Objective To evaluate the effect of C-type natriuretic peptide (CNP) on paraquat-induced pulmonary fibrosis in mice.Methods Fifty-four male Kunming mice,aged 8-10 weeks,weighing 30-35 g,were randomly divided into 3 groups (n=18 each) using a random number table:control group (C group),pulmonary fibrosis group (PF group) and CNP group.Paraquat 10 mg/kg (in 0.1 ml of normal saline) was injected intraperitoneally once every 3 days for 5 times in total in PF and CNP groups,and in addition CNP 3 μg/kg (in 0.1 ml of normal saline) was simultaneously injected via the tail vein once every 2 days for 14 times in total in group CNP.The equal volume of normal saline was given instead of paraquat in group C.On days 1,8 and 15 after the end of administation of paraquat,6 mice were sacrificed,and lungs were removed for determination of wet to dry weight ratio (W/D ratio),hydroxyproline (HYP) content (using alkaline hydrolysis),and transforming growth factor-beta 1 (TGF-β1) content (using enzymelinked immunosorbent assay) in lung tissues.Results Compared with group C,the W/D ratio and contents of HYP and TGF-β1 in lung tissues were significantly increased at each time point after the end of administation of paraquat (P＜0.05),and the pulmonary fibrosis was obvious in group PF.Compared with group PF,the W/D ratio and contents of HYP and TGF-β1 in lung tissues were significantly decreased at each time point after the end of administation of paraquat (P＜0.05),and the pulmonary fibrosis was significantly attenuated in group CNP.Conclusion CNP can reduce paraquat-induced pulmonary fibrosis in mice.

Objective To observe the repairing effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) and goserelin on chemotherapy-induced ovarian injury, and the distribution and growth of hUC-MSCs transplanted in rat chemotherapy-induced ovarian injury. Methods A total of 120 SD rats were randomized into group A-E:A normal group, B NS control group, C goserelin group, D hUC-MSCs group and E hUC-MSCs+goserelin group. The rat premature ovarian failure (POF) model was established by given a loading dose of cyclophosphamide (CTX, 50 mg/kg) followed by daily intraperitoneal injection of CTX (8 mg/kg) for consecutive 14-day. The hUC-MSCs were injected through caudal vein, and goserelin was given by subcutaneous injection 4 days before POF model established. The serum level of estrogen was detected and numbers of follicles were counted. After GFP was transfected by lentivirus, the distribution and growth of stem cells transplanted in rats were observed by animal in vivo imaging system. Results At day 46, the serum level of estrogen showed no significant difference between group A and group E (P > 0.05). There were no significant differences in the counted follicles between group A and group E (P>0.05). After tail vein injection of the transfected cells, GFP positive cells were found in injury ovarian. Conclusion There is a repairing effect of hUC-MSCs and goserelin on ovarian injury.

Objective To observe the effect of hydrogen on ultraviolet B (UVB)-induced oxidative damage to skin fibroblasts.Methods Primary human skin fibroblasts from foreskin tissues were divided into five groups:normal control group receiving no treatment,hydrogen control group treated with hydrogen-rich saline,UVB group receiving irradiation only,post-treatment group irradiated with UVB followed by hydrogen-rich saline treatment,and pre-treatment group treated with hydrogen-rich saline followed by UVB irradiation.The dose of UVB was 30,60 and 90 mJ/cm2 in the cell proliferation assay and 90 mJ/cm2 in the other experiments.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of fibroblasts,a chemiluminescence method to estimate the activity of superoxide dismutase (SOD) and catalase as well as to determine the level of malondialdehyde in the culture supernatant of fibroblasts,enzyme linked immunosorbent assay (ELISA) to determine the supernatant level of 8-isoprostane-prostaglandin F2α (8-iso-PGF2α),Western blot to detect the expression of heme oxygenase-1 (HO-1) in fibroblasts.One-factor analysis of variance was conducted to assess differences in these parameters among these groups.Results UVB irradiation decreased the proliferative activity (absorbence value at 490 nm) of fibroblasts in a dose-dependent manner.Both the pre-treatment group and post-treatment group showed a statistical increase in proliferative activity of cells compared with the corresponding UVB control groups (all P ＜ 0.05).The activity of SOD and catalase as well as the protein expression of HO-1 were significantly higher (all P ＜ 0.05),whereas the supernatant levels of malondialdehyde and 8-iso-PGF2α were statistically lower (both P ＜ 0.05) in the pre-treatment group and post-treatment group than in the UVB control group.Conclusion Hydrogen may mitigate UVB-induced oxidative damage to skin fibroblasts.

Implant surface modified coating can improve its osteoinductivity, about which simple calcium phosphate coating has been extensively studied. But it has slow osteointegration speed and poor antibacterial property, while other metal ions added, such as nano zinc ion, can compensate for these deficiencies. This paper describes the incorporation form, the effect on physical and chemical properties of the material and the antibacterial property of nano zinc, and summarizes the material's biological property given by calcium ion, zinc ion and inorganic phosphate (Pi), mainly focusing on the influence of these three inorganic ions on osteoblast proliferation, differentiation, protein synthesis and matrix mineralization in order to present the positive function of zinc doped calcium phosphate in the field of bone formation.
Biocompatible Materials ; Calcium ; Calcium Phosphates ; chemistry ; Cell Differentiation ; Cell Proliferation ; Humans ; Ions ; Metal Nanoparticles ; chemistry ; Osteoblasts ; cytology ; Osteogenesis ; Phosphates ; chemistry ; Zinc ; chemistry

Objective:To evaluate any effect of transcranial pulsed current stimulation (tPCS) on the motor functioning of rats modelling stroke using the Catwalk gait analysis system.Methods:A stroke model was induced in 24 rats using middle cerebral artery embolization. They were then randomly divided into a sham operation group, a model group and a tPCS group, each of 8. Neurological deficit scores were assigned 1 day after the modeling. Beginning two days after the modeling the tPCS group was given 20 minutes of tPCS daily with an intensity of 0.2mA at 10Hz for 7 days. Gait data were collected using the Catwalk gait system 1 day before, as well as 1 and 9 days after the modeling.Results:Nine days after the modeling the average Bederson neuroethology score of the tPCS group was significantly lower than one day after the modelling and significantly lower than the model group′s average. One day after the modelling significant differences were observed in the model and tPCS groups in the average contact area of the affected limb′s paw prints, limb swing speed, stride length, limb speed, swing time, average running speed and standing time compared with before the operation. After nine days the average standing time on the affected fore and hind limbs, as well as the paw contact areas were significantly better in the tPCS group than in the model group.Conclusion:tPCS can promote improvements in gait after ischemia and reperfusion, at least in rats.

Objective To investigate the regulation of T cells in the process of liver regeneration using a model of mice after 70% liver resection.Methods We performed 70% hepatectomy in T-cell-deficient mice and control mice.The liver mass and body mass ratio, BrdU infiltration level, proliferating cell nuclear antigen (PCNA),expression of M phase marker protein p-HDAC3, and serum transaminase levels were measured.Results The recovery of liver mass and body mass ratio of thymus-deficient mice occurred significantly later than that of control mice.The peak time of BrdU infiltration levels and the expression of PCNA and p-HDAC3 in T-cell-deficient mice were later than in control mice, but the degree of liver injury was lower.Conclusion T cells are involved in the regulation of liver regeneration, and the absence of T cells delays the process of liver regeneration.

Objective To determine the effect of NEL-like type 1 gene (NELL-1) transfection in vivo in the repair of traumatic femoral head necrosis.Methods Twenty-four SD rats were randomly divided into three groups (8 rats per group) according to the lottery method,ie,sham group (served as normal control),NELL-1 treatment group (injected NELL-1 gene by recombinant adenovirus vectors around the hip one week after osteonecrosis model induced surgically) and placebo group (given an equal volume of saline solution at the same time after the induction of osteonecrosis).Femurs were taken from the animals 5 weeks after surgery.Gross observation was performed for morphology changes,X-ray assessment for femoral head height and length ratio (H/L),Micro-CT measure for bone parameters of femoral head including total volume (TV),bone volume (BV),total mineralized content (TMC),trabecular thickness (Tb.Th) and trabecular space (Tb.SP),and histological study for osteocytes,osteoblasts and osteoclasts.Results Preserved femoral head shape was noted in NELL-1 treatment group compared to the obvious flattening of the femoral head in placebo group.No heterotopic osteogenesis was observed in any group.Femoral head H/L ratio for 0.753 2 ± 0.040 2 in NELL-1 treatment group was higher than 0.598 4 ± 0.037 0 in placebo group (P ＜ 0.05),but lower than 0.920 2 ± 0.037 0 in sham group (P＜0.05).TV,BV,TMC and BMD between NELL-1 treatment and sham groups did not differ significantly (P ＞ 0.05),but all were increased compared to placebo group (P ＜ 0.05).There was no significant differences in Tb.Th and Tb.SP among three groups (P ＞ 0.05).Most osteocytes were alive in NELL-1 treatment group.More active osteoblasts and osteoclasts were noted in NELL-1 treatment group than those in placebo group.Conclusion NELL-1 gene transfection can preserve femoral head shape and bone content,promote osteoblast activity and neovascularization and hence is an effective treatment for rat traumatic osteonecrosis.However,the activity of osteoclasts is stimulated simultaneously.

Objective To study the effect of inflammatory markers on the level of reactive oxygen species (ROS) and mitochondrial DNA (mtDNA) copy numbers in granulosa cells of patients without polycystic ovary syndrome (PCOS). Methods Fifty patients without PCOS treated with in vitro fertilization and embryo transfer (IVF-ET) were selected in this study. The granulosa cells were extracted and cultured in vitro. Cells were randomly divided into treatment group and control group. The 5 nmol/L interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF)-αwere given to treatment group, and same amount of inflammatory diluted solution was added to control group. The levels of ROS and copy numbers of mtDNA were compared between two groups. Results The ROS levels and mtDNA copy number of granulosa cells were significantly higher in IL-1, IL-6 and TNF-αtreatment groups than those of control group (P<0.05). Conclusion Inflammatory markers of IL-1, IL-6 and TNF-αincrease the level of ROS and damage mtDNA in granulosa cells.

Objectives To study the performance of different microbial automated inoculation systems and to evaluate the performance of the Probact microbial automated inoculation and incubation system ( Probact system) and its applications in clinical microbiology laboratory.Methods A total of 160 clinical specimens, including respiratory secretions ( n=61 ) , urine ( n=49 ) , and feces ( n=50 ) , that were submitted to the Clinical Microbiology Laboratory in Peking Union Medical College Hospital of Chinese Academy of Medical Sciences from February 2015 to April 2015 were evaluated.These specimens were processed with conventional manual method, the Probact automated inoculation system, and PREVI Isola Inoculator.The quantity of bacterial species recovery, number of effectively isolated colonies, total number of colonies recovery per plate, and time of processing the 160 specimens by the three methods were evaluated. Wilcoxon signed-rank test and Kruskal-Wallis rank sum test were used for statistical analysis.Results The Probact system had significantly higher quantity of bacterial species recovery (respiratory specimens 3.41 ±1.40, urine 1.92 ±0.86, and feces 1.16 ±0.79) than those by the Isola Inoculator (respiratory specimens 3.75 ±1.29, urine 2.24 ±0.97, and feces 1.92 ±0.72), (P=0.006, 0.011, <0.001).Compared to the manual method, Probact performed less quantity of bacterial species recovery for respiratory specimens(3.85 ±1.38), but higher in feces(0.80 ±0.81)( P<0.001).There is no significant differences for urine ( 1.84 ±1.23 ) ( P=0.266 ) .As for number of isolated colony, the Probact system ( respiratory specimens 12.16 ±7.72, urine 2.71 ±4.24, and feces 5.40 ±5.04 ) had significant smaller numbers than that of Isola Inoculator (respiratory specimens 16.56 ±5.76, urine 4.35 ± 4.89, and feces 8.40 ±3.70) (P<0.001,0.007,0.003).However, both system had larger numbers of isolated colonies than those by the manual method (respiratory specimens 11.30 ±8.42, urine 2.67 ±4.34, and feces 1.90 ±3.90) and the difference was significant for fecal specimens(P<0.001).Regarding the total number of colonies recovery, larger number was found by Isola Inoculator than that by the Probact system for fecal specimens, however, there were no significant differences for respiratory or urine specimens (P=0.524,0.738).Compared with manual method, the Probact system had significantly more numbers of colonies recovery for respiratory and fecal specimens ( P<0.001 ) . The total time for processing 160 specimens was shortest for manual method (281 min), followed by Probact system (419 min) and Isola Inoculator (495 min) .Conclusions The performance of the Probact system is better than the manual method but no superior to the Isola Inoculator.The Probact system can meet the clinical need in terms of full automation and standardization of specimen inoculation and prevention of bias of processing by laboratory staffs using manual method.

Objective:To express the glycoprotein D of herpes simplex virus type 2 (gD2) in the insect cells,and to determine its immunogenicity.Methods:HSV-2 genome was used as the template for amplification of gD2 extracellular domain fragment gene by PCR.The PCR product was inserted into the vector Bacmind,and the constructed recombinant plasmid gD2-Bacmind was transfected into the sf9 cells to package the recombinant baculovirus.The Sf9 cells were infected by recombinant baculovirus seed derived from the forth passage(P4),the titer of P4 recombinant baculovirus was detected by a plaque assay and the expression of recombinant protein gD2 was determined by Western blotting method.The supernatant of infected cells was collected and purified by Ni-NTA affinity chromatography to obtain the target protein gD2,the purified gD2 protein was used to immunize the BALB/c mice in 0, 2, 4 weeks (gD2 group),and PBS was used as negative control(PBS group);the titers of gD2 specific IgG in serum were detected by ELISA assay.Results: The PCR analysis and sequencing results proved that gD2-Bacmind was constructed correctly.The titer of recombinant baculovirus was 2.0×109 pfu·mL-1,the purified gD2 was about 37 000 with expectation,the percentage of gD2 in total protein was 90%.The average value of Log10 of titer of gD2 specific IgG in serum detected by ELISA assay in gD2 group at the sixth week was 4.34,and there was significant difference compared with PBS group(P<0.01).Conclusion: The gD2 expressed by insect-baculovirus expression vector system has the immunogenicity and can be selected as candidate protein for HSV-2 vaccine.