1.Gene detection and clinical study of rhinovirus isolated from children with acute low respiratory tract infection in Shanghai
Jun LI ; Qi-Rong ZHU ; Hui YU ; Xin-Huan GU ;
Chinese Journal of Infectious Diseases 2007;0(12):-
Objective To understand human rhinovirus (HRV) etiology of acute lower respiratory tract infection (ALRTI) in children in Shanghai area and establish a nested reverse transcription- polymerase chain reaction (nested RT-PCR) assay.Methods Three hundred and forty-two naso- pharyngeal secretion (NPS) samples from ALRTI cases who were hospitalized were collected during January 2005—December 2005.Nested RT-PCR techniques were used to detect HRV-specific RNA.The PCR products were sequenced and data of nucleotides were analyzed.The proportion of HRV infection in children with ALRTI,the distribution of gender,age and season,and clinical char- acteristics were also investigated.Results Forty-six (13.5%) of 342 samples were HRV positive detected by nested RT-PCR.The sequences of 15 positive samples shared high homology of 83%- 97% with HRV sequence in GenBank.Within the 15 positive samples,nucleotide homology varied from 64.4% to 98.4%,and the ratio of genetic variation was from 1.6% to 48.3%./00.These 15 ampli- cons attribute to the two branches of HRV cladogram.The sequences of 15 amplieons were highly varied,in which single nucleotide mutation and several nearby nueleotides mutations were found. Ribonucleotide deletion and insertion in the nucleotide sequence was also found.HRV positive sam- ples were detected in 33 boys and 13 girls,respectively.The ratio of infection cases between boys and girls was 2.5:1.Of 46 HRV infected cases,27 (58.7%) were less than 12 months of age and 38 (82.6%) were less than 3 years old.HRV infected ALRTI occured all the year round and peaked from March to May.Of the patients whose NPS samples were HRV positive detected by nested RT-PCR,45 patients were diagnosed with bronchopneumonia and 1 was diagnosed with asthmatic bronchitis.Fever of most patients was moderate.The peripheral blood leukocyte counts in thirty-nine (84.8%) patients were less than 10?10~9/L.Neutrophil percentages in thirty-seven (80.4%) patients were less than 0.50.C-reactive protein of thirty-six (78.3%) patients were less than 8 mg/L. All of these features were the characteristics of viral pneumonia.The complications were not common and conditions of most patients were not severe.All the children were cured.Conclusions This nes- ted RT-PCR technique is highly specific,rapid and convenient for the detection of HRV RNA in NPS of patients with ALRTI and the genome of HRV viruses is highly variable.The incidence of HRV infection predominates in children in Shanghai area.ALRTI of HRV is short of specificity and condi- tions of most patients are not severe and their prognoses are fine.
2.Nursing of patients with nasal facial soft tissue defect treated with transplantation of free foot skin flap
Lili ZHU ; Chunli ZOU ; Rong SHI ; Huan JIANG ; Xiling TANG
Modern Clinical Nursing 2014;(12):29-31,32
ObjectiveTo summarize the nursing measures for patients with nasal facial soft tissue defect treated with transplantation of free foot skin flap.MethodThe clinical data of 13 patients with transplantation were reviewed to summarize the nursing measures.ResultAll skin flaps of the 13 patients survived.ConclusionEnough preoperative preparation,active psychological nursing and careful postoperative observation are critical for the success of the operation.
3.Preliminary analysis of bitter substances in spica of Prunella vulgaris.
Xin ZHAI ; Meng-Qian XI ; Qiao-Sheng GUO ; Huan-Huan HAN ; Xiang ZHANG ; Wei YANG ; Rong-bo ZHENG ; Xiao-Dan HUANG ; Huan-Rong ZHU
China Journal of Chinese Materia Medica 2014;39(3):423-426
Volatile oil components and the contents and types of amino acid in spica of Prunella vulgaris were analysed by GC-MS and amino acid analyzer. Esters, fatty acids, aromatic hydrocarbon, ketone and several alcohol compounds were identified by mass spectrum comparison. In these ingredients, beta-ionone smelled aroma of cedar, raspberry, nerolidol showed weak sweet soft orange blossom flavor, neroli tasted sweet and fresh, nerolidol tasted sweet with light aroma of wood, hexadecanal showed a weak aroma of flowers and wax, alpha-sinensal had rich and fresh sweet orange flavor. To some extent, these types of aromatic substances can affect the taste of herbal tea or decoction made of Spica Prunellae. Among amino acids detected, natural amino acids accounted for a larger proportion, and those natural amino acids showed bitterness, slight bitterness, sourness (freshness), sweetness, slight sweetness, sourness (slight freshness). The results indicated that bitter and slightly bitter amino acids have the greatest impacts on the sense of Spica Prunellae.
Amino Acids
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analysis
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Gas Chromatography-Mass Spectrometry
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Oils, Volatile
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analysis
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Prunella
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chemistry
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Taste
4.Fluorescence labeling for human bone marrow mesenchymal stem cells with PKH26
Xing-Zhong WANG ; Wen-Rong XU ; Wei ZHU ; Huan YANG ; Chun QIAO ; Hui QIAN ; Jia-Bo HU ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
Objective To establish a method of labeling human mesenchymal stem cells (MSCs) with PKH26 in vitro.Methods MSCs were cultured and labeled with PKH26 according to the manufacturer's instruction.The growth,fluorescence intensity and serial subcuhivation of labeled MSCs were analyzed with the confocal laser microscope and the flow cytometry.The biological characteristics of labeled MSCs were investigated by RT-PCR.Results The labeled MSCs appeared red fluorescence and the labeling rate was 100 percent.During serial subcuhivation of labeled MSC from passage 1 to passage 7,the fluorescence intensity and the labeling rate of MSCs were gradually decreased.The biological features such as morphology,growth,expression level of nucleostemin and GAPDH gene and capability of differentiation into osteoblast in vitro were not affected by labeling.Conclusion Labeling the human MSCs with PKH26 is an effective and practical method,which can be used as an important tool in the study on the homing, plasticity and transplantation of MSCs.
5.Adipose-derived stem cell transplantation on motor function of ischemic rats
Xue-Qian YUAN ; Li-Feng ZHANG ; Huan-Rong WANG ; Tai-Qing ZHU
Chinese Journal of Neuromedicine 2010;09(10):1005-1008
Objective To observe the survival, migration and differentiation of adipose-derived adult stem (ADAS) cells and the recovery of motor function of ischemic rats after ADAS cell transplantation. Methods Eighteen adult male SD rats, weighted about 250-300 g, were chosen and received left middle cerebral artery occlusion (MCAO) operation. And then, they were equally randomized into untreated group, control group and ADAS cell transplantation group. Tail vein injection of Dulbecco's modified eagles medium (DMEM) and ADAS cells were performed in the control group and ADAS cell transplantation group 3 h after MCAO, respectively. These animals were euthanized 14 d after MCAO. Immunofluorescence for BrdU, NSE, MAP-2 and GFAP were processed to identify the survival, migration and differentiation of grafted ADAS cells in the brain, and at the same time, scores of neurological deficit scale were used to assess the improvement of motor function. Results After the transplantation, numerous ADAS cells labeled with BrdU were presented in the ischemic points and surrounding areas. A few BrdU/GFAP, BrdU/NSE and BrdU/MAP-2 -positive cells were noted in the ischemic points of ADAS transplantation group 14 d after MCAO. The neurological functional recovery in the ADAS cell transplantation group was significantly improved as compared with that in the control group 14 d after MCAO (P<0.05). Conclusion ADAS cells can migrate into the ischemic hemisphere and differentiate into neuron-like and astrocytic-like cells after the injection by venous approach in the rat models with MCAO. The intravenous administration of ADAS cells into rats with MCAO leads to good functional outcome and few lesion sizes.
6.Meta-analysis on acupuncture and moxibustion for treatment of ulcerative colitis.
Jing-ping MU ; Huan-gan WU ; Zhi-quan ZHANG ; Hui-rong LIU ; Yi ZHU ; Zheng SHI ; Xiao-mei WANG
Chinese Acupuncture & Moxibustion 2007;27(9):687-690
OBJECTIVETo assess the effectiveness and safety of acupuncture and moxibustion for treatment of ulcerative colitis.
METHODSRandomized controlled trials or clinical controlled trials of acupuncture and moxibustion for interfere of ulcerative colitis in recent 10 years were reviewed and Meta-analysis was made for the literature results.
RESULTSAltogether 11 papers of clinical study were enrolled. Heterogeneous tests were conducted for the results of the 11 studies, as a result, chi2 = 8.55, P = 0.67. The fixed effect model was used for statistical analysis, after combination OR = 3.82, confidence interval of 95% was 2.65-5.52. The rhombus was located at the right side of the medium line. After Z test, Z = 7.14, P < 0.01, the therapeutic effect and the cured rate in the treatment group were significantly higher than those of the control group.
CONCLUSIONThe therapeutic effect of acupuncture and moxibustion on ulcerative colitis is superior to that of western medicine with safety and less adverse reactions.
Acupuncture Therapy ; Colitis, Ulcerative ; therapy ; Humans ; Moxibustion ; Randomized Controlled Trials as Topic
7.Protective effect of the extract of Terminalia catappa leaves on acute liver injury induced by D-GalN in mice.
Xin-hui TANG ; Jing GAO ; Huan DOU ; Yan-ping WANG ; Li-zhi XU ; Zeng-rong ZHU ; Qiang XU
China Journal of Chinese Materia Medica 2004;29(11):1069-1073
OBJECTIVETo study the hepatoprotective effect of the extract of Terminala catappa leaves (TCE) and the possible mechanisms underlying its protection on acute liver injury induced by D-Galactosamine (D-GalN).
METHODIn vivo: D-GalN-induced liver injury model was used to evaluate the effect of TCE on the activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in mice. Structure of liver was observed and liver mitochondrial swelling was measured following D-GalN injection without or with TCE. In vitro: D-GalN-induced primary cultured hepatocytes injury model was used to value the effect of TCE on cultured hepatocytes. Cell viability was measured by means of MTT assay, and the AST and superoxide dismutase (SOD) activities in supernatant of cultured cells were investigated also.
RESULTIn acute hepatic injury test, with oral pretreatment of TCE, remarkable rises in serum AST and ALT activities (2.95 fold and 3.35 fold) induced by D-GalN were obviously reversed and significant morphological changes were remarkably lessened. In addition, the decrease in sensitivity of mitochondrial swelling to the exotic Ca2+ stimulation induced by D-GalN was also prevented by TCE. In primary cultured hepatocytes of mice, it was found that incubation with TCE could prevent the decrease in cell viability in a dose-dependent manner. It was also found that both the increase in AST level (1.9 fold) and the decrease in SOD activity (48.0%) in supernatant of primary cultured hepatocytes induced by D-GalN could be inhibited by pretreatment of TCE.
CONCLUSIONTCE has hepatoprotective activity and the mechanisms underlying its protective effect may be related to its antioxidant activity and protection on both hepatocytes and liver mitochondria.
Animals ; Cells, Cultured ; Chemical and Drug Induced Liver Injury ; blood ; etiology ; pathology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Galactosamine ; Liver ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Pregnancy ; Protective Agents ; pharmacology ; Terminalia ; chemistry
8.Effect of allogeneic blood transfusion and preoperative autologous blood donation on perioperative immune function in patients with gastrointestinal tumor
Xun ZHOU ; Huan WANG ; Xiaoqian LIU ; Nana ZHU ; Rong XIA ; Jianrong GUO
The Journal of Clinical Anesthesiology 2018;34(2):144-148
Objective To investigate the effect of allogeneic transfusion and preoperative autologous blood donation on perioperative cellular immunity and humoral immunity in patients with gastrointestinal neoplasms.Methods Sixty gastrointestinal cancer patients undergoing radical surgery,33 males and 27 females,aged 53 69 years,weighing 47-70 kg,ASA physical status Ⅰ or Ⅱ,were included in this study.Blood transfusion was started when the blood loss was more than 200~400 ml,Hb<70 g/L,and the patients were randomly divided into the preoperative autologous blood donation group (group P,n =30):intraoperative blood transfusion using the stored autologous blood transfusion;allogeneic blood transfusion group (group A,n =30):allogeneic blood transfusion.The levels of T lymphocyte subsets,NK cells and IL-2,IL-10,TNF-α,perforin (PF) concentration and plasma immunoglobulin IgG,IgA and IgM levels.Results The percentage of CD3+,CD4+,NK cells and the ratio of CD4+/CD8+ in group A at the end of surgery to 7 d after operation were significantly lower than those at the time of admission (P<0.05).The percentage of CD3+,CD4+ (P< 0.05).The level of IL-2 in group A was significantly lower than that in group P (P<0.05) 1-7 d after operation,and the level of IL-10 in group A was significantly higher than that in group P (P< 0.05).The levels of IgG and IgA 3 d after operation in group A were significantly lower than those in group P (P<0.05).The levels of IgG and IgA in group P were significantly decreased at the end of operation and recovered to preoperative levels 1-3 d after operation (P < 0.05).Conclusion Allogeneic blood transfusion can reduce the percentage of T-cell subsets and NK cells in cancer patients and delay their recovery,and also can transiently reduce the content of immunoglobulin IgG and IgA in plasma and thus affect the immune function of patients.However,the preoperative autologous blood donation has a slight effect on postoperative immune function in cancer patients.
9.Relationship between cytokine gene polymorphism and susceptibility to hepatitis B virus intrauterine infection.
Qi-rong ZHU ; Shao-qing GU ; Hui YU ; Jian-she WANG ; Xin-huan GU ; Zuo-quan DONG ; Lin-e FEI
Chinese Journal of Epidemiology 2005;26(4):236-239
OBJECTIVETo explore the possible relationship between cytokines (TNF-alpha, IFN-gamma, IL-4 and IL-10), which were expressed abnormal quantity in the peripheral blood to intrauterine HBV infectious children, gene single nucleotide polymorphism (SNP) and susceptibility to HBV intrauterine infection.
METHODSA cross sectional study on molecular epidemiology was carried out. The subjects were selected from outpatients of the hepatitis B vaccine special clinics of our hospital. According to intrant criteria, children under high risk of HBV intrauterine infection were divided into immuno-failure group (group I) and immuno-effective group (group II) while children without high risk were included in the control group. Four gene SNP sites of TNF-alpha-238, IFN-gamma + 874, IL-4-590 and IL-10-1082 region were determined by real-time quantitative fluorescent PCR.
RESULTSSignificant differences of TNF-alpha-238 A allele frequency were found between group I and group II (chi(2) = 6.797, P < 0.05) as well as between group I and control group (chi(2) = 9.513, P < 0.05). No evident difference of TNF-alpha-238 A was found between group II and control group (chi(2) = 0.047, P > 0.05). Significant differences of IFN-gamma + 874 A allele frequency were found between group I and group II (chi(2) = 7.238, P < 0.05), and between group I and the controls (chi(2) = 5.199, P < 0.05) but no significant difference was found between group II and control group (chi(2) = 0.602, P > 0.05). Significant differences of IL-4-590 C/T allele frequency were not found between group I and group II (chi(2) = 0.632, P > 0.05), group I and control group (chi(2) = 0.584, P > 0.05), or between group II and control group (chi(2) = 0.004, P > 0.05) respectively. Significant differences of IL-10-1082 G allele frequency were found between group II and group I (chi(2) = 10.359, P < 0.001), and between group II and the controls (chi(2) = 35.418, P < 0.001), but not found between group I and control group (chi(2) = 1.759, P > 0.05).
CONCLUSIONThis study suggested the possibility that TNF-alpha-238 A allele and IFN-gamma + 874 A allele were associated with HBV intrauterine infection. There was no evident relationship between IL-4-590 C/T allele SNP and susceptibility to HBV intrauterine infection, but the IL-10-1082 G allele seemed to be associated with preventive efficacy to HBV intrauterine infection.
Case-Control Studies ; Child ; Child, Preschool ; Cytokines ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Hepatitis B ; transmission ; Humans ; Infant ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Interferon-gamma ; genetics ; Interleukin-4 ; genetics ; Polymorphism, Genetic ; Pregnancy ; Pregnancy Complications, Infectious ; Retrospective Studies ; Risk Factors ; Tumor Necrosis Factor-alpha ; genetics
10.Application of fluorescent real-time polymerase chain reaction in analyzing the epidemic of influenza among children in Guangzhou area in 2006.
Yi CHEN ; Rong ZHOU ; Bing ZHU ; Ming-qi ZHAO ; Ru-xu CHANG ; Ying-ying ZHANG ; Mi-si XIAO ; Huan-hui CHEN ; Qi-yi ZENG
Chinese Journal of Pediatrics 2008;46(8):613-617
OBJECTIVETo investigate the prevalence of influenza virus infections in children in 2006 using the real-time PCR method.
METHODS(1) Consulting the most conserved sequence NP gene of influenza virus, after comparing with the NP gene sequences of influenza virus in GenBank, one pair of specific primers and one TaqMan probe were designed for each subtype of influenza virus by the software Primer Express. The sensitivity of influenza was evaluated by testing known positive samples which had been two-fold diluted. The specificity of real-time PCR for influenza virus detection was assessed by cross testing 60 isolates of influenza A, 16 isolates of influenza B, and by testing a variety of other respiratory viruses positive samples; (2) 281 nasopharyngeal aspirate samples were detected by real-time PCR and virus isolation; (3) the 12 301 specimens from the patients of Guangzhou Children's Hospital were tested by using the real-time PCR method. Furthermore, the real-time PCR reagent was evaluated by comparing with the result of virus isolation.
RESULTS(1) The sensitivity of real-time PCR developed in this study for influenza A detection was 1:2(22) and for influenza B was 1:2(20) in two-fold serially diluted way. (2) No positive results were found in cross testing of other viruses positive specimens. (3) Influenza virus was detected from 1687 cases (13.71%) out of the 12 301 cases, including 773 cases (45.8%) positive for subtype A and 914 cases (54.2%) positive for subtype B; 455 out of 525 (86.7%) of influenza B positive specimens and 70 out of 525 (13.3%) of influenza A (H1N1) positive specimens were from patients seen during January to April; 419 out of 1118 (37.5%) specimens positive for influenza B and 699 out of 1118 (62.5%) specimens positive for influenza A (H1N1) were from patients seen from May to August. Influenza virus could be identified from 1380 samples by the methods of virus isolation, accounting for 81.80% of the 1687 positive samples detected by real-time PCR. All the influenza virus subtype A was H1N1.
CONCLUSIONThe real-time PCR method developed in this study was sensitive and specific for detecting influenza A and B in clinical specimens. During 2006, influenza A and influenza B co-circulated. The predominant virus was influenza B from January to April, peaking in April. Influenza A (H1N1) prevailed from May to August, with the peak in June.
Child ; China ; epidemiology ; Epidemics ; Humans ; Influenza A Virus, H1N1 Subtype ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Polymerase Chain Reaction ; methods ; Prevalence ; RNA, Viral ; isolation & purification ; Sensitivity and Specificity