BACKGROUND:Rabbit bone marrow mesenchymal stem cels as a kind of adult stem cels with strong proliferation and multilineage differentiation potential exhibit a tremendous application potential in tissue engineering and biological therapy.
OBJECTIVE:To in vitro culture, proliferate and identify rabbit bone marrow mesenchymal stem cels and to observe cel biological characteristics.
MEHTODS:Bone marrow of rabbits was extracted under sterile conditions to separate bone marrow mesenchymal stem cels using the whole bone marrow adherence method and Percol density gradient centrifugation method. Afterwards, the cels were purified and proliferated using differential adherence method. Morphology and growth pattern of cels were observed under microscope, and expression of cel surface antigen markers was detected by flow cytometry.
RESULTS AND CONCLUSION:Rabbit bone marrow mesenchymal stem cels presented with short adherent time and fast growth. After passage and purification, impurities cel counts were decreased. Primary cels presented with triangular, fusiform and spindly shapes. Passage 5 cels with single shape showed the typical polar swirling growth, and could not express CD34 and CD45, but expressed CD29 and CD44. These findings indicate that the cels cultured using the whole bone marrow adherence method and Percol density gradient centrifugation method possess stem cel characteristics in morphology, surface markers and multilineage differentiation, which have been identified as bone marrow mesenchymal stem cels by flow cytometry.

The resting cells of Citrobacter freundii 48003 3 expressing high tyrosine phenol lyase activity under the inducing of L tyrosine were used for L DOPA synthesis from catechol, pyruvate and ammonia In this paper, the effects of temperature, pH and substrate concentrations on the synthesis of L DOPA were studied At the optimal conditions of reaction, 9 5g/L of L DOPA was obtained in 12h

[ ABSTRACT] AIM: To observe the effect of circadian rhythms on wound healing of mouse corneal epithelium. METHODS:The C57BL/6 male mice were used in the study.A part of corneal epithelium (2 mm in diameter) was struck off by a golf-like knife to form a round wound area.The dynamics of epithelial healing in the wound area were ob-served under microscope with fluorescein staining.In addition, with related antibodies and DAPI, the dynamic changes of the neutrophils, platelets and dividing cells were also investigated.RESULTS:The healing rates in LL group (12 h light/12 h light) and DD group (12 h dark/12 h dark) were obviously slower than that in LD group (12 h light/12 h dark), mainly showing delayed re-epithelialization, decreased epithelial cells, increased diameter of blood vessel, and delayed re-cruitment of neutrophils and platelets, but more cell number.CONCLUSION:Disruption of circadian rhythms significant-ly inhibits the wound healing of corneal epithelium, mainly through delaying the inflammation and re-epithelialization, but aggravating the inflammatory responses.

Objective To explore the spiral the normal representations of permanent teeth and alveolar bone of multi-slice spiral(MSCT). Methods 30 adults with normal teeth were studied by multislice spiral CT scanner,window width and window level was 3000~5000 HU and 1200~1500 HU respectively.MSCT findings were analysed using multiplanar reformation,dental panorama and 3D reconstruction were done in all cases.Results The tooth were showed as multilaryer structures with different density at MSCT because of the mineral content of different part of tooth,from high to low density was enamel,circumpulpal dentin,outside dentin,cementum and pulp.The periodontal membrane was low density located beween the cementum and lamina dura and the thickness was homogeneous.The bone cortex of alveolar bone was successive,the bone trabecula appeared as wirelike or reticulate.Conclusion The structures of permanent teeth and alveolar bone can be displayed clearly by MSCT.Multiplanar reformation,dental panorama and 3D reconstruction are helpful to display the structures of teeth and alveolar bone as well as the relationship around structures.

Objective To discuss the importance of saving the life of patients and the severity assessment in early emergency for the trauma and hemorrhagic shock patients in the first-aid of pre-hospital and emergency department.Methods Retrospective anal-ysis of the 182 patients data.Results With early and aggressive treatment,177 cases of survival,5 cases of death,and the survival rate 97.3%.Conclusion The disease of traumatic shock patients is complex,high mortality.the early detection,taking timely and effective rescue measures are the key to increase the survival rate.

Objective To investigate the expression of miR-155 in rats exposed to PM2.5 and its signifi-cance.Methods Thirty-two 12-weeks-old healthy male SD rats were randomly divided into PM2.5-primary expo-sure group (P1),PM2.5-three times exposure group (P3), saline-control group and blank-control group (O), with 8 rats in each group. The pathological changes of lung tissue of each group were observed by HE staining and the expressions of miR-155 in lung tissue of each group were measured by RT-PCR. The expressions of TNF-α, IL-6 and IL-1β in serum of each group were measured by ELISA.One-way ANOVA and LSD-t test were used for statistical analysis,and Pearson′s correlation analysis was used to evaluate the relationship between the levels of miR-155 and TNF-α,IL-6 and IL-1β. Results The expression of inflammatory cytokines TNF-α,IL-6 and IL-1β in P1 and P3 increased when compared with that in saline control group and blank-control group, and the amount of expression increased with the increase of the number of exposure and dose;the difference between groups was statistically sig-nificant(P<0.01).At the same time,with the increase of the dose and number of PM2.5,the inflammatory damage of lung tissue was enhanced.The expression of miR-155 in lung tissue also increased with the increase of the dose and number of PM 2.5,and it had significantly positive correlation with serum inflammatory cytokines(P<0.01),and had statistical significance. Conclusions The lung tissue of rats exposed to PM2.5 can produce inflammatory injury, and the damage is enhanced with the increase of exposure times and dose.The expression of miR-155 is positively correlated with inflammatory injury in rats.Consequently,miR-155 may participate in PM2.5-induced inflammatory lung injury in rats.

Objective To explore the molecular pathological mechanism of gout, and to explore the mechanism of how interleukin (IL)-37 influencing PDZK1 protein in gout through nuclear factor κB (NF-κB) pathway. Methods HK-2 cells were stimulated with inflammatory signal IL-37. The expression of PDZK1 and its subcellular localization were detected by real-time fluorescence quantitative polymerase chain reaction (real-time PCR) at different concentrations of IL-37 (defined as group A), PDTC+IL-37 (defined as group B), Wortmannin+IL-37 (defined as group C), respectively.The changes of PDZK1 protein expression in HK-2 cells were detected by adding inhibitor PDTC (NF-κB pathway inhibitor) or Wortmannin (PI3K pathway inhibitor) and inflammatory signal stimulating protein imprinting method. The comparative t test was used for statistical analysis between groups A, B and A and C. Results The average levels of PDZK1 mRNA were as follows:(group A: 28.71 ±0.35, 28.57 ±0.31, 28.78 ±0.28, 28.63 ±0.29, 28.62 ±0.19; group B: 28.71 ±0.31, 28.83 ±0.27, 28.58±0.26, 28.73±0.36, 28.68±0.35;group C:28.81±0.32, 28.91±0.29, 28.72±0.24, 28.59±0.18, 28.58±0.22). There was no significant change in PDZK1 mRNA level in group A and B. The same IL-37 was found in group A and B. The values of T were 5.73, 4.72, 4.69, 5.86 and 6.79, respectively. The P values were all greater than 0.05, and there was no significant difference between the two groups. The values of T were 6.78, 7.13, 7.47, 6.38 and 5.98 in the same IL-37 concentration groups of A and C, respectively, and the P values were all greater than 0.05, with no significant difference. The levels of PDZK1 protein in the three groups by Western blot analysis were as follows: (group A: 0.200±0.082, 0.412±0.032, 0.723±0.063, 1.202±0.021, 1.222±0.023;group B: 0.124±0.064, 0.303±0.081, 0.325±0.062, 0.223±0.071, 0.343±0.052; group C: 0.017±0.022, 0.204± 0.015, 0.187±0.053, 0.302±0.051, 0.404±0.051), The levels of PDZK1 protein in group A treated with different concentrations of IL-37 followed by the concentration of IL-37. The level of PDZK1 protein in group B increased with the increase of IL-37 concentration, but the level of PDZK1 protein in group B was lower than that in the group treated with IL-37 only, and decreased when the concentration of IL-37 was 40 ng/ml.The level of PDZK1 protein in group C increased with the increase of IL-37 concentration, but the level of PDZK1 protein in group C was lower than that in the group treated with IL-37 only, and the same concentration of IL-37 in group A and B. The values of T were 1.83, 1.37, 1.64, 1.57 ,1.49, with P values greater than 0.05. There was no significant difference between the two groups. The values of T were 1.28, 1.37, 1.26, 1.42, 1.39 in the same concentration of IL-37 in group A and C, with P values greater than 0.05, with no significant difference.The results of immunofluorescence analysis showed that the trend of the three groups was basically consistent with that of Western-Blot. Conclusion The pathogenesis of gout induced by IL-37 through PDZK1 protein may not occur at the transcriptional level, but may occur at the level of protein translation.

Cystatin, a cysteine protease inhibitor found in many parasites, plays important roles in immune evasion. This study analyzed the molecular characteristics of a cystatin from Fasciola hepatica (FhCystatin) and expressed recombinant FhCystatin (rFhcystatin) to investigate the immune modulatory effects on lipopolysaccharide-induced proliferation, migration, cytokine secretion, nitric oxide (NO) production, and apoptosis in mouse macrophages. The FhCystatin gene encoded 116 amino acids and contained a conserved cystatin-like domain. rFhCystatin significantly inhibited the activity of cathepsin B. rFhCystatin bound to the surface of mouse RAW264.7 cells, significantly inhibited cell proliferation and promoted apoptosis. Moreover, rFhCystatin inhibited the expression of cellular nitric oxide, interleukin-6, and tumor necrosis factor-α, and promoted the expression of transforming growth factor-β and interleukin-10. These results showed that FhCystatin played an important role in regulating the activity of mouse macrophages. Our findings provide new insights into mechanisms underlying the immune evasion and contribute to the exploration of potential targets for the development of new drug to control F. hepatica infection.

OBJECTIVETo explore the common causative genes and mutation sites for hereditary non-syndromic deafness in Shanxi.

METHODSPeripheral blood samples were collected from regional schools for children with deafness. The samples were analyzed by matrix-assisted laser desorption ionization of flight mass spectrometry, and the results were verified by DNA sequencing.

RESULTSFor all samples, the 20 mutational sites of the 4 common causative genes were tested. As revealed, c.235delC of GJB2 gene has the highest mutational rate (13.67%). c.IVS7-2A>G of SLC26A (PDS) gene has a mutation rate of 17.67%, and c.1555A>G of mitochondrial 12S rRNA has a mutation rate of 2.00%. No mutations have been found with GJB3 gene. Sequencing analysis has suggested that the above results have a consistency rate of 99%.

CONCLUSIONAnalysis of mutations of the 4 common deafness-related genes can facilitate early diagnosis and treatment for the disease. Matrix-assisted laser desorption ionization time of flight mass spectrometry is a reliable method for such a task.

Adolescent ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; China ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; Deafness ; genetics ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation ; RNA, Ribosomal ; genetics ; Young Adult

Objective To analyze the drug consumption in medical service for Dominica and Dominican in Harmonious Mission -2018, and provide reference for future drug supply services in similar countries. Methods Data of the drugs used during the medical service of the two countries were extracted from the computer system, which were summarized and analyzed. Results Ranking first in drug consumption in medical service for Dominica and Dominican was external drugs. Chinese traditional medicines were very popular in both two countries. There were great similarities and unique characteristics in the drug consumption during medical service between the two countries, orthopedic drugs, digestive system drugs, ophthalmic drugs and dermatological drugs were frequently used drugs. Conclusion The drug consumption in medical service was closely associated with the national conditions. Therefore, in order to prepare medicines reasonably in future overseas medical service, disease distribution of the country which would be visited and drug consumption in the similar country which had been visited before should be concerned.