1.Eukaryotic expression and identification of human DC-SIGN and EGFP fusion protein
Lulu XU ; Shuliang GUO ; Yongai LUO
Chinese Journal of Immunology 1985;0(05):-
Objective:To construct the eukaryotic vector of human DC-SIGN and EGFP fusion protein,and to identify the protein in cell line COS7.Methods:RT-PCR,T-vector and pEGFP-C1 vector were used to construct the recombinant expressing plasmid encoding for the fusion protein of DC-SIGN and EGFP.COS7 cells were transfected with the plasmid.Real-time PCR and laser scanning confocal microscope were used to quantificate expression of the fusion protein and cell function of uptaking BCG.Results:DC-SIGN cDNA was successfully cloned into the eukaryotic vector pEGFP-C1.The recombinant vector was transfected into COS7,real-time PCR test showed the amount of mRNA encoding for the fusion protein was 4.52?1011 copies/ml and laser scanning confocal microscope confirmed that the cells could uptake BCG.Conclusion:We have constructed a recombinant vector expressing DC-SIGN and EGFP fusion protein.
2.Meta-analysis of javanica oil emulsion injection combined with radiotherapy in the treatment of esophageal carcinoma
Jian CHEN ; Lulu LI ; Dingru ZHANG ; Yufei GUO ; Xincheng GUO
The Journal of Practical Medicine 2015;(13):2182-2185
Objective To evaluate the advantage and clinical value of javanica oil emulsion injection combined with radiotherapy for esophageal carcinoma. Method Randomized controlled trials (RCTs) concerning javanica oil emulsion injection combined with radiotherapy for esophageal carcinoma were made on electronic databases of CBM, CNKI, Wangfang, VIP and PubMed from 1990 to February 28, 2015. Quality of the included RCTs was assessed by Jadad scoring , and Meta-analyses were performed by RevMan5.3 software. Results Seven RCTs involving 575 patients were included in Meta-analysis. All studies were in low quality. The results of Meta-analyses showed that Javanica oil emulsion injection combined with radiotherapy for esophageal carcinoma, compared with control groups, could increase the recent curative effect (P < 0.000 01) and improve the quality of live (P = 0.000 8), decrease the hematologic toxicities (P = 0.03) and the incidence rate of radioaction esophagitis (P = 0.02), but not enough evidence was found to prove that it can enhance survival rate (P = 0.25). Conclusions The results of Meta-analysis indicate that javanica oil emulsion injection may have therapeutic effects on the treatment of esophageal carcinoma. Yet the effects of javanica oil emulsion injection still need to be confirmed by large multi-center randomized controlled trials.
3.Effects of berberine on mRNA expression of 11β-Hydroxysteroid dehydrogenase type Ⅰ in insulin-resistant HepG2 cells
Xin NIE ; Lulu CHEN ; Kunquan GUO ; Xiaozheng WANG
Journal of Chinese Physician 2012;14(7):874-877,880
Objective To investigate the effect of berberine on glucose absorption and mRNA expression of 11 β-hydroxysteroid dehydrogenase type 1 ( 11β-HSD1 ) in insulin-resistant HepG2 cell model.Methods HepG2 cells were incubated with high-concentration insulin for 24 hours to build insulin-resistant cell model.In order to evaluate the cells for insulin resistance,the cells were cultured with different concentrations of insulin for 24 hours.The insulin-resistant cells were treated with different concentrations of berberine and insulin for 24 hours and the non insulin-resistant cells were used as a control.The glucose concentration in culture medium was detected by the method of glucose oxidase-peroxidase (GOD-POD).According to the glucose concentrations in blank medium and those in the medium of culturing cells 24 hours later,the rate of absorpting glucose by the cells was calculated.The mRNA expression of 11β-HSD1 in the insulin-resistant cells was detected by the reverse transcription polymerase chain reaction (RT-PCR).Results Incubated with 10-7mol/L insulin for 24 h,the insulin-resistant cell model had been built.The rate of glucose absorption of the model cell treated with high concentration berberine ( 10 μmol/L) was significantly improved [(42.53 ±1.99)% vs (28.16±1.99)%,t =12.9457,P <0.01].High-concentration berberine showed a strong synergy with insulin on glucose absorption of the model cells.As the cells became resistant to insulin,the mRNA expression of 11β-HSD1 increased significantly compared to non insulin-resistant cells( relative expression quantity was (4.60 ±0.96 vs 0.67 ±0.42,t =4.9476,P <0.05 ).While the mRNA expression of 11β-HSD1 reduced in the insulin-resistant cells after treated with high-concentration berberine,and the relative expression quantity was not significantly different with non insulin-resistant cells ( 1.12 ±0.35 vs 0.67 ±0.42,P >0.05).However,low-concentration ( 1 μmol/L) of berberine had not the same role.Conclusions It is concluded that one of the acting mechanism of berberine improving the insulin sensitivity may be that the mRNA expression of 11β-HSD1 is downregulated in the insulin-resist-ant liver cell model depending on concentration.
4.Transportation and Absorption of Thermo-sensitive Gel Mediated 5-FU Multiple Emulsion in Caco-2 Cell Monolayer Model
Lulu WANG ; Huihui GUO ; Wensheng ZHENG ; Shaohua CHEN ; Yanxing HAN
China Pharmacist 2016;19(9):1641-1643,1655
Objective: To develop 5-FU multiple emulsion entrapped into thermo-sensitive gel (5-FU-DEG) and detect the ab-sorption and transportation in Caco-2 cell monolayer model. Methods:The 5-FU multiple emulsion was prepared by a two-step emulsif-ying method. Poloxamer 407 (P407) was used as the thermo-sensitive material and sodium alginate (SA) was used as the bioadhesive material for the preparation of 5-FU-DEG. Caco-2 cell monolayer model was used to investigate the transportation and absorption of 5-FU. Results:5-FU-DEG gelled at the ambient temperature and turned into liquid below 10℃ The apparent permeability coefficient (Papp) of 5-FU-DEG was 1.47 ±0.11 ×10 -5(cm·s-1), which was about 6 times higher than that of 5-FU water solution(2.39 ± 0.21 ×10 -6 cm·s-1)(P<0.01). The cellular uptake rate of 5-FU-DEG was (17.1 ±0.24) %, which was 3.9 times greater than that of 5-FU water solution (4. 41 ± 0. 23%)(P<0. 01). Conclusion:5-FU-DEG can efficiently enhance the transportation and ab-sorption of drug in rectal site by using micro-emulsion technology combined with thermo-sensitive technology, which can be an effective rectal delivery system for 5-FU to treat rectal cancer.
5.Correlation between self-efficacy and care ability of stroke patients′family caregivers
Lulu LV ; Hong GUO ; Liyun HU ; Qingxia LIU ; Xiaojing XU
Modern Clinical Nursing 2016;15(8):6-10
Objective To investigate the current status of self-efficacy and care ability among stroke patients′ family caregivers and explore the correlation between them. Method A total of 79 stroke patients and family caregivers were engaged in the investigation by demographical data questionnaires, general self-efficacy scale and questionnaire for assessing the comprehensive care-giving ability of stroke caregivers. Results The score on self-efficacy of the family caregiver was (27.16 ± 5.64) and the total score on care ability was (90.01 ± 16.57). There was a positive correlation between self-efficacy and care ability of family caregivers(P<0.05). Conclusions The self-efficacy and care ability of stroke patient′s family caregivers are at middle level. Their self-efficacy is positively correlated with their care ability. The nurses can improve the ability of daily caring of family caregivers by enhancing their self-efficacy.
6.Laboratory evaluation and field trial of activation indigenous microbial displacements in the reservoirs after polymer flooding.
Jianjun LE ; Lulu BAI ; Rui WANG ; Menghua GUO ; Jiyuan ZHANG ; Zhaowei HOU ; Xiaolin WU
Chinese Journal of Biotechnology 2015;31(7):1129-1138
Most main oilfields in China have already entered a "double high" development stage (high water cut, high recovery degree). To further enhance oil recovery in reservoirs after polymer flooding (RAPFs), an efficient activator formulation for promoting metabolism of endogenous microorganism was studied by aerogenic experiments, physical simulation experiments, electron microscopy scanning and pyrophosphate sequencing. Results show that the activator could activate the endogenous microorganisms in the injected water and make the pressurized gas reach 2 MPa after 60 d static culture of the activator in a high pressure vessel. The oil recovery efficiency of natural core physical simulation flooding can be improved by more than 3.0% (OOIP) in RAPFs when injected 0.35 PV activator with 1.8% mass concentration, and a lot of growth and reproduction of activated endogenous microorganism in the core was observed by electron microscopy scanning. Field trial with 1 injector and 4 producers was carried out in the east of south II block of Sa Nan in December 2011. By monitoring four effective production wells, changes of carbon isotope δ13C (PDB) content of methane and carbon dioxide were -45 per thousand to -54 per thousand and 7 per thousand to 12 per thousand. Compared with east II of Sa Nan block, the oil amount increased by 35.9%, water cut stabled at 94%. The incremental oil was 5 957 t during the three and a half years, which provides an alternative approach for further improving oil recovery in similar reservoirs.
Carbon Dioxide
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chemistry
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Carbon Isotopes
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analysis
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China
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Diphosphates
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chemistry
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Methane
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chemistry
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Oil and Gas Fields
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microbiology
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Polymers
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Water
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Water Microbiology
7.Origin and Phylogenetic Characteristics of Dengue Virus Isolated from the Large Outbreak of Dengue in Guangdong Province in 2014
Qianfang GUO ; Guohui CUI ; Danyun FANG ; Huijun YAN ; Junmei ZHOU ; Lulu SI ; De WU ; Lifang JIANG
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(1):21-28
[Objects]To isolate and identify the pathogen of the large outbreak of dengue in Guangdong province in 2014. To understand the origin and the phylogenetic characteristics of the isolates ,and provide scientific foundation for the surveillance and prevention of dengue fever.[Methods]Collected the patient serum samples over all the Guangdong province during the 2014 outbreakperiod,isolated and identified the virus from these samples. Amplified complete E gene and complete genome with certain primers and sequenced all the products. Then the Phylogenetic ,Bayesian phylogeography and mutations analysis were carried.[Results]40 DENV-1 strains were isolated and identified. 40 complete E gene sequences and 6 complete genome sequences of DENV-1 were obtained. Phylogenetic analysis with E gene sequences revealed that the 40 isolates were classified into two genotypes including 16 genotypeⅠ(Asia)and 24 genotypeⅤ(America/Africa). 14 genotypeⅠisolates were clustered closest with isolates from Guangdong province(2013)and Sigapore(2013)which share the nucletide identities of 99.6% ~ 99.9%,other two genotypeⅠisolates were clustered with strains from Malaysia (2013) and both share the nucletide identities of 99.7%;24 genotypeⅤisolates were all classified in one clade with striains from Bangladesh(2009),China(2009)and Bhutan(2013)which share nucletide identities of 99.0%-99.9%. Further analysis with six complete genome sequences showed that five isolates were clustered closest with strains isolated from Guangdong province(2013)share the nucletide identities of 99.6%-99.8% while the sixth stains closest with strains isolated from Myanmar(2002)share the nucletide identities of 98.8%. The isolates have five amino acid mutations compared with strains epidemic in Guangdong province in 2013,three mutations(S88V,E203G,T275R)are in the EⅡdomain and one mutation (S305P)is in the EⅢdomain which associated with virulence.[Conclusions]During the outbreak in Guangdong province in 2014, DENV-1 is the predominant causative serotype,and there are at least two different kinds of genotypes of DENV-1 largely epidemiced in the whole province. Evolution analysis reveals the multiple origins of the isolates which may origin from Guangdong province , Sigapore,Malaysia,Myanmar so that we should enhance the study and surveillance of autochthonous and vectors in order to understand the epidemic way of dengue in Guangdong province. The isolates have had four mutations in the domain associated with virulence which remain further study to know their biological effects.
8.The gene frequencies of HLA-Cw in Guangdong Han population and its significance
Lulu XIAO ; Hongjing MA ; Xiaolin YIN ; Xin YE ; Weidong ZHANG ; Kunyuan GUO
Chinese Journal of Immunology 1986;0(04):-
Objective:To explore the gene frequencies of HLA-Cw and to analyze the recognition characteristic between HLA-Cw and KIR in Guangdong Han population.Methods:An auto semi-quantitative PCR-RSSO method was adopted to detect the HLA-Cw genotypes of a sample of 122 bone marrow donors.Results:The gene frequencies of HLA-CW 03,07,01,08,04,14,15,12,06,05,16 were 0.237 1 , 0.215 9 , 0.175 2 , 0.112 9 , 0.050 5 , 0.041 9 , 0.041 9 , 0.037 6 , 0.033 3 , 0.008 2 , 0.004 1 respectively.HLA-Cw 02,04,05,06 belong to “group 1” recognizing KIR-2DL1/2DS1.HLA-Cw 01,03,07,08 belong to “group 2” recognizing KIR-2DL2/2DL3;2DS2/2DS3.There was a significant difference between the two groups in total HLA-Cw gene frequencies in Guangdong Han population( P
9.Establishment of a miniature pig model of mandibular osteoradionecrosis.
Song LI ; Dianji FANG ; Zhanyi WANG ; Lulu ZHAI ; Zhaorong NING ; Yanwei GUO
West China Journal of Stomatology 2015;33(6):570-574
OBJECTIVETo establish a miniature pig model of mandibular osteoradionecrosis (ORN).
METHODSA single dose of irradiation (IR25 Gy, 28 Gy) was delivered via an electronic linear accelerator to the right mandible of 24 miniature pigs by utilizing three dimensional conformal radiotherapy (3D-CRT). The first molar of the mandible was extracted 2 months after radiation. Mandibular ORN was diagnosed through gross observation, X-ray film, CT, and histopathologic examination.
RESULTSAt 3-4 months after radiation, all animals suffered from mandibular ORN after dental extraction. The symptoms of ORN were more severe in the 28 Gy group than in the 25 Gy group.
CONCLUSIONA miniature pig model of mandibular ORN can be established through 25 Gy irradiation + dental extraction.
Animals ; Mandible ; Mandibular Diseases ; pathology ; Molar ; Osteoradionecrosis ; pathology ; Particle Accelerators ; Radiation Injuries, Experimental ; pathology ; Radiotherapy, Conformal ; adverse effects ; Swine ; Swine, Miniature ; Tooth Extraction
10.Therapeutic effect of curcumin and hippocampal brain-derived neurotrophic factor on a rat model of Alzheimer’s disease
Xiaoying GENG ; Aixiang ZHANG ; Jian QIAN ; Gang GUO ; Lulu ZHANG ; Jing WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;(2):273-278,297
ABSTRACT:Objective To study the effect of curcumin on the learning and memory ability in a rat model of Alzheimer’s disease (AD).Methods AD rat model was prepared using intraventricular injection of Aβ1-42. Curcumin was acutely (single injection before the behavioral tests)or chronically (injected for 6 consecutive days) injected intraperitoneally at doses of 50,100 or 300 mg/kg.Brain-derived neurotrophic factor (BDNF)protein (1 μg/side)or BDNF shRNA (2×10 5 units/side)was infused into the hippocampus.The behavioral changes in Y-maze,open field test and Morris water maze and the expression of BDNF in the hippocampus were analyzed. Results Acute treatment with curcumin had no significant effects on the spontaneous alteration,locomotor activity or water maze latency of AD rats.AD rats treated chronically with curcumin (300 mg/kg ) showed significant elevation in the spontaneous alternation (P <0.000 1)in Y-maze and memory ability in the water maze test (P <0.05 )compared with those in the saline group.Chronic treatment with 100 and 300 mg/kg of curcumin induced an increased level of BDNF in the hippocampus as compared with the saline controls (P <0.05 and <0.000 1). Intrahippocampal injection of BDNF significantly decreased the escape latency of AD rats in the water maze (F 4,2 9 5=5.813,P <0.01 ).Rats chronically injected with curcumin combined with shBDNF showed no difference in the swimming time in Ⅱ quadrant as compared with saline controls (P =0.657).However,rats in 100 mg/kg curcumin group,BDNF group and sham group had significantly increased swimming time than the saline controls (P <0.05, P <0.05 and P <0.000 1,respectively).Conclusion Curcumin may activate the downstream signaling pathways by upregulating the expression of BDNF and ultimately contribute to the improvement of learning and memory in AD rats.