To observe the effects of hip replacement early postoperative cognitive dysfunction by dexmedetomidine / 中国生化药物杂志

Objective To investigate the effects of hip replacement early postoperative cognitive dysfunction by dexmedetomidine.Methods73 were admitted hip arthroplasty patients from January 2015 to February 2016,randomized, double-blind divided into observation group of 36 cases and a control group of 37 cases.Observe patients before anesthesia intravenous infusion dexmedetomidine given treatment, while the control group were given normal saline infusion, patients received general anesthesia program.Mini-Mental State Examination was observed before and after comparison of two groups of patients (MMSE), central nervous system specific proteins at different times (S100β) level, the third day of neuropsychological test function deterioration.ResultsThe patients in the observation group third and seventh days MMSE scores were higher, compared to the difference between the two groups was significant (P<0.05);and the third day after the seventh day S100β level observation group patients They were lower than the control group, compared to the difference between the two groups was significant (P<0.05);the third day POCD postoperative patients in the observation group was 13.89%, significantly lower than the control group, 48.72% (P<0.05);③ observed after patients total effective rate was 92.86%, significantly higher than 35.14% (P<0.05).ConclusionDexmedetomidine given to patients after hip replacement early cognitive function have a more significant improvement, it can reduce the incidence of POCD.

The effect of hemodialysis filtration combined with blood perfusion in the treatment of patients with uremic encephalopathy / 中国基层医药

Objective To explore the effect of hemodialysis filtration and blood perfusion in the treatment of patients with uremic encephalopathy.Methods 30 patients with uremic encephalopathy were selected,they were treated with hemodialysis filtration and blood perfusion.After the treatment,the treatment effect and postoperative recovery of patients were compared.Results In the treatment,1 case coma for 5 days,after 2 times treatment with death;1 case coma for 7 days.After 3 times the combined treatment did not restore sanity;12 patients of combined treatment after 1 time of the symptoms disappeared completely;After 3 times of treatment on the patients with residual symptoms have less.Consciousness of patients was 25 cases (83.3%).Before treatment,BUN was (28.4 ± 3.1)mmol/L,Scr was (749 ±99.6)mol/L,PTH was (245.7 ±35.2)pg/g,K + was (4.7 ±1.4)mmol/L,Na + was (137.7 ±18.4)mmol/L,Cl -was (90.8 ±16.7)mmol/L,β2 -MG was (25.4 ±3.4)mg/L.After treatment,BUN was (9.1 ±2.4)mmol/L,Scr was (199.7 ±99.2)μmol/L,PTH was (105.6 ±33.6)pg/g,K + was (4.3 ± 1.3)mmol/L,Na + was (135.7 ±14.8)mmol/L,Cl - was (85.0 ±5.2)mmol/L,β2 -MG was (13.4 ±3.1)mg/L. The differences were statistically significant (all P <0.05).Conclusion In the treatment of patients with uremic encephalopathy,hemodialysis filtration and blood perfusion have good effect,should be used in clinical promotion.

The effect of tripterygium glycosides and prednisone in the treatment of adult patients with purpura nephritis / 中国基层医药

Objective To study the effect of tripterygium glycosides combined with prednisone in the treat-ment of adult patients with purpura nephritis.Methods 20 adult patients with purpura nephritis were selected and randomly divided to two groups according to the random number table method.10 cases of the control group were administered with prednisone,10 cases of the observation group were administered with tripterygium glycosides and prednisone.After treatment,the treatment effect and postoperative recovery of the two groups were compared.Results The markedly effective,effective,ineffective,the total effective rates of the observation group were 5 cases(50%), 40 cases(40%),1 case (10%),9 cases (90%),respectively,which of the control group were 3 cases (30%), 4 cases(40%),3 cases(30%),7 cases(70%),there was statistically significant difference in the total effective rate between the two groups(χ2 =4.32,P <0.05);The disappeared time of negative urinary protein,hematuria,edema disappeared,negative hypertension in the observation group were (31.5 ±5.6)d,(35.6 ±6.6)d,(29.6 ±6.4)d, (30.3 ±6.5)d,respectively,which of the control group were (37.5 ±6.5)d,(42.6 ±8.2)d,(35.6 ±7.5)d, (37.7 ±7.3)d,the differences were statistically significant between the two groups(t =2.57,2.62,2.60,2.81,all P <0.05 ).The gastrointestinal discomfort,decreased white blood cells,abnormal liver function,total incidence of adverse reaction rates of the observation group were 1 case(10%),1 case(10%),1 case(10%),3 cases(30%), which of the control group were 1 case(10%),1 case(10%),0 case(0%),2 cases(20%),there was no statistically significant difference in the total incidence of adverse reaction rate between the two groups(χ2 =1.13,P <0.05). Conclusion In the treatment of adult patients with purpura nephritis,tripterygium glycosides combined with predni-sone has good effect,and can improve the symptoms of patients required a shorter time.

Fusion to Mistic improved fatty acid desaturase Fat-1 overexpression in E .coli / 重庆医学

Objective To construct a highly efficient expression plasmid of eukaryotic nuclear membrane protein Omega 3 fatty acid desaturase gene Fat‐1 in E .coli .Methods Using molecular cloning technology to construct the recombinant prokaryotic expression plasmid pET32a Fat‐1 and pET32a‐Mistic‐Fat‐1 fused with Membrane proteins expression chaperon mistic ;the two re‐combinant plasmids were transformed into E .coli strain BL21 (DE3) ,the expression of Fat‐1 protein and M110 Fat‐1 protein in‐duced by IPTG were identified by SDS‐PAGE and gray degree analysed the amount of expression ,further identified by Western blot .Results The results of enzyme digestion and sequencing demonstrated that we successfully constructed the prokaryotic ex‐pression vectors pET32a Fat‐1 and pET32a‐Mistic‐Fat‐1;SDS‐PAGE and Western blot showed that Fat‐1 fatty acid desaturase wasn′t significantly induced ,but the overexpression of M110 Fat‐1 fusion protein was obtained in E .coli ,accounting for 15% of the total amount of whole cell proteins .Conclusion The fusion with Mistic proteins to express the Fat‐1 gene has realized the overex‐pression of eukaryotic nuclear membrane integrated protein Omega 3 fatty acid desaturase in prokaryotic host .

Association of the Serotonin Receptor 3E Gene as a Functional Variant in the MicroRNA-510 Target Site with Diarrhea Predominant Irritable Bowel Syndrome in Chinese Women

BACKGROUND/AIMS: The functional variant (rs56109847) in the 3'-untranslated regions (3'-UTR) of the serotonin receptor 3E (HTR3E) gene is associated with female diarrhea predominant irritable bowel syndrome (IBS-D) in British populations. However, the relationship of the polymorphism both to HTR3E expression in the intestine and to the occurrence of Chinese functional gastrointestinal disorders has yet to be examined. METHODS: Polymerase chain reaction amplification and restriction fragment length polymorphism analyses were employed to detect polymorphisms among Chinese Han women, particularly 107 patients with IBS-D, 99 patients with functional dyspepsia (FD), 115 patients with mixed IBS and 69 patients with IBS-D + FD. We also assessed microRNA-510 (miR-510) and HTR3E expression in human colonic mucosal tissues with immunohistochemistry and other methods. Dual-luciferase reporter assays were conducted to examine the binding ability of miR-510 and HTR3E 3'-UTR. RESULTS: Genotyping data showed the variant rs56109847 was significantly associated with IBS-D, but not with FD, mixed-IBS, or FD + IBS-D. HTR3E was abundantly expressed around the colonic mucosal glands but less expressed in the stroma. miR-510 expression decreased, whereas HTR3E expression increased in the colonic mucosal tissue of patients with IBS-D compared with those in controls. HTR3E expression was significantly higher in patients with the GA genotype than that in patients with the GG genotype. The single-nucleotide polymorphisms disrupted the binding site of miR-510 and significantly upregulated luciferase expression in HEK293 and HT-29 cells. CONCLUSIONS: The single-nucleotide polymorphisms rs56109847 led to reduced microRNA binding and overexpression of the target gene in intestinal cells, thereby increasing IBS-D risk in the Chinese Han population. The decreased expression of miR-510 might contribute to IBS-D.

Neuronal differentiation of cell subsets with stem cell characteristics in adult rat meningeal tissues / 中国组织工程研究

BACKGROUND:Neural stem cels have the potential to differentiate into neurons and glial cels to replace the injured brain cels, so as to achieve the purpose of repairing nerve injury. OBJECTIVE:To observe the neuronal differentiation ability of cel subsets with stem cel characteristics in the adult rat meningeal tissues. METHODS:Under anesthesia, the meningeal tissues were obtained from adult Sprague-Dawley rats to make cel suspension folowed by inoculation and subculture. Then, the Nestin immunofluorescence staining was performed. The third generation cels were culturedin vitro with complete culture medium containing trichostatin A. After 7 days of induction, western blot assay was used to detect the expression of NF-200 and BM88 proteins in neural cels. RESULTS AND CONCLUSION: At 24 hours of culture, some spherical cels were suspended and some cels adherent. In addition, some spherical cels scattered gradualy formed the clone spheres, and the growth rate decreased with the increasing volume. The positive expression of Nestin was detected by immunocytochemistry staining, and the cel nucleus was stained blue by Hoechst staining. BM88 and NF-200 proteins were al expressed at 7 days of neural induction. These findings indicate that the cel subsets with stem cel characteristics in the adult rat meningeal tissues can differentiate into neurons after in vitro induction.

The construction and the expression of V5 epitope fused human androgen receptor vector in the yeast cell / 生物医学工程学杂志

When we try to establish the gene recombinant yeast cell to screen the androgenic endocrine disruptors, the key procedure is the androgen receptor (AR) expression in the yeast cell. For this purpose, we obtained the GPD (glyceraldehyde-3-phosphote dehydrogenase) promoter from the yeast genosome of W303-1A using PCR system and inserting it into Swa I and BamH I sites of pYestrp2. The new constructed vector was named pGPD. The V5 epitope tag DNA with a 5'-BamH I and a 3'-EcoR I sticky end was cloned into the corresponding site of the pGPD vector to yield the vector of pGPDV5. The 2 723 bp full length AR ORF amplified by PCR from pcDNA3.1/AR was fused to V5 epitope tag DNA in pGPDV5 to give the AR yeast expression vector of pGPDV5/AR. This fused vector was transformed into the yeast cell (W303-1A). Western blot was used to detect the V5 fused protein of AR, in the protocol of which the primary monoclonal antibody (IgG(2a)) of mouse anti-V5 and the polyclonal secondary antibody of goat anti-mouse (IgG) linked to horseradish peroxidase (HRP) were used to detect the specific protein in the given sample of the transformed yeast extract. The result showed that the fused protein of AR was expressed successfully in the yeast cell.

Isolation and identification of Metarhizium from Citrus grandis 'tomentosa' GAP base / 中国中药杂志

A type of entomopathogenic fungus of soil in Citrus grandis 'tomentosa' production base was isolated and identified with morphological and molecular biological methods, including pathogenesis, spore characteristic and ITS sequence analysis were conducted. The results showed that eighteen entomopathogenic fungi strains were isolated from the Tenebrio molitor infected in the soil samples, which were identified as Metarhizium anisopliae var. anisopliae. Based on results above, we concluded that there was quantity of Metarhizium resources in this area. These provided the useful information for controlling some pests of C. grandis by using these strains of fungus.

The effects of TATA-box in CYC1 promoter on the reporter gene regulated by ERE in the recombinant yeast cell / 生物医学工程学杂志

The complementary oligonucleotides, each with two consensus estrogen response element (ERE)-sequences and 5'-Hind III and 3'-Sph I sticky ends were artificially synthesized. A solution with both the complementary DNA sequences was heated to 95'C and cooled down to room temperature to form double strand DNA (dsDNA). The set was cloned into the corresponding sites of CYC1 promoter of the pERE-CYC-yEGFP to yield pERE-CYCalpha-yEGFP vector. The two different reporter vectors, pERE-CYC-yEGFP and pERE-CYCalpha-yEGFP, the 2ERE, were placed in the CYC1 promoter. The former promoter downstream ERE contains alpha and beta-TATA boxes and the latter has only alpha-TATA box. The two different reporter vectors were transformed into the yeast cells that express human estrogen receptor alpha (ERalpha). Incubation of the recombinant yeasts with the six estrogenic compounds for 4 hours showed that the recombinant cell containing pERE-CYCalpha-yEGFP would give very poor dose-response curves, in contrast to the recombinant cell containing pERE-CYC-yEGFP which produced well-shaped dose-response curves. So it is necessary for this bioassay that alpha and beta-TATA boxes in the minimal CYC1 promoter when the promoter is used as a rapid and high throughput system for screening estrogenic chemical products.

UPLC-Q-TOF/MS analysis of naringin and naringenin and its metabolites in rat urine and feces after intragastric administration of alcohol extract of Exocarpium Citri grandis / 药学学报

To analyze naringin, naringenin and its metabolites in rat urine and feces after intragastric administration of alcohol extract of Exocarpium Citri Grandis, healthy SD rats were fed with alcohol extract of Exocarpium Citri Grandis for 3 days. On the last day, 0-24 h feces and 0-4 h, 4-8 h, 8-24 h urine were collected and analyzed by UPLC-Q-TOF/MS. The post-acquisition data were processed using Metabolynx The result is that naringin and its 6 metabolites, naringenin and its 4 metabolites were detected in the urine of rat. Meanwhile, naringin and its 3 metabolites, naringenin and its 2 metabolites were detected in the feces of rat.

UPLC-Q-TOF-MS analysis of naringin and naringenin and its metabolites in rat plasma after intragastrical administration of alcohol extract of exocarpium Citri grandis / 中国中药杂志

<p><b>OBJECTIVE</b>To analyze naringin, naringenin and its metabolites in rat plasma after intragastric administration of exocarpium Citri grandis alcohol extract.</p><p><b>METHOD</b>Rat blood samples were collected 1.0 hour after oral administration of 50 g x kg(-1) exocarpium Citri grandis alcohol extract and analyzed by UPLC-Q-TOF with MS(E) function. The post-acquisition data were processed using Metabolynx.</p><p><b>RESULT</b>Naringin (M1), naringenin (M2), naringin-5-O-glucuronide (M3), naringin-4-O-glucuronide (M4), glucuronide conjugate of naringenin (M5), naringin-4-O-sulfate (M6), methylated conjugate of hydroxylated naringenin (M7), glucuronide and sulfate conjugate of naringenin (M8), glucuronide conjugate of hydroxylated naringenin (M9) in rat plasma were detected. M3, M4, M6 were first reported as the metabolites of naringin. M7, M9 were first reported as the metabolites of naringenin.</p><p><b>CONCLUSION</b>The results indicated that naringin, naringenin can be metabolited as the forms of glucuronidation, sulfation and naringenin can also be metabolited as the forms of methylation with hydroxylation and glucuronidation with hydroxylation in vivo after administration.</p>

Establishment of Transgenic cell model based on anti-oxidative response element / 生物医学工程学杂志

To establish a transgenic cell model based on anti-oxidative response element (ARE) and green fluorescence protein(GFP) reporter gene, the TK minimal promoter was amplified by PCR and cloned into pEGFP-N1 for constructing reporter vector pTK-GFP/Neo. Four synthetic oligonucleotide ARE motifs were annealed and purified and then were inserted into pTK-GFP/Neo one by one to construct the eukaryotic reporter vector p4ARE-TK-GFP/Neo. Two reconstruct eukaryotic reporter vectors were transfected into HepG2 cells mediated by lipofectamine. The positive clones were obtained by the screen of G418. The cell model was tested with PDTC and tBHQ, well known inducers of phase II enzymes, by determining GFP activity. The results showed that the expression level of GFP was significantly increased by PDTC and tBHQ, and a transgenic cell model based on ARE was established successfully.

Establishment of reporter gene-based cell screening model for discovering new chemopreventive agents / 中国中药杂志

<p><b>OBJECTIVE</b>To discover new chemopreventive agents, a drug screening cell model based on reporter gene and antioxidative response element (ARE) has been established.</p><p><b>METHOD</b>Four repeas of ARE DNA binding conserved sequences were synthesized and cloned into a GFP expression vector. This construct was stably transfected into HepG2 cells in vitro. The cell model was tested with known chemopreventive agents and the effects of resveratrol, protocatechuic aldehyde, ursolic acid and oleanolic acid at different concentration (0, 12.5, 25, 50, 100, 200 micromol x L(-1)) were observed by determining reporter gene GFP activity.</p><p><b>RESULT</b>The induce level of GFP was regulated by ARE and the dose-dependence in a certain range was observed. The induce level of GFP by resveratrol was significantly increased.</p><p><b>CONCLUSION</b>The method can be used to screening of chemopreventive agents from Chinese traditional medicine by measurement of luminescent value of expressed GFP in wells of microtiter plate.</p>

Study on purification and immunologic effect of 42KD protein from group B Streptococcus / 中华微生物学和免疫学杂志

Objective To look for a vaccine on the base of surface protein which can elicit protective immunity.Methods Purification of 42KD protein from Streptococcus serotype Ⅲ was used one-step chromatography.Antibody response was evaluated in mice immunized by 42KD protein with different immunization dose and by the different immunization procedure.After immunizing the mice in the optimum procedure,we challenge the mice in intradermal with Streptococcus to observe the active and passive protection.Results The molecular weight(Mr)of the protein was 42.33×103 and its point isoelectric(pI)was 6.6.Its property might be a glycoprotein which could be released to supernatant.The highest titer of antibody against 42KD protein Was 1:24576.Which indicated that 42KD protein had good immunogenicity.The immunity exponent of NIH mice was 100 in test group and infant mice was 40.Conclusion The data suggests that 42KD protein may elicit protective immunity to adult and infant mice.Through protecting test with different immunizing doses,the findings indicate that protecting capability is relative to the titer of IgG specificantibody.

Bioassay of recombinant green fluorescent protein gene yeast cell for a high throughput to screen estrogenic compounds / 生物工程学报

We developed the recombinant green fluorescent protein gene yeast cell to screen estrogenic compounds based on two episomal vectors. In the expression vector the expression of human estrogen receptor alpha(hERalpha) was driven by 3-glyceraldehydephosphate dehydrogenase (GPD) promoter; in the reporter vector the expression of the yeast enhanced green fluorescent protein (yEGFP) gene was under the control of the estrogen response element (ERE). The vectors were transformed into yeast cell (W303-1A) to construct GFP recombinant yeast cell. Incubation of the yeast cell with various concentrations of the estrogenic compounds led to expression of the reporter gene product GFP in a dose dependent manner. Compared to other yeast bioassays, the yeast cell for environmental estrogen bioassay based on yEGFP reporter gene did not need cell wall disruption or the addition of a substrate or reagent. This yEGFP assay was performed completely in 96 well plates. So this test system can be used as a rapid and high throughput system for screening estrogenic chemical products, which has the characteristics of the sensitivity, reproducibility and cheapness.

Monitoring of Electrophile Chemical Pollutants Using Transgenic Firefly Luciferase Gene Cells / 环境与健康杂志

Objective To assess quickly the potential environmental pollution by using the luciferase transgenic cells. Methods Many electrophile compounds in the environment can generate oxidative stress,so that the transcription of certain protective genes is induced via specific DNA motifs called electrophile response elements (EPREs). We have made a vector containing a single EPRE fused to the TK minimal promoter and the gene encoding firefly luciferase (EPRE-LUC) by adopting bio-molecular techniques. From this vector the stable LUC expression vector regulated by EPRE had been successfully reconstructed. This reporter construct was transfected into HeLa cells,and the clones resistant to G418 were selected. The resistant cells were treated by the different concentrations of sodium arsenate(NaAsO2),cadmium chloride (CdCl2),mercury chloride(HgCl2) and diethyl mateate (DEM). After that,the expression of luciferase was determined by luciferase assay kit. Results The correct construct frame of LUC reporter vector regulated by EPRE was identified by DNA sequencing; the dose-dependent relationships between the LUC expression and the test substance concentration were found. Among them,the relationship produced by DEM was the most significant. Conclusion The LUC transgenic cells regulated by EPRE have been successfully constructed.

Assessment of the Quality of Two Different Species of Exocarpium Citri Grandis / 广州中医药大学学报

[Objective] To assess the quality of Citrus grandis Osbeck. Var. tomentosa Hort. (CGO Var. TH) and Citrus grandis (L. ) Osbeck (CGO). [Methods] The macroscopic feature and microscopic structure of CGO Var. TH and CGO were compared. Their content of total flavones (TF) was determined by spectrometry, naringin content and fingerprint by high performance liquid chromatography (HPLC) and the content of rhoifolin by thin-layer chromatography (TLC). Their expectorart, anti-inflammatory and antitussive effects were observed by phenol red test, auricular swelling test and ammonia-water-induced cough method. [Results] Obvious differences such as villi in the exocarp and non-grandular hair in the stem existed in the macroscopic feature and microscopic structure of CGO Var. TH and CGO. The contents o{ total flavones and naringin in CGO Var. TH were markedly higher than that in CGO and the rhoifolin content in CGO Var. TH was ten times as much as that in CGO. Markedly difference was also found in their fingerprint. The expectorant, anti-inflammatory and antitussive effects in CGO Var. TH were better than those in CGO. [Conclusion] The quality of CGO Var. TH is much better than that of CGO.

Determination of the Rhoifolin in Exocarpium Citri Grandis By RP-HPLC / 中药新药与临床药理

Objective To establish a method for the determination of Rhoifolin content in Exocarpium citri grandis (ECG). Methods The RP-HPLC method was operated on MACHEREY-NAGEL 100-5 C_(18), with methanol and wateracetic acid (61: 4) as mobile phase, gradient elution, flow rate at 1.0 mL/min, and detection wavelength at 345nm. Results In the range from 0.816 ?g to 13. 056?g, Rhoifolin is in a good linearity with chromatographic peaks area, r=0.9995, the average recovery ratio is 101.064%, RSD=3.10%(n=5). The content of Rhoifolin in Tomentulosus Exocarpium Citri Grandis(TECG) ranged from 1.33% to 0.177% and that in Glabrous Exocarpium Citri Grandis (GECG) is less than 0.17%. Conclusions RP-HPLC method is with accuracy and a good reproducibility for the determination of the Rhoifolin content in ECG; the difference of Rhoifolin content in TECG and GECG is significant.

Studies on Genuineness of Exocarpium Citri Grandis / 广州中医药大学学报

Objective To explore the genuineness of Exocarpium Citri Grandis(ECG) from Huazhou city of Guangdong province.Methods We used the method of high performance liquid chromatography to detect the naringin content in ECG from different producing areas of Huazhou city.Random amplified polymorphic DNA analysis was used for the examination of genetic distance,and plasma-atomic emission spectrometry for the detection of soil elemental abundance of 8 elements such as aluminium(Al),kalium(K),calcium(Ca),ferrum(Fe),titanium(Ti),boron(B),magnesium(Mg),and manganese(Ma).The correlation of the above three parameters was analyzed by statistical software SPSS 11.5.Results Ca abundance in the surface soil layer had an obvious effect on the content of naringin,and the difference of Al and K abundance in subsoil layer was correlated with the genetic distance of ECG.Conclusion The genuineness of ECG is probably related with the abundance of phlopopitum in the soil of producing areas of Huazhou city.

Development of a Rapid Screening System of Environmental Estrogen Based on Recombinant Yeast Cell / 环境与健康杂志

Objective To develop a rapid screening method of environmental estrogen in vitro. Methods Estrogen responsive element (ERE) was inserted into the upstream of Lac Z (?-galactosidase ) reporter gene and the reporter gene was regulated under estrogen receptor(ER) in the recombinant yeast cells by the molecular biological technique. When the yeast cells was affected by environmental estrogen, conformational change of ER lead to expression through binding to ERE. There exist the functional relationships between the expression and the pollution degree of environmental estrogen. Therefore, the pollution degree can be reflected by assaying the activity of ?-galactosidase. Results It was identified by PCR that recombinant yeast cell, whose Lac Z reporter gene regulated under estrogen receptor, was successfully reconstructed. The experimental results showed that the time-dose-effect response had no significant changes when the recombinant yeast cell bad been exposed to ?-estradiol for 4 and 8 hours respectively; among the three methods of yeast disintegration, the ultrasonic vibration method had more advantage compared with that of the other methods and this dose-effect curve exhibited "S" shape; among 3 test chemical products, ?-estradiol was the most estrogenic activity, biosphenol A take second place, estradiol benzoate was the lowest. Conclusion The assay system of yeast cell for environmental estrogen hormone mediated by estrogen receptor is stable and applicable.