Objective To study the effect of cholecalciterol cholesterol emulsion(CCE)in the zymosan(Z)-induced acute hepatic injury. Meth-ods A total of 36 C57 BL/6 mice were randomly divided into 4 groups,namely negative control(CON)group,CCE group,Z group and CCE+Z group,respectively. There were 9 mice in each group. Mice from CON group and Z group were fed with pure water. Mice from CCE group and CCE+Z group were fed with cholecalciterol cholesterol emulsion 20μL dissolved in 200 mL pure water which was kept in darkness. After 14 days, Z group and CCE+Z group were injected with zymosan at a dose of 500 mg/kg. After 18 hours,all the mice in each group were sacrificed. The liver tissues were harvested for histopathological examination. The serum ALT levels were determined. The molecular expression of IL-6 and IL-18 in liv-er tissue of mice were evaluated by Western blotting and real-time quantitative PCR method. Results The results of histopathological examination showed that liver tissue damage in CCE+Z group was lighter than that of Z group ,and heavier than that of the CON group. Compared to the CON group,Z group had the highest serum ALT level,followed by CCE+Z group,while in Z group was significantly lower than that in CON group(all P<0.05). The expression of IL-6 and IL-18 protein and mRNA showed level of Z group was apparently higher than those of CON group and CCE+Z group(all P<0.05). Conclusion Cholecalciterol cholesterol emulsion can play certain protective effect on zymosan-induced liver injury in mice.

WRKY transcription factors are novel transcriptional regulatory factors, which play an important role in regulating plant development, metabolism and other physiological processes. In this study, a new Dendrobium officinale WRKY transcription factor, designated as DoWRKY1 was cloned by using RT-PCR and RACE (GenBank Accession No. KF953910). Bioinformatic analysis demonstrated that, the full-length cDNA of DoWRKY1 was 1,704 bp. And DoWRKY1 contained a 1,629 bp open reading frame (ORF) that encoding a peptide of 542 amino acid residues. The putative DoWRKY1 protein contained two conserved WRKY domains and it belonged to the group I WRKY family protein. Yeast one-hybrid experiment showed that DoWRKY1 had transcriptional activation ability in yeast, and it could activate the expression of downstream report genes (His3 and Ade2). Semi-quantitative RT-PCR experiment showed that DoWRKY1 expressed in roots, stems, leaves and protocorm-like bodies. Real-time qRT-PCR proved that DoWRKY1 could be induced by methyl jasmonate (MeJA) and chitosan (Chitosan), and the expression level of this gene can reach the expression peak at 2 h and 1 h, respectively. These results are useful for further determination of the regulation function of this gene in secondary metabolism of D. officinale.
Cloning, Molecular ; Dendrobium ; genetics ; Gene Expression Regulation, Plant ; Plant Proteins ; genetics ; Transcription Factors ; genetics

Objective To investigate the correlation between vitamin A and surfactant protein (SP)-B, SP-C in human body,and to explore the effects on lung development and pulmonary function of neonates. Methods We collected the blood samples of 170 pregnant women and umbilical cord serum of their neonatal babies. The levels of vitamin A in pregnant women and their neonatal babies,and the levels of SP-B and SP-C in neonatal umbilical cord serum were detected by ELISA. We conducted a follow-up by standard telephone questionnaire,which we concerned was the number of respiratory tract infection within six months,in order to assess the neonatal pulmonary functions. Results (1) There was a positive correlation between the vitamin A levels in neonatal umbilical cord blood and in the blood of pregnant women(r=0. 866,P<0. 05). (2) There was a positive correlation between the vitamin A levels in neonatal umbilical cord blood and the levels of SP-B,SP-C in the blood(r=0. 817,P<0. 05). (3)In the follow-up of 170 cases of infants within six months,three cases with pneumonia hospitalized more than once,but no respiratory distress syndrome hap-pened. Conclusion Vitamin A can be used as an important biological marker to evaluate the neonatal pul-monary maturity. If we detect the vitamin A levels of pregnant women,increase the intake of vitamin A,we can improve the content of SP-B,SP-C,improve the development of neonatal lung function in growth.

BACKGROUNDHuman embryonic stem (HES) cell derived from human blastocyst can be propagated indefinitely in the primitive undifferentiated state while remaining pluripotent. It has exciting potential in human developmental biology, drug discovery, and transplantation medicine. But there are insufficient HES cell lines for further study.

METHODSThree oocyte donors were studied, and 3 in vitro fertilization (IVF) cycles were carried out to get blastocysts for the establishment of HES cell line. Isolated from blastocysts immunosurgically, inner cell mass (ICM) was cultured and propagated on mouse embryonic fibroblasts (MEFs). Once established, morphology, cell surface markers, karyotype and differentiating ability of the cell line were thoroughly analyzed.

RESULTSFour ICMs from 7 blastocysts were cultured on MEFs. After culture, one cell line (cHES-1) was established and met the criteria for defining human pluripotent stem cells including a series of markers used to identify pluripotent stem cells, morphological similarity to primate embryonic stem cells and HES reported else where. Normal and stable karyotype maintained over 60 passages, and demonstrated ability to differentiate into a wide variety of cell types.

CONCLUSIONSHES cell lines can be established from gamete donors at a relatively highly efficient rate. The establishment will exert a widespread impact on biomedical research.

Blastocyst ; cytology ; Cell Differentiation ; Cell Line ; DNA-Binding Proteins ; analysis ; Female ; Fertilization in Vitro ; Humans ; Karyotyping ; Male ; Octamer Transcription Factor-3 ; Stem Cells ; cytology ; Tissue Donors ; Transcription Factors ; analysis

BACKGROUNDThe efficiency of traditional cryopreservation of human embryonic stem (ES) cells is low, and there have been few attempts to prove new cryopreservation methods effective. This study was designed to evaluate the efficiency of cryopreservation of human ES cells using vitrification method.

METHODSHuman ES cells clumped from an identical cell line were randomly allocated to be cryopreserved by vitrification or by slow freezing. The recovery rates, the growth and differentiation potential of thawed human ES cells were compared between these two groups. The pluripotency of human ES cells after thawing was identified.

RESULTSEighty-one point nine percent (59/72) of human ES cell clumps were recovered after vitrification, while only 22.8% (16/70) were recovered after slow freezing (P < 0.01). The colonies after vitrification manifested have not only faster growth but also a lower level of differentiation when compared to colonies subjected to the slow freezing protocol. However, the rates of growth and differentiation in undifferentiated colonies from both groups were identical to the rates in those of non-cryopreserved stem cells after a prolonged culture period. Passage 6 of vitrified human ES cells retained the properties of pluripotent cells, a normal karyotype and expressed the transcription factor OCT-4, stage specific expressed antigen-4 (SSEA-4) and SSEA-3. Teratoma growth of these cells demonstrated the ability to develop into all three germ layers.

CONCLUSIONSVitrification is effective in cryopreserving human ES cells. During a prolonged culture, human ES cells retain their pluripotency after cryopreservation.

Cell Differentiation ; Cell Survival ; Cryopreservation ; methods ; Embryo, Mammalian ; cytology ; Humans ; Osmotic Pressure ; Stem Cells ; cytology

OBJECTIVETo observe the effect of electroacupuncture (EA) on cytokines in the cardiac surgical patient and to evaluate the application of combined acupuncture anesthesia to cardiac surgery.

METHODSThirty patients with atrial septal defect were divided into 3 groups, general anesthesia group (A), acupuncture anesthesia group (B) and combined general anesthesia and EA group (C). Peripheral blood samples were collected before anesthesia, before cardiopulmonary bypass (CPB), 30 min after CPB and 24 h after operation to determine the levels of interferon-gamma (IFN-gamma), interleukin-2 (IL-2), IL-6 and IL-10.

RESULTSThe levels of IFN-gamma and IL-2 decreased in the 3 groups after CPB and further decreased 24 h after operation, and in the group C were higher than those in the group B. The levels of IL-6 and IL-10 significantly increased 24 h after operation in the 3 groups with no significant difference among the 3 groups.

CONCLUSIONThe general anesthesia combined with EA can not completely improve the decrease of IFN-gamma and IL-2 induced by CPB, indicating that the good response of the general anesthesia combined with EA to stress can partially improve the immunosupression induced by CPB. Acupuncture does not have significant effect on inflammatory cytokine reaction induced by cardiac surgery.

Acupuncture Analgesia ; Adult ; Cardiac Surgical Procedures ; Electroacupuncture ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Male ; Middle Aged

Objective To discuss the importance of nutritional intervention through monitoring the growth and the nutritional status of children with food allergy after comprehensive nutritional intervention.Methods A total of 158 children with food allergy who were under 2 years old were divided into an intervention group and an control group according to comprehensive nutritional intervention.The intervention group received diet guidance from a dietitian regu larly and took probiotics.If necessary,nutrients (calcium,iron and zinc) could be provided.The control group was given oral guidance by the pediatrician.After 6 months 1 follow-up,growth situation was assessed in terms of height and weight indexes,and nutritional intake condition was assessed in the light of serum biochemical indexes.Results The prevalence of malnutrition in the intervention group(17.6％) was significantly lower than that in the control group (37.0％),and there was a statistical sigmficance (x2 =22.03,P ＜ 0.05);the level of zinc [(88.5 ± 9.6) μmol/L]and iron [(18.6 ± 3.4) mmol/L] in the serum were significantly higher than those [zinc:(68.2 ± 8.5) μmol/L;iron:(12.8 ±2.7) mmol/L] in the control group,and there were statistical significance (zinc:t =12.264,P ＜ 0.01;iron:t =10.762,P ＜ 0.05);the daily intake of calcium[(596.1 ± 110.3) mg] and zinc [(10.7 ± 3.7) mg] in children (＞ 6-24 months) of the intervention group were significantly higher than those [calcium:(412.8 ± 89.3) mg and zinc:(7.3 ± 2.1) mg] in the control group (calcium:t =4.011,P ＜ 0.05,zinc:t =5.367,P ＜ 0.05),and the daily intake of protein [(42.8 ± 9.4) g] in the children (＞ 12-24 months) of the intervention group was significantly higher than that [(33.2 ± 8.2) g] in the control group,and there was a statistical significance (t =3.245,P ＜0.05).Conclusions Children with food allergy have risks of malnutrition,while comprehensive nutritional intervention can obviously decrease the prevalence of malnutrition and deficiency of iron,zinc and calcium,hence normative guidance of scientific feeding of children with food allergy is needed.

OBJECTIVETo developed a multiplex RT-PCR assay for simultaneous screening of type A, B and novel A (H1N1) influenza viruses.

METHODSTwo pairs of universal primers in were designed for amplifying the M gene and NS gene of type A and B influenza viruses, respectively. A pair of specific primers of HA gene was designed to detect novel A (H1N1) influenza virus. A one-step method was used to establish the multiplex RT-PCR system. A blinded experiment was carried out to validate the accuracy of this assay in comparison with the results of real-time fluorescence RT-PCR. The clinical practicability and efficacy of this assay was also evaluated.

RESULTSThe RT-PCR products were analyzed using agarose gel electrophoresis, which yielded distinct bands of the target fragments without non-specific reactions, suggesting the high efficiency and specificity of the multiplex RT-PCR. Blinded study of 50 samples demonstrated a concordance rate of 100%.

CONCLUSIONThis multiplex RT-PCR assay allows one-step simultaneous detection of type A, B and novel A (H1N1) influenza viruses rapidly and accurately, and provides a valuable low-cost screening technique for influenza epidemic monitoring and early diagnosis.

Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza B virus ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Time Factors ; Viral Matrix Proteins ; genetics ; Viral Nonstructural Proteins ; genetics

OBJECTIVETo investigate the polymorphic distribution of short tandem repeat (STR) sequences D21S1433, D21S1442, D21S1444, D21S2051 in Guangdong Han nationality in China.

METHODSUsing quantitative fluorescens PCR technology, the authors analyzed 200 unrelated samples to acknowledge the allele frequency, heterozygosity and other genetic information.

RESULTSD21S1433, D21S1442, D21S1444, D21S2051 were tested in 200 samples, which were tested to be statistical according to Hardy-Weinberg equilibrium (P> 0.05), 9, 10, 9 and 5 alleles were detected separately in each STRs. The heterozygosity of each STR was 0.818, 0.820, 0.770, and 0.261. The polymorphic information content > 0.7 in D21S1433, D21S1442, D21S1444, while D21S2051 owned only 0.247 polymorphic information.

CONCLUSIOND21S1433, D21S1442, D21S1444 are found to have high heterozygosity and polymorphic information content, and they could provide useful markers for genetic purposes, while D21S2051 is not informative in Guangdong Han nationality in China.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genetics, Population ; Heterozygote ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Pregnancy ; Tandem Repeat Sequences ; genetics

This study was purposed to investigate the role of post-thaw infused donor cells for predicting engraftment and hematopoietic reconstitution after unrelated cord blood transplantation (UCBT). The retrospective analysis was performed on clinical data of 97 children with malignant or non-malignant diseases received single unit UCBT from August 1999 to April 2010. The impact of pre-freezing and post-thaw cell dose of total nucleated cells (TNC), CD34(+) cells and colony-forming units-granulocyte/macrophage (CFU-GM) on engraftment and hematological recovery after UCBT was analyzed. Unrelated donors were from Guangzhou cord blood bank (GZCBB) entirely. The results indicated that the pre-freezing TNC (/kg) (mean ± SD: 7.65 × 10⁷ ± 4.26 × 10⁷; median: 6.34 × 10⁷), CD34(+)cells (/kg) (mean ± SD: 4.64 × 10(5) ± 4.47 × 10⁵; median: 3.03 × 10⁵) and CFU-GM (/kg) (mean ± SD: 0.79 × 10⁵ ± 1.09 × 10⁵; median: 0.57 × 10⁵) showed a good correlation with their post-thaw counterparts including TNC(/kg) (mean ± SD: 6.98 × 10⁷ ± 4.12 × 10⁷; median: 6.00 × 10⁷), CD34(+)cells (/kg)(Mean ± SD: 6.86 × 10⁵ ± 8.56 × 10⁵; Median: 4.17 × 10⁵), and CFU-GM (/kg) (mean ± SD: 0.52 × 10⁵ ± 0.52 × 10⁵; median: 0.39 × 10⁵) (r = 0.952, p < 0.001; r = 0.794, p < 0.001; r = 0.478, p < 0.001). Either the pre-freezing or post-thaw number of infused CFU-GM was significant higher in patients who achieved engraftment (n = 70) than those who suffered graft failure (n = 22) (p = 0.023 and 0.011, respectively), but no significant difference of TNC and CD34(+) cells dose (pre-freezing or post-thaw) were found between these two groups. Pre-freezing CFU-GM, TNC, CD34(+) cell dose negatively correlated with the time of neutrophil engraftment (r = -0.285, p = 0.018; r = -0.396, p = 0.002; r = -0.373, p = 0.002), as well as the post-thaw number of TNC and CD34(+) cells (r = -0.260, p = 0.031; r = -0.483, p < 0.001), whereas only pre-freezing CD34(+) cells showed a significant correlation with platelet engraftment time (r = -0.352, p = 0.013). It is concluded that the CFU-GM amount is useful for predicting engraftment of UCBT, while pre-freezing hematopoietic cell doses show superior correlation with the speed of engraftment and hematopoietic reconstitution than their post-thaw counterparts in pediatric recipients, suggesting that it is essential to perform hematopoietic potency assay on each cord blood unit prior to listing or release for administration.
Adolescent ; Antigens, CD34 ; blood ; Blood Banks ; Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Fetal Blood ; cytology ; Graft Survival ; Granulocyte-Macrophage Progenitor Cells ; Humans ; Infant ; Male ; Retrospective Studies ; Tissue Donors