BACKGROUND: Is the inhibition of the hematopoietic stem progenitor cell (HSPC) proliferation and differentiation after human cytomegalovirus (HCMV) infection associated with abnormal expression of infected cell proliferated gene?OBJECTIVE: To observe the HOXB6-mRNA expression in the process of proliferation and differentiation of HCMV-infected HSPC into colony-forming unit granulocyte-macrophage (CFU-GM) and colony-forming unit erythroid (CFU-E).DESIGN: A controlled observation.SETTING: Laboratory for Molecular Biology, Affiliated Hospital of Luzhou Medical College, Lanzhou, Gansu Province, China.MATERIALS: All cord blood (CB) specimens were provided by the Obstetrics Department of Affiliated Hospital of Luzhon Medical College. They were collected from the umbilical vein of normal term neonates delivered spontaneously. All neonate mothers were healthy and HBS-Ag-negative. HCMV-IgM antibody revealed by routine ELUSA and HCMV-DNA checked by PCR were undetectable. Written informed consent for the laboratory measurements was obtained from each neonate mother, and the protocol was approved by the hospital's Ethics Committee. HCMV-AD169 strains were obtained from the Institute of Virology, Chinese Academy of Preventive Medicine. All-trans retinoic acid (ATRA, lot No. 20010126) was provided by Chongqing Huapont Pharm. Co., Ltd., China.METHODS: This study was performed at the Laboratory of Molecular Biology (state-level), Affiliated Hospital of Luzhou Medical College of Luzhou Medical College from April 2006 to April 2007. Cord blood mononuclear cells were separated for HSPC culture. According to different interventions, the study consisted of 4 groups. Control group: no HCMV virus solution was added and equal volume of culture medium was added instead. HCMV group: 105 PFU/mL HCMV-AD169 virus solution was added to the culture system. ATRA group: ATRA was added into the cultivation system at the final concentration of 60 μ mol/L. HCMV+ATRA group: ATRA was added into the HCMV group, and its final concentration was also 60 μ mol/L.MAIN OUTCOME MEASURES: In each group, cells were harvested on days 3,7 and 12. HOXB6 mRNA expression levels in CFU-GM and CFU-E were detected by real-time fluorescent-based quantification PCR.RESULTS: In the control group, both CFU-E and CFU-GM expressed HOXB6-mRNA. The HOXB6 mRNA expression was increased as a function of time. The HOXB6-mRNA expressed by CFU-E reached its peak level on day 12, while that expressed by CFU-GM reached its peak level on day 7. Compared to control group, the expression levels of CFU-E and CFU-GM HOXB6-mRNA genes in normal cord blood were significantly lower in the HCMV group (P＜0.05)and significantly higher in the ATRA group (P＜0.05) at each time point after HCMV infection. Furthermore, the expression levels were significantly higher in the ATRA+HCMV group than in the HCMV group at each time point(P＜0.05-0.01).CONCLUSION: HOXB6-mRNA expression is stable and lasting in the proliferation and differentiation of HSPC into CFU-GM and CFU-E. HCMV could down regulate HOXB6 gene expression, and ATRA could up regulate HOXB6 gene expression.

Objective To analyze the distribution of B-cell subtypes and IgG subclasses in pa-tients with non-small cell lung cancer ( NSCLC) and to investigate their potential functions in the progress of NSCLC.Methods Flow cytometry analysis was used to detect the distribution of immature B cells, memory B cells and mature B cells in 25 healthy subjects and 55 patients who were at different clinical stages of NSCLC.Enzyme-linked immune-sorbent assay ( ELASA) was performed to measure the concentrations of IgG1, IgG2, IgG3, IgG4 and total IgG, as well as the ratio of each IgG subclass in total IgG.Results The percentages of immature and memory B cells in CD19+B cells were positively correlated with the progress of NSCLC.The ratio of IgG4/IgG gradually increased along with the development of NSCLC, while that of the other three IgG subclasses showed no significant changes as compared with that of the control group.Conclu-sion The imbalanced distribution of B-cell subtypes and the up-regulated ratio of IgG4/IgG were closely as-sociated with the development of NSCLC.This study paved the way for further investigation on more effective immune therapy strategies targeting NSCLC.

Objective　To observe some biological features of bone marrow mesenchymal stem cells and explore the best conditions for isolatin g and culturing in vitro. Methods　Common cell culture techn ique, light and electron microscopy were used to study the effects of the growth , proliferation, morphology of the bone marrow mesenchymal stem cells in differe nt adherent time, concentration of serum and cell density. Results　The best culture condition in vitro for growth was 4-24 hours adherent time, 5%-10% fetal bovine serum, (4-8)×104/ml cell density. The cells were sp indle in shape and had a strong ability of proliferation. The time for cell duplication was 3 to 4 days. The cells showed the characteristics of stem cell s in electron microscope. Conclusion　The best condition for iso lation and culture of bone marrow mesemchymal stem cells was successfully establ ished and some biological features were obserred. It found a base for further in vestigation and using of mesenchymal stem cells.

Harpagoside (HAR) is believed to be a main compound in Scrophularia ningpoensis which possess a broad of biological activities.Human serum albumin (HSA) has important physiological roles in transportation, distribution and metabolism of many endogenous and exogenous substances in body.It is great significance in pharmacology to investigate the interaction mechanism of HAR and HSA.In this work, the interaction between HAR and HSA was investigated by fluorescence and ultraviolet absorption spectroscopy at different pH (pH=4.0, 7.4, and 9.0) and temperatures (297, 310 and 323 K).The experimental results showed that the HAR could cause the fluorescence quenching of HSA through a static quenching procedure, showing that the HAR regularly quenched the intrinsic fluorescence of HSA, and a decrease in the quenching constant was observed with an increase in temperature.Under different conditions, all the magnitude of binding constants (KA) was larger than 105 L/mol and the number of binding sites (n) in the binary system were approximate to 1.Base on the magnitude of enthalpy and entropy changes, the negative values of ΔG, ΔH and ΔS revealed that the binding of HAR with HSA was spontaneous and exothermic process, and the main interaction forces of the HAR with HAR were van der Waals forces and/or hydrogen bonding interaction.The binding distance (r) between the HAR and HSA was calculated to be about 4.2 nm based on the theory of F(o)rster′s nonradiation energy transfer, which indicated that the energy transfer from HSA to HAR occurred with high possibility.What was more, the synchronous florescence spectroscopy confirmed the conformational changes of HSA during the binding reaction.

Objective To clone platelet-derived growth factor A chain (PDGF-A) gene and insert PDGF-A gene into. Enhanced green fluorescent protein (EGFP) vector and then transformed into dermis-drived mesenchymal stem cells (DMSCs). Methods cDNA clones encoding human PDGF-A gene were isolated from a human hepatoma cell line mRNA by reverse transcription-polymerase chain reaction (RT-PCR). The PCR amplified fragment of PDGF-A gene was cloned into pMD18-T vector. The eukaryotic expression vector pEGFP-N1/PDGF-A was constructed by subcolone PDGF-A gene into pEGFP-N1 vector. PDGF-A gene was transfected into DMSCs with the help of Fugene 6 transfection reagent. Results Full cDNA sequence encoding human PDGF-A gene had been cloned, which sequence was consistent with the reported sequence in GenBank by sequence assaying. Conclusion cDNA sequence encoding human PDGF-A gene was successfully cloned into pEGFP-N1. The transient expression of PDGF-A gene in DMSCs has been realized.

ObjectiveTo study the effect of comprehensive treatment on lumbar interveretebral disc syndrome.Methods227 patients with lumbar interveretebral disc syndrome were randomized into two groups. Control group(127 patients) only accepted lumbar vertebra traction, while treatment group(100 patients) accepted comprehensive treatment, including lumbar vertebra traction, hot magnet, Maitland manipulation, and Mckeizie back muscles train. ResultsEffect of treatment group was obviously better than control group (P<0.05). Conclusions Comprehensive treatment on lumbar interveretebral disc syndrome can get better effect than simply traction.

Abstract: Objective To investigate the prevalence and pathogenic characteristics of Yersinia enterocolitica infection in children with diarrhea under 5 years of age in western Yunnan, and to provide a basis for the prevention and treatment of infectious diarrhea in children. Methods Feces were collected from under five-year-old children with diarrhea in the First Affiliated Hospital of Dali University from 2020 to 2021. Clinical information of the cases was also collected. Yersinia enterocolitica was isolated from the samples after cold enrichment on selective culture plates, and the pathogenic characteristics of Yersinia enterocolitica were analyzed by biological type and serotype and virulence gene detection. Results A total of 397 feces were collected. Seven strains of Yersinia enterocolitica were isolated in three samples, and the prevalence of Yersinia enterocolitica infection was 0.76% (3/397). Among the three positive samples, two Yersinia frederiksenii or Yersinia intermedia were isolated in specimen No. 212 , and five Yersinia enterocolitica were detected in specimens No. 24 and 226. Two Yersinia enterocolitica isolated from one sample were biological type 1A, and the virulence gene test results were ail-/ystA-/ ystB+ /yadA-/virF-, which were non-pathogenic Yersinia enterocolitica. Three Yersinia enterocolitica isolated from the other sample were biological type 3, serotype O∶3 (rfbc+), and virulence gene detection results were ail+/ystA+/ystB-/yadA+ /virF+, which were pathogenic Yersinia enterocolitica. While pathogenic Yersinia enterocolitica was detected from feces of children with diarrhea at 11 months of age with a infection rate of 0.50%(2/397). Conclusion Sporadic infection of pathogenic Yersinia enterocolitica was found in under five-year-old children in western Yunnan Province. It is necessary to strengthen the monitoring and research of Yersinia enterocolitica.

BackgroundDry eye is a multi-factorial-induced tear film and ocular surface disorder.Immunoinflammation plays a key role in the pathogenesis of dry eye.As inhibitor of the cyclo-oxygenase pathway,nonsteroidal anti-inflammatory drugs play an anti-inflammatory and anti-hypersensitivity role,and it can be a potential treatment for dry eyes.ObjectiveThis study was to investigate the effectiveness of nonsteroidal anti-inflammatory drugs (0.1％topical pranoprofen) on moderate to severe dry eyes and its mechanism.MethodsThis was a small sample of randomized controlled clinical trial.Thirty right eyes of 30 patients with moderate to severe dry eyes were included in the study according to the diagnosis criteria and randomized into two groups.The patients of the trial group received topical administration of 0.1％ pranoprofen plus 0.1％ sodium hyaluronate,and those of the control group received the topical 0.1％ sodium hyaluronate only.Ocular surface inflammation index scores (OSDI) and ocular surface fluorescine staining (OSS) scores were measured under the slit lamp,and tear film break-up time (BUT),Schirmer Ⅰ test values were evaluated.The expression of human leucocyte antigen-DR (HLA-DR) and CD11b in conjunctiva epithelial cells were detected by impression cytology and flow cytometry (FCM).All the indexes were compared between the two groups before and after treatment.Informed consent was obtained from all patients.ResultsThere were no significant differences in terms of age and gender and their baseline values between the trial group and control group (t=0.412,P=0.684;x2=0.240,P=0.624),and so were all the indexes (P＞0.05).Compared with the control group,the OSDI,OSS scores and cells positive for HLA-DR were lowered but the BUT was delayed in the trial group on day 15 ( t=2.43,P=0.03;t=2.83,P=0.01;t=3.29,P=0.00;t=3.23,P=0.00 ).No significant differences were found in the Schirmer Ⅰ test value and CD11b expression between these two groups (t=0.17,P=0.87;t=0.28,P=0.79).The OSDI,OSS scores and BUT were significantly improved,and the number of cells positive for HLA-DR were reduced 15 days after administration of drugs in comparison with before treatment in the trial group ( t =12.30,10.70,6.10,7.92,P =0.00 ).However,there were no comparable alteration seen in these indexes before and after the usage of drugs in the control group ( P＞0.05).Positive correlations were found in HLADR expression with OSDI and OSS ( r =0.601,P =0.018 ; r =0.586,P =0.022 ) and a negative correlation in HLADR expression with BUT (r=-0.697,P=0.004) on day 15 in the trial group.ConclusionsTopical usage of 0.1％ pranoprofen is beneficial for remitting the ocular signs and symptoms in moderate to severe dry eyes.This study illustrates that topical usage of 0.1％ pranoprofen can down-regulate the expression of inflammatory markers in conjunctival epithelial cells.

No abstract available.
Colitis, Ulcerative* ; Ulcer*

Objective To observe whether the transplanted dermal multipotent stem cells(dMSCs)transfected by adenovirus vector of CXCR4(Adv-CXCR4)can distribute more frequently to the wound of rats with combined wound and irradiation injury.Methods dMSCs transfected by Adv-CXCR4(group A),or transfected by adenovirus vector of green fluorescent protein(group B),and non-transfected dMSCs were labeled with 3H-TdR and then transplanted into combine-injured rats.The amount of dMSCs in wound were determined by liquid scintillation,and wounds healing process was observed by measuring the remaining wound area.Results From the 5th day after transplantation,the amount of dMSCs in the wound of group A accounted for 1.95%-3.85% of the total transplanted dMSCs,significantly greater than those in group B and group C,which accounted for 1.07%-1.86% of the total transplanted dMSCs.The remaining wound area in group A was smaller than those in group B and group C from day 12 after injury,and the healing time of group A was 1.5 day ahead than group B and group C.Conclusions dMSCs transfected by Adv-CXCR4 distributes more frequently to the wound of combine-injured rats and could accelerate wound healing.