2.Effect of Jianpi Bushen Qingchang Huashi Recipe on Proliferation of Bone Marrow Mesenchymal Stem Cells.
Lei ZHU ; Hong SHEN ; Li LIU ; Pei-qing GU ; Jia-fei CHENG ; Lu ZHANG
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(2):191-195
OBJECTIVETo observe the effect of Jianpi Bushen Qingchang Huashi Recipe (JBQHR) on proliferation and migration of bone marrow mesenchymal stem cells (BMSCs).
METHODSBMSCs were isolated and cultured in vitro with adherence screening method to prepare cell suspension. No drug intervention was given to BMSCs in the vehicle control group. JBQHR at 0.39, 0.78, 1.56 µg/mL was added in BMSCs of low, mid, and high dose JBQHR groups for co-incubation. Its effect on the proliferation of BMSCs was detected by CCK-8. BMSCs migration and chemotactic ability was detected using Transwell method. Each dose JBQHR combined ERK kinase inhibitor U0126 was set up as control. The phosphorylation of extracellular regulated protein kinase (ERK) and CAMP responsive element-binding protein (CREB) were detected by Western blot.
RESULTSCompared with the vehicle control group, the proliferation of BMSCs and BMSCs migration number could be promoted in the 3 JBQHR groups (P < 0.05). Besides, the proliferation of BMSCs was better in mid and high dose JBQHR groups than in the low dose JBQHR group (P < 0.05). Compared with the vehicle control group, the phosphorylation of ERK and CREB could be elevated in the 3 JBQHR groups (P < 0.05), and could be inhibited by U0126 (P < 0.01). Compared with the low dose JBQHR group, the phosphorylation of ERK increased in mid and high dose JBQHR groups with statistical difference (P < 0.05).
CONCLUSIONJBQHR could promote the proliferation and migration of BMSCs, and its mechanism might be related to ERK/CREB signaling pathway
Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclic AMP Response Element-Binding Protein ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; MAP Kinase Signaling System ; Mesenchymal Stromal Cells ; cytology ; drug effects
3.Extraction of DNA Using Monoclonal Anti-DNA and Colloidal-gold Beads
Jie-Ping PEI ; Jun HE ; Hua TAN ; Hong ZHU ; Qing DUAN ;
Microbiology 1992;0(02):-
To developing a simple, rapid and sensitive immunoaffinity method for extracting DNA before amplification by the polymerase chain reaction(PCR). Using monoclonal anti-DNA bound to colloidal-gold beads(which called immunocolloidal-gold ) and extracting DNA from the specimen before amplification by PCR. The immunoaffinity method we use is extremely efficient in extracting DNA at low concentration, also, the beads carrying the DNA/anti-DNA complexes can be added directly to a PCR mixture without elution, and the presence of PCR inhibitors in specimens has no effect on amplification. When applied to serial dilution bacteria suspension, the detection of limit can be 100 CFU/mL, and the sensitivity of detecting artificial soil sample and milk sample were 101 and 100 CFU/mL. The immunoaffinity method described here can sever as a sensitive and practicable method for extracting DNA , particularly where samples have low concentrations of DNA or where the material is in poor condition.
4.Experimental study of amniotic lacrimal duct stent used to prevent dry eye of castrated rabbits
Mingyang MA ; Qing YUAN ; Qi LIU ; Kangcheng LIU ; Peiwen ZHU ; Honghua KANG ; Nan JIANG ; Lei YE ; Chonggang PEI ; Yi SHAO
Recent Advances in Ophthalmology 2017;37(8):709-713
Objective To explore the effects of amniotic lacrimal duct stenting on the prevention of dry eye in castrated rabbits.Methods Thirtysix healthy male rabbits were selected,the third eyelid were cut off and antiinfection treatment were given,which were randomly divided into 3 groups (12 cases in each group),the castrated male rabbits models were made.Among them,group A was negative control group,group B was dry eye model group,group C was group of lacrimal amniotic membrane group.At 2 weeks before implantation of amniotic lacrimal duct stent,2 weeks,4 weeks and 6 weeks after implantation,the fluorescent (FL) examination,Western blot,Schirmer I examination,immunofluorescence staining and corneal confocal microscopy were performed.Results The levels of tear secretion and FL in the three groups among different time points were significantly different (F=7.126,P =0.009;F =9.658,P =0.016),and there were significant differences among three groups (F =12.582,P =0.005;F =13.187,P =0.013).The tendency of tear secretion and FL in the three groups were also significantly changed (F =8.531,P =0.007;F =10.652,P =0.019).The epithelial basal cells at 6 weeks after implantation in three groups were 3811 ±414,3820 ± 314,2789 ± 353,and the density of inflammatory cells was 266 ±28,266 ± 29,67 ± 13,there were significant differences among three groups (F =13.442,P =0.012;F =9.231,P =0.021).The K1 6 staining in the duct epithelium were negative,and the expression of α-SMA in the lacrimal duct tissue of group A,B and C was not changed at all time points after implantation of amniotic lacrimal stent,and there was no significant difference (F =14.681,P =0.002).Conclusion The amniotic lacrimal stent implantation has certain effect on the prevention of dry eye in rabbit.
5.Aberrant neuronal expression of mitotic protein, tau and Bax in the rat brain after injection of Abeta(25-35) into the amygdala.
Dao-Jun HONG ; Ai-Lin PEI ; Feng-Yan SUN ; Cui-Qing ZHU
Acta Physiologica Sinica 2003;55(2):142-146
Recent evidence indicates that the aberrant neuronal expression of mitotic proteins in Alzheimer's disease (AD) brain may be related to AD pathological changes. To investigate whether the toxicity of beta-amyloid protein (Abeta) induces mitotic proteins expression in adult rat brain, we used immunohistochemical and integral optical density analytic method to analyze the adult rat brains, which had been injected with Abeta(25-35) into unilateral amygdala. Results showed that the levels of neurofibrillary tangle (NFT) related phosphorylated tau protein and apoptosis related protein Bax were increased in Abeta(25-35) injected rat brains, meanwhile the aberrantly expression of mitotic protein cyclin A and cyclin B1 was also detected at 7 d after operation, but the level of cyclin A decreased and cyclin B1 disappeared at 21 d. Immunofluorescence double labeling presented that cyclin B1 was partially co-localized with Bax or phosphorylated tau protein, whereas Bax and phosphorylated tau protein seldom co-localized. These results suggest that Abeta causes mitotic protein expression in adult brain neurons, which may die through apoptosis or may be affected by AD NFT-related tau phosphorylation.
Alzheimer Disease
;
metabolism
;
physiopathology
;
Amygdala
;
drug effects
;
metabolism
;
physiopathology
;
Amyloid beta-Peptides
;
toxicity
;
Animals
;
Cyclin A
;
metabolism
;
Cyclin B1
;
metabolism
;
Male
;
Neurons
;
metabolism
;
Peptide Fragments
;
toxicity
;
Phosphorylation
;
Random Allocation
;
Rats
;
Rats, Sprague-Dawley
;
bcl-2-Associated X Protein
;
metabolism
;
tau Proteins
;
metabolism
6.Analysis of polymorphisms in the promoter region of interleukin-1beta by restriction fragment length polymorphism-PCR.
Ai-Qing WEN ; Jun WANG ; Kai FENG ; Pei-Fang ZHU ; Jian-Xin JIANG
Chinese Journal of Traumatology 2004;7(5):271-274
OBJECTIVETo investigate the frequencies of -1470, -511 and -31 single nucleotide polymorphisms (SNPs) in the promoter of IL-1beta and its haplotype constitution in Chongqing population.
METHODSOne hundred and twelve healthy Chongqing people were enrolled in this study. Polymorphisms at -1470 (G to C), -511 (T to C) and -31 (C to T) of IL-1beta were genotyped with the method of restriction fragment length polymorphism (RFLP). Haplotype frequencies were analyzed by Arlequine software.
RESULTSFrequencies of IL-1beta -1470, -511 and -31 SNPs were 41.67%, 50% and 45.33%, respectively. Genotype frequencies of -1470 locus were 39.81%, 37.04% and 23.15% for G/G, G/C and C/C respectively. As for T-511C SNP, genotype frequencies of T/T, T/C and C/C were 29.91%, 40.18% and 29.91%, respectively. Genotyping results of C/C, C/T, and T/T of -31 locus were 35.51%, 38.32% and 26.71% respectively. Haplotype analysis found that there were mainly three haplotypes constituted by three SNPs, ie., G-T-C, C-T-C and G-C-T.
CONCLUSIONSPolymorphisms exist in the promoter of IL-1beta in Chongqing population. Three SNPs locate in the same haplotype block.
Asian Continental Ancestry Group ; genetics ; China ; Cohort Studies ; Female ; Genetics, Population ; Genotype ; Haplotypes ; Humans ; Interleukin-1 ; genetics ; Male ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational ; Promoter Regions, Genetic ; Sensitivity and Specificity
7.Linkage disequilibrium analysis of -31, -511, -1470 single nucleotide polymorphisms in the promoter of IL-1 beta in Chongqing population.
Ai-qing WEN ; Jian-xin JIANG ; Pei-fang ZHU ; Kai FENG
Chinese Journal of Medical Genetics 2004;21(4):412-414
OBJECTIVETo investigate the frequencies of -31, -511 and -1470 single nucleotide polymorphisms (SNPs) in the promoter of IL-1 beta in Chongqing population and address the question whether they are in linkage disequilibrium (LD).
METHODSOne hundred and twelve healthy Chongqing people were enrolled in this study. Polymorphisms at -31 (C>T), -511 (T>C) and -1470 (G>C) of IL-1 beta were genotyped with the method of restriction fragment length polymorphism (RFLP). Haplotype frequencies were evaluated with Arlequine software. Two-point LD analyses were done with the software TransposerV1-0. The P values were determined by Pearson chi square test analysis.
RESULTSAllelic frequencies of IL-1 beta -31, -511 and -1470 were 45.33%, 50.00% and 41.67%, respectively. The dominant haplotypes comprising the three loci were T-C-G (44.1%), C-T-C(40.3%) and C-T-G(8.8%), LD analyses revealed that none of the LD parameters(delta value) was 0. Meanwhile, chi square test showed P<0.005.
CONCLUSION-31, -511 and -1470 loci in the promoter region of IL-1 beta are in strong linkage disequilibrium. And this study provides a basis for searching disease-related IL-1 beta haplotye.
Adult ; China ; Female ; Gene Frequency ; Genotype ; Haplotypes ; Humans ; Interleukin-1beta ; genetics ; Linkage Disequilibrium ; Male ; Middle Aged ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; genetics
8.Clinical features of pure erythroid leukemia--case report and review of literature.
Yan-Ming ZHANG ; De-Pei WU ; Yu-Mei SUN ; Shu-Hua LU ; Ming-Qing ZHU
Chinese Journal of Hematology 2008;29(5):293-295
OBJECTIVETo report a case of pure erythroid leukemia.
METHODSThe clinical features, treatment and prognosis of a rare case of pure erythroid leukemia were reported, and the related literature was reviewed.
RESULTSThe pure erythroid leukemia patient was diagnosed by 90.4% pronormoblasts in bone marrow, 99.5% for erythroid antigen CD71, 67.4% for glycophorin A were detected, while no differentiation antigen of myeloid, lymphoid and megakaryocyte lineages were observed. HAG (homoharringtonine + Cytarabine and G-CSF) regimen were administered with no effect. The patient developed multiple organ failure and died soon.
CONCLUSIONPure erythroid leukemia has a fulminant clinical course with poor response to chemotherapy and worse prognosis.
Humans ; Leukemia, Erythroblastic, Acute ; therapy ; Male ; Middle Aged ; Prognosis
9.Comparison of 3 microsurgical approaches to the treatment of varicocele: report of 120 cases.
Qing SHI ; Hai ZHU ; Xin-Sheng WANG ; Pei-Tao WANG ; Si-Chuan HOU
National Journal of Andrology 2013;19(10):931-934
OBJECTIVETo compare the outcomes of the three microsurgical strategies, inguinal high ligation (IHL), retroperitoneal high ligation (RHL) and low ligation (LL) of internal spermatic veins, in the treatment of varicocele.
METHODSWe retrospectively analyzed 120 cases of varicocele, which were equally divided into groups I , II and III to be treated by IHL, RHL and LL of internal spermatic veins, respectively. We compared the operation times, post-operative complications, recurrence rates and results of pre- and post-operation semen analysis among the three groups.
RESULTSThe mean operation time was significantly longer in group III ( [55 +/- 6 ] min) than in I ([35 +/- 10] min) and II ([42 +/- 12] min) (P<0.05), while the rate of post-operative complications remarkably higher in group I (4 cases of hydrocele [10% ] and 3 cases of epididymitis [7.5%]) than in II (2 cases of hydrocele [5%] and 2 cases of epididymitis [5%]) and III (1 case of hydrocele [2.5%] and 1 case of epididymitis [2.5%]) (P<0.05). Six to 12 (mean 9) months of follow-up visit found 2 cases (5% ) of recurrence in group I, 1 case (2.5%) in group II and none in group III, with no statistically significant difference among the three groups (P>0.05). At 12 months after surgery, group III showed significantly higher sperm concentration, grade a +b sperm and the sperm motility than the other two (P<0.05), but no significant differences were observed in these parameters among the three groups preoperatively.
CONCLUSIONAs a microsurgical approach to the treatment of varicocele, LL is better than IHL and RHL of internal spermatic veins in improving the seminal parameters of the patients.
Humans ; Ligation ; Male ; Microsurgery ; adverse effects ; methods ; Retrospective Studies ; Treatment Outcome ; Urologic Surgical Procedures, Male ; adverse effects ; methods ; Varicocele ; surgery
10.A preliminary pathological study on human allotransplantation.
Hui-jun WANG ; Yan-qing DING ; Guo-xian PEI ; Li-qiang GU ; Li-jun ZHU
Chinese Journal of Traumatology 2003;6(5):284-287
OBJECTIVETo observe the survival of hand allograft under the state of immunosuppression and the pathological changes of rejection in the recovery process.
METHODSThe biopsies of the skin, nerve, muscle, tendon and bone tissue of hand allografts during different stages from 1 day to 7 months after operation were observed using routine histological technique.
RESULTSNo significant changes due to rejection in skin, nerve, muscle and bone tissue were observed. But different degrees of weak rejective changes were found on the wall of blood vessels; in the muscle and nerve the reactions were markedly stronger than those found in skin tissues.
CONCLUSIONSThe rejection in deep tissues should be monitored in controlling the rejection of hand allograft.
Adult ; Biopsy ; Graft Rejection ; pathology ; Hand Transplantation ; Humans ; Immunosuppression ; Male ; Skin ; immunology ; pathology ; Transplantation, Homologous