Functional Identification of a Multi-product Sesquiterpene Synthase from Ganoderma sinense / 中国实验方剂学杂志

Objective: Ganoderma sinense is one of the most famous medicinal fungi in China. Because it is a model medicinal fungus of basidiomycete,the functional identification of its sesquiterpene synthase is of great significance for the biosynthesis and regulation studies of fungal sesquiterpene. Method: A sesquiterpene synthase gene was discovered by mining the genome data of G. sinense. The sesquiterpene's conservative motifs was analyzed through multiple sequence alignment with two identified sesquiterpene synthases of G. sinense and three terpenoid synthases in the Nr database,which have the highest similarity to it. Subsequently,heterologous expression was observed in Escherichia coli,and protein expression was detected by SDS-PAGE. Volatile compounds were collected by solid phage microextraction (SPME) and detected by GC-MS. Result: The enzyme containing sesquiterpene conserved motifs DDXXDE and NSE/DTE were efficiently expressed in E. coli,and 11 sesquiterpenes were synthesized with endogenous FPP as the substrate. The product α-cadinol at 18.6 min was considered to be the main product,and previous studies showed a significant anticancer activity. According to the comparison with chemical standards,three products were identified as γ-muurlene,α-muurolene and δ-cadinene,respectively. Conclusion: The functional identification of multi-product sesquiterpene synthase from G. sinense can promote the study on the mechanism underlying its product diversity,and lay a foundation for the production of rare or novel sesquiterpenes by improving the catalytic activity of enzymes with enzyme engineering technology.

Bioactivity and Chemical Constituents of Different Polar Parts from Blueberry Leaves / 中国药学杂志

OBJECTIVE: To study the bioactivity and chemical constituents of different polar parts from blueberry leaves. METHODS: Blueberry leaves were extracted by ethanol and then the extract was sequentially partitioned into five fractions. Silicagel and Sephadex LH-20 chromatographic methods were applied to isolate and purify compounds. Their structures were elucidated by physiochemical properties and spectral analysis.The DPPH• radical scavenging activity, α-glycosidase and pancreatic lipase inhibition activity of different polar parts and partial compounds were determined. RESULTS: The n-butyl alcohol fraction(BF) showed the highest DPPH• radical scavenging activity and α-glycosidase inhibition activity. The ethyl acetate fraction(EAF) showed the strongest pancreatic lipase inhibition activity. A total of five compounds were isolated from the EAF, and their structures were identified as β-sitosterol(1), quercetin-3-O-α-L-arabinofuranoside(2), quercetin(3), quercetin-3-O-β-D-glucopyranoside(4) and 1-O-caffeoylquinic acid(5). A total of two compounds were isolated from the BF, and their structures were identified as quercetin-3-O-α-L-arabinoside(6) and quercetin-3-O-β-D-glucuronide(7). The results showed that compounds 3 and 5 had very good DPPH• radical scavenging and pancreatic lipase inhibitory activity, and compounds 1 and 3 had good α-glucosidase inhibitory activity. CONCLUSION: The different polar parts and compounds of blueberry leaves show strong DPPH• radical scavenging activity, α-glycosidase and pancreatic lipase inhibition activity. Compounds 1, 2, 5 and 6 are isolated from blueberry leaves for the first time.

Efficacy of real-time PCR for detecting Clostridium difficile infection: comparison with enzyme-linked fluorescent spectroscopy-based approaches / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the diagnostic efficacy of real?time polymerase chain reaction (q?PCR) for Clostridium difficile infection (CDI) in comparison with routine culture and enzyme?linked fluorescent spectroscopy?based aprroaches.</p><p><b>METHODS</b>Stool samples were collected from suspected CDI cases in General Hospital of Guangzhou Military Command of PLA between May and December in 2016. All the samples were examined with 3 methods, namely enzyme?linked fluorescent spectroscopy for detecting Clostridium difficile toxin A/B (CDAB), detection of glutamate dehydrogenase (GDH), and q?PCR for amplification of Clostridium difficile?specific gene tpi and toxin gene (tcdA/tcdB), with the results of fecal culture as the reference for evaluating the diagnostic efficacy of the 3 methods.</p><p><b>RESULTS</b>Of the total of 70 fecal samples, 13 (18.57%) were found to be positive for Clostridium difficile, including toxin?producing strains in 6 (8.57%) samples. The sensitivity, specificity, positive predictive value, negative predictive value and diagnostic coincidence rate of q?PCR for tpi were 92.31%, 91.23%, 70.59%, 98.11% and 91.43%, respectively, which were significantly higher than those of GDH test (84.62%, 84.21%, 55.00%, 96.00%, and 84.29%, respectively; Χ=24.881, P<0.001). The sensitivity of q?PCR for tcdA/cdB was significantly higher than that of enzyme?linked fluorescent spectroscopy for CDAB in detecting CDI (66.67% vs 33.33%; Χ=35.918, P<0.001).</p><p><b>CONCLUSION</b>Both CDAB detection and q?PCR have a high specificity in detecting CDI, but GDH detection has a good sensitivity, and all these 3 methods have a high negative predictive value. Compared with other detection methods, amplification of tpi and tcdA/tcdB using q?PCR allows more rapid, sensitive and specific detection of CDI.</p>

Application of comfort care for dental out-patients / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore and apply the nursing methods of comfort care for dental out-patients.</p><p><b>METHODS</b>Control group included 103 dental out-patients who were first treated in General Dentistry Department of West China Hospital of Stomatology from June to August 2008. Experimental group included 105 dental out-patients who were first treated in the same hospital from September to November 2008. Conventional nursing methods were used for control group, and comfort care nursing methods were used for experimental group. The patients' degree of satisfaction with nursing and oral health knowledge rate after first treatment and nursing were investigated.</p><p><b>RESULTS</b>The patients' degree of satisfaction with nursing of the control and experimental groups were 72.816% and 98.095%, and the patients' oral health knowledge rate of the two groups were 57.282% and 93.333%. Both of the investigating results had obvious difference.</p><p><b>CONCLUSION</b>Comfort care for dental out-patients can improve patients' degree of satisfaction with nursing and increase patients' oral health knowledge rate. Simultaneously, comfort care can improve the specialized quality of dental nurses and would advantage to build a harmonious relationship between nurses and patients.</p>

Effect of sFRP2 on the biological behavior of HepG2 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of sFRP2 on the biological behavior of human hepatoma carcinoma HepG2 cells.</p><p><b>METHODS</b>HepG2 cells were infected with recombinant adenovirus containing mouse sFRP2 gene, and then the proliferation, cell cycle distribution, expression of tumor metastasis related factors (CD44, CD82/KAI1, EMMPRIN) and beta-catenin protein, and migration ability of the cells were detected by MTT, FCM, immunohistochemistry, Western blot and Transwell inserts, respectively.</p><p><b>RESULTS</b>sFRP2 protein inhibited the proliferation of HepG2 cells, and increased the percentage of G0/G1 period cells. Expression of CD44 and CD82/KAI1 proteins, which could inhibit invasion and metastasis of tumor cells, was upregulated. However, EMMPRIN protein, which could promote the above properties of tumor cells decreased in HepG2 cells infected with the recombinant adenovirus containing mouse sFRP-2 gene. Western blot demonstrated that beta-catenin was expressed in HepG2 cells and there was no significant difference between the treated and the control groups. Transwell insert test showed sFRP2 protein decreased the migration ability of HepG2 cells.</p><p><b>CONCLUSION</b>The recombinant adenovirus containing mouse sFRP-2 gene could infect HepG2 cells. sFRP2 protein could significantly reduce the capability of proliferation, invasion and metastasis of HepG2 cells.</p>

Clinical effects of Fu Fang Ya Tong Ding on gingivitis and pericoronitis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the clinical effects of Fu Fang Ya Tong Ding on treatment of gingivitis and pericoronitis.</p><p><b>METHODS</b>120 clinical patients with gingivitis or pericoronitis were randomly divided into 3 groups (40 patients in each group). After routine rinse treatment for all patients, patients in the test group were treated with Fu Fang Ya Tong Ding, patients in the positive group were treated with iodine glycerol, while that time patients in the negative group received no treatment anymore. Ten minutes after treatment, visual analogue scale (VAS) was used to record the severity of pain for each patient. 3 days and 7 days later, pain and inflammation degree were also recorded by pain three-degree scoring method and index of gingivitis. The total treatment effects were evaluated under a comprehensive clinical treatment standard.</p><p><b>RESULTS</b>10 minutes after treatment, 40.0% of patients in the test group had almost no pain, while no obvious reduction of pain was found in the control group. 3 days, 7 days after the treatment, 92.5%, 95.0% of patients in the test group had no pain, and 55.0%, 90.0% of patients in the positive group had no pain. In the negative group, there were 47.5% of patients which pain was still remained in 7 days. 7 days after treatment, gingival index in the test group reduced by 25.0% and 42.8% compared with the positive and negative groups (P<0.05). 3 days after treatment, 62.5%, 45.0% and 30.0% patients separately in the test, positive and negative groups manifested good effects under the comprehensive clinical treatment standard; after 7 days, 97.5%, 92.5% and 77.5% patients in the 3 groups manifested good effect. The group using Fu Fang Ya Tong Ding had better effects than groups using iodine glycerol or only applying routine rinsing treatment group (P<0.05).</p><p><b>CONCLUSION</b>Fu Fang Ya Tong Ding can treat gingivitis and pericoronitis through significantly reducing inflammation and pain.</p>