of AQP4 and MMP-9 level were activated, while the degradation of TJPs, Occludin and JAM-1, was increased. These factors are combined to make the formation of brain edema. This study makes a further research on the formation mechanism of the early stage for cerebral edema on I/R model and offers a potential for intervention in the filed of looking for a reliable drug therapy on cerebral edema.

Objective:To evaluate the clinical value of abdominal color Doppler ultrasonography in the differential diagnosis of colonic polyps in children.Methods:From January 2017 to January 2020, 300 children with suspected colonic polyps were studied.All patients were examined by colonoscopy and abdominal color Doppler ultrasound respectively.The results of colonoscopy were taken as the gold standard to evaluate the diagnostic efficacy of abdominal color Doppler ultrasound in the differential diagnosis of colonic polyps in children.The diagnostic accuracy of abdominal color Doppler ultrasound in diagnosis of colonic polyps in children with different location, type and focus size was analyzed.Results:The diagnostic sensitivity and specificity of abdominal color Doppler ultrasound for colonic polyps in children were 87.60%(113/129) and 97.08%(166/171), respectively.The overall diagnostic accuracy was 93.00% (279/300), the positive predictive value was 95.76%(113/118), and the negative predictive value was 91.21% (166/182). The diagnostic accuracy of abdominal color Doppler ultrasound were 85.71%(12/14), 83.33%(10/11), 87.50%(28/32), 86.79%(46/53) and 89.47%(17/19) for children's polyps in ascending colon, transverse colon, descending colon, sigmoid colon and rectum, and were 86.17%(81/94), 92.31%(24/26) and 88.89%(8/9) for juvenile polyps, black spot polyp syndrome and adenomatous polyps, respectively.There were no statistically significant differences between the two groups (χ 2=0.250, 0.721; P=0.993, 0.697). The diagnostic accuracy of abdominal color Doppler ultrasound in children with diameter ≤1 cm (38.09%) was significantly lower than that in children with 1 cm

PURPOSE: Uridine-cytidine kinase (UCK) 2 is a rate-limiting enzyme involved in the salvage pathway of pyrimidine-nucleotide biosynthesis. Recent studies have shown that UCK2 is overexpressed in many types of cancer and may play a crucial role in activating antitumor prodrugs in human cancer cells. In the current study, we evaluated the potential prognostic value of UCK2 in breast cancer. METHODS: We searched public databases to explore associations between UCK2 gene expression and clinical parameters in patients with breast cancer. Gene set enrichment analysis (GSEA) was performed to identify biological pathways associated with UCK2 gene expression levels. Survival analyses were performed using 10 independent large-scale breast cancer microarray datasets. RESULTS: We found that UCK2 mRNA expression was elevated in breast cancer tissue compared with adjacent nontumorous tissue or breast tissue from healthy controls. High UCK2 levels were correlated with estrogen receptor negativity (p<0.001), advanced tumor grade (p<0.001), and poor tumor differentiation (p<0.001). GSEA revealed that UCK2-high breast cancers were enriched for gene sets associated with metastasis, progenitor-like phenotypes, and poor prognosis. Multivariable Cox proportional hazards regression analyses of microarray datasets verified that high UCK2 gene expression was associated with poor overall survival in a dose-response manner. The prognostic power of UCK2 was superior to that of TNM staging and comparable to that of multiple gene signatures. CONCLUSION: These findings suggest that UCK2 may be a promising prognostic biomarker for patients with breast cancer.
Biomarkers ; Breast Neoplasms* ; Breast* ; Dataset ; Estrogens ; Gene Expression ; Humans ; Neoplasm Metastasis ; Neoplasm Staging ; Phenotype ; Prodrugs ; Prognosis* ; RNA, Messenger ; Uridine Kinase*

This study was aimed to investigate the estrogenic effects ofCoicis Semen in order to preliminarily discuss the mechanism. Mouse uterine weight test and MCF-7 cell proliferation assay were used to evaluate the estrogenic effects ofC. Semen. Reporter gene assays were adopted to explore the action mechanism ofC. Semen. In reporter gene assay, HEK293 cells were co-transfected with pERE-TAL-luc, pβgal-Control, pCXN2-hERα, or pCXN2-hERβ. And the expression of reporter gene luc was controlled by ERE. Mouse uterine weight test showed that compared with the control group, the aqueous extracts ofC. Semen can increase the uterus index of premature female mice (P<0.01). It can significantly promote the proliferation of MCF-7 cells in the medium without estrogen (P<0.01). The reporter gene controlled by ERE technology showed thatwhen mediated by ERα or ERβ respectively, the normalized luciferase activity of aqueous extracts ofC. Semen was significantly higher than activity of the control group (P< 0.05 orP< 0.01). It was concluded that the aqueous extracts ofC.Semen can increase the uterus index of premature female mice and promote the proliferation of MCF-7 cells in the medium without estrogen. We found the estrogenic effects ofC. Semen for the first time. And the estrogenic effects ofC. Semen were mainly mediated by ERβ.