Objective To investigate the development and dynamic changes of host immune response in DBA/2 mice infected with Plasmodium yoelii 17XL. Methods Female DBA/2 mice were infected by intraperitoneal ( i. p. ) injection of 106 P. yoelii 17XL parasitized erythrocytes ( PRBC). Levels of IL-12, IFN-γ, IL-4, IL-10 and P. yoelii 17XL-specific antibody in sera were measured by ELISA. Concentrations of NO in cell supernatants were measured by the Griess reaction. Parasitemia,percentage of mononuclear-macrophages of individual mice were monitored daily, and phagocytosis of mononuclear macrophages was also observed. Results Primary parasitemia in vein blood was developed on day 3 postinfection, which peaked with a level of 46. 9% on day 9. Most mice cleared the infection and survived by day 20 postinfection. From day 6 to day 16, the phagocytosis of PRBC by rodent macrophages was observed on the blood smear. Infected mice had a continuously increased level of IL-12 in serum from day 1 postinfection. Accordingly, high level of IFN-γ was also detected in sera from day 1 postinfection,which peaked on day 6. Infected mice produced higher level of IL-4 and IL-10 in serum on day 6 postinfection, which peaked on day 9 and day 15 postinfection respectively. In addition, splenocytes from infected mice produced significantly higher level of NO on day 6 and 20 postinfection. Level of P. yoelii 17XL-specific IgG was determined in the sera of infected mice with a steadily increased trend after infection, which peaked on day 70 postinfection. Conclusions Effective polarizing of Thl cells is significant in inhibition of parasitemia and eventual clearance of the Plasmodium parasites. Activated mononuclear-macrophages play a key role in inhibiting parasitemia in the early phase of infection with P. yoelii 17XL.

Objective To investigate the location,fine structure of melanocytes in human fetal scalp hair follicles.Methods The scalp with hair follicles was obtained from a dead fetus of 6 months of age,and divided into two parts.One part was embedded in paraffin,tissue sections were prepared with a width of 7 μm and stained with NKI/beteb,monoclonal antibodies to HMB-45,tyrosinase and tyrosinase-related protein 1(TRP1),respectively.The other part with hair follicles was treated with collagenase type Ⅱ 0.1 g/L and trypsin,then,cell suspension was collected and cultured.After 14-day culture,follicle melanocyte cells (FMC)were separated from keratinocytes by differential trypsinization,and fibroblasts were removed with geneticin.Following three times of pure passage,FMC were seeded and fixed on mica for scanning electron microscopy(SEM)and atomic force microscope(AFM)scanning.Results Histopathological examination showed that NKI/beteb positive cells located at the outer root sheath of human hair follicles,and these cells stained negatively for HMB-45,tyrosinase and TRP1 antibodies.However,in the hair bulb,lots of cells expressed HMB-45,tyrosinase and TRP1 antigens.After fibroblasts and keratinocytes were removed,two kinds of melanocytes remained in the culture:one was small in number and showed abundant melanin,which was lost after subsequent passage;the othgr was large in number and had no melanin initially,but proliferated very rapidly.After three passages,almost all the melanocytes were positive for NKI/beteb.As SEM and AFM showed,most cultured melanocytes appeared fusiform with two(rarely three)dendrites,and the cell body was round or oval with a few melanosomes scattered in but no clear secondary branches on the dendrites.Conclusions The melanocytes in outer root sheath of hair follicles from the fetal scalp are presumed as melanocyte stem cells or their progenies.In vitro,these cells proliferate very rapidly during early phases,but the morphology and function of them still remain immature,which is unfavorable for melanosome transport.

Objective To evaluate the stability of dipstick dye immunoassay (DDIA) kit forschisitosomiasis diagnosis. Methods By means of detection of the sera from infected people withSchistosoma japonicum and healthy people, the stability of the DDIA kit, which stored at 37℃,room temperature or 4 ℃ respectively, was evaluated depending on the detective results ofsensitivity, specificity, detectable minimum and coefficient variation ( CV). Results Thesensitivity, specificity, detectable minimum and coefficient variation of the DDIA kit were invariableafter the kits stored at 37 ℃ for 180 days, and at room temperature or 4 ℃ for 360 days.Conclusion The DDIA kit is stable while it stores at 37℃ for 180 days, and at room temperatureor 4℃ for 360 days at least.

Objective To optimize the extraction process of Albizia julibrissin in Baijin Capsule. Methods Using the extraction rate of quercitrin and total flavonoids as indexes, the orthogonal design was used to investigate effects of solvent volume, extraction time and extraction frequency on extraction results. Results The optimal extracting condition was as follows:extracted 2 times with 12 fold 70%alcohol, 2 h for each time. Conclusion The optimized process condition was simple, stable and feasible. It provides the basis for the production of Baijin Capsule.

BACKGROUND:Some studies have shown that more copy number variations are present in early passage human induced pluripotent stem cells than later passage human human induced pluripotent stem cells, their parental somatic fibroblasts or human embryonic stem cells. OBJECTIVE:To investigate whether the reprogramming process itself compromises genomic stability and further explore the efficiency of induced pluripotent stem cellestablishment. METHODS:Using high-resolution Affymetrix CytoScan HD array, we compared copy number variations and loss of heterozygosity in early passage induced pluripotent stem cells with their fibroblast cellorigins from genetic epilepsy patients. RESULTS AND CONCLUSION:Compared with somatic fibroblasts from genetic epilepsy patient, there was no difference in the loss of heterozygosity between the two types of cells, but more copy number variations were present in early passage human induced pluripotent stem cells which were characterized as microduplication and involved oncogenic genes. Results demonstrate the dynamic nature of genomic abnormalities during reprogramming process and the necessity of frequent monitoring human induced pluripotent stem cells to assure their genomic stability and clinical safety.

AIM:To study the coating technique of pH-time lap colon-specific matrine delivery mini-pill consisted of the time lag release coating (inner layer) and enteric coating (out layer). METHODS:To filter the coating composition based on the index of dissolution of matrine and oxymatrine in vitro and the appearance rating of miui-pill. RESULTS:4 The coating composition of inner layer was the alcoholic solution,consisted of 2% EC,0.4% DEP and 2% talc powder. Then the coating composition of out layer was the alcoholic solution consisted of 5% Eudragit S100,3% talc powder and 0.5% TEC. The dissolution tests in vitro indicated that matrine and oxymatrine were not dissolved in the simulated gastric juice in 2 h. The accumulative amount of matrine and oxymatrine were less than 15% in the simulated intestinal fluid in 4 h. The amount of matrine and oxymatrine were 80.7% and 83.5% in the simulated colon juice in 2 h. CONCLUSION:The mini-pill could achieve the goal of delivering in the specific colon.

To construct retroviral vector carrying human interleukin-3 c0mplementary DNA(HuIL-3 cDNA) under control of human a-fetoprotein gene enhancer core sequence and human SV4O pro-moter. Methods and Results: HuIL-3 cDNA was inserted into polylinker site of retroviral vector pMNSMto construct retr0viral vector pMNS-IL-3, in which the transcription of HuIL-3 cDNA was drived by SV40early region promoter. Human Q-fetoprotein gene enhancer core sequence was released from plasmidpGEM. 7Zf-AFPe and inserted into the polylinker site of pMNSM. Then human interleukin-3 cDNA wasinserted int0 p0lylinker site to construct retroviral vector pMNSA-IL-3, in which HuIL-3 cDNA transcrip-tion was drived by SV40 early region promoter and enhanced by human a-fetoprotein enhancer core se-quence- Conclusion: The vectors are of significance for hepatoma-specific gene therapy.

ive] To explore a new method of evaluating the healing of fractures. [Methods] Self-con trolled trial was applied. Bone mineral density ( BMD) of fracture end (region 3, R 3 ) and its neighboring proximal and distal regions ( R 2 and R 1 respectively) in 126 cases of fracture of lower end of radius was deter mined by dual-energy X-ray absorptionmetry. [Results] BMD of R3 elevated as compared with that of R 2 and R1 (P

BACKGROUND:The induced concentration for osteoblasts is often introduced as reference to induce odontoblast differentiation. However, there are no reports on other concentrations. OBJECTIVE: To observe the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein during low-dose β-glycerophosphate-induced differentiation of dental pulp stem cels into odontoblasts. METHODS:Human dental pulp stem cels were isolated and cultured, and then induced by different concentrations of inducing solution to differentiate into adipocytes and osteoblasts, which could verify the multi-directional differentiation ability of human dental pulp stem cels. Under 5 mmol/L β-glycerophosphate, dental pulp stem cels differentiated into odontoblasts. At 7, 14, 21, 28 days of culture, RNA samples were extracted from dental pulp stem cels in each group, and reverse-transcription PCR was used to detect the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein. Mineralized nodules were detected by alizarin red S staining to show the successfuly osteogenesis induction. RESULTS AND CONCLUSION: Human dental pulp stem cels could be induced to adipocytes and osteoblasts. The results of reverse-transcription PCR showed that the dental pulp stem cels under 5 mmol/L β-glycerophosphate could increase the expression of dentin matrix protein-1 and dentin sialoprotein, but downregulate the expression of matrix extracelular phosphoglycoprotein at 7, 14, 21 days. At 28 days of culture, dental pulp stem cels were al successfuly mineralized detected by alizarin red S. There were some red mineralized nodules. These findings indicate that the 5 mmol/L β-glycerophosphate can induce the differentiation of dental pulp stem cels into odontoblasts successfuly, up-regulate the mRNA expression of dentin sialoprotein and dentin matrix protein-1, and meanwhile down-regulate the mRNA expression of matrix extracelular phosphoglycoprotein.

Objective:To observe the effects of warming needle therapy combined with the modified Hua Tan Tong Luo Tang(decoction for dissipating phlegm and unblocking collaterals)on post-stroke cognitive impairment due to obstruction of wind-phlegm. Methods:A total of 78 patients who met the inclusion criteria were randomly assigned to an observation group and a control group,with 39 patients in each group.Both groups received conventional treatment.The control group was additionally treated with the modified Hua Tan Tong Luo Tang,while the observation group received warming needle therapy in addition to the treatment given to the control group.Scores on the mini-mental state examination(MMSE),Montreal cognitive assessment(MoCA),and activities of daily living(ADL)were assessed before and after treatment.Adverse reactions and clinical efficacy were analyzed and compared between the two groups after treatment. Results:The total effective rate was 82.1%in the observation group and 61.5%in the control group,showing statistical significance between the two groups(P<0.05).After treatment,the MMSE,MoCA,and ADL scores of both groups were significantly higher than those before treatment within each group(P<0.05).Notably,the scores in the observation group were higher than those in the control group(P<0.05).No adverse reactions occurred in either group during the treatment period. Conclusion:Based on the conventional treatment,compared to modified Hua Tan Tong Luo Tang alone,warming needle therapy combined with the modified Hua Tan Tong Luo Tang showed superior clinical efficacy in treating post-stroke cognitive impairment due to obstruction of wind-phlegm,and this combined therapy also showed high safety.