Objective To investigate the effect of matrix metalloproteinases(MMPs) in ventilator-induced lung injury inflammatory reaction of newborn rabbits.Methods Seventy-five newborn rabbits aged 1-5 days were randomly arranged to 3 groups:control group(n=3),other 72 rabbits designed by 2?2?3 factorial were divided into high concentration oxygen(100%)and low concentration oxygen(45%) groups,and each group was subdivided into 2 groups:High peak inspiratory pressure(PIP)group and low PIP group.All rabbits were done by mechanical ventilation and killed to obtain lung tissue at 1,3,6 hours after trailing respectively.The concentration of MMP-9 and MMP-2 in lung tissue bomogenate were detected by ELISA,at the same time the ratio of wet to dry was detected and pathological section was analyzed.Results 1.The concentration of MMP-2 in high concentration oxygen group,high PIP group and low PIP group increased compared with that in normal group,and there were significant differences.2.The mean concentration of MMP-9 was lower in high concentration oxygen group than that in normal control group,and there was significant difference between 2 groups.The mean concentration of MMP-9 in low concentration oxygen group was higher than that in normal group,and there was no significant difference between 2 groups.There was no significant difference in effect to mean concentration of MMP-9 by different PIP.3.There were positive correlation between MMP-2 and MMP-9,MMP-2 and the ratio of wet to dry in lung.Conclusions Within 6 hours in newborn rabbits by mechanical ventilation,ventilation with high concentration oxygen up-regulate MMP-2,but down-regulated MMP-9.The lungs stretch by mechanical ventilation increase MMP-9,but decrease MMP-2.Mechanical ventilation can affect the synthesis of MMPs,and MMPs plays an important role in ventilator-induced lung injury of newborn rabbits.

OBJECTIVETo observe the effect of tensile stress on human heel skin fibroblast proliferation in vitro, providing a theoretical basis for preventing the wound edge skin necrosis and nonunion after calcaneal fracture surgery.

METHODSFibroblast cells were taken from lateral heel skin of a 40 year-old-man, then cultured and subcultured in vitro. After that, they were divided into three groups: 0 hours group, 6 hours group and 24 hours group and were tested by tensile stress testing. The levels of TGF-β1 and IL-6 in nutrient fluid were measured. Transmission electron microscope and light microscope was applied for observe mitochondria and nucleus.

RESULTSUnder 10% of the tensile stress, mitochondria decreased, the levels of TGF-β1 and IL-6 in nutrient fluid were decreased and cell proliferation was inhibited gradually with time increasing.

CONCLUSIONThe human lateral heel skin in a long-time tensile stress state is an important cause of wound edge skin necrosis and nonunion after calcaneus fracture surgery.

Adult ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; chemistry ; cytology ; Heel ; physiology ; Humans ; In Vitro Techniques ; Interleukin-6 ; metabolism ; Male ; Skin ; chemistry ; cytology ; metabolism ; Tensile Strength ; Transforming Growth Factor beta1 ; metabolism

The system of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated nuclease (Cas) 9 is an effective tool for revising the genome with great accuracy,and boost the advances in life science.By employing this system,we discover the regulation role of key gene during retina development and correct the abnormal mutation of these genes.In this paper,we summarize CRISPR-based technologies that enable mammalian genome editing and their various applications.And CRISPR/Cas9 may be a promising tool to disclosure the mechanism of retinal diseases so as to develop novel treatment for patients with retinitis pigmentosa.

OBJECTIVETo detect angiotensin II (ANG II) in the maternal blood, umbilical cord blood, and at the maternal-fetal interface in women with pregnancy-induced hypertension (PIH) and explore the etiology of PIH and pathophysiologic mechanism of intrauterine growth restriction (IUGR).

METHODSEnzyme-linked immunosorbent assay (ELISA) was used to detect the concentration of ANG II in the maternal blood, cord blood, and maternal and fetal placental tissues in 30 women with PIH and 30 with normal pregnancy, and the results were analyzed with independent-sample t test and Pearson correlation analysis.

RESULTSANG II level in the maternal placental tissue homogenate showed no significant difference between women with PIH and normal pregnant women (8.51+/- 4.01 vs 7.76+/-3.47 pg/ml, P>0.05), but women with PIH had significantly higher ANG II in the fetal placental tissue (11.82+/-3.92 vs 9.64+/-2.63 pg/ml, P<0.05). ANG II level was significantly higher in the maternal blood of women with PIH than in normal pregnant women (46.44+/-8.48 vs 32.43+/-5.87 pg/ml, P<0.001), but similar in the cord blood (68.83+/-8.68 vs 72.47+/-8.51 pg/ml; P>0.05). A positive correlation was indicated between the cord blood and maternal peripheral blood ANG II levels in women with PIH (r=0.7379, P<0.05).

CONCLUSIONANG II in the fetal placental tissue is elevated, and the cord blood and maternal peripheral blood ANG II levels are positively correlated in women with PIH.

Angiotensin II ; blood ; isolation & purification ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Female ; Fetal Blood ; chemistry ; Fetal Growth Retardation ; blood ; Humans ; Hypertension, Pregnancy-Induced ; blood ; Placenta ; chemistry ; Pregnancy

OBJECTIVETo study the changes in gut microbiota and serum D-lactate level and their significance in children with recurrent pneumonia.

METHODSThe stool and blood samples were collected from 30 children with recurrent pneumonia (recurrent group), 30 children with acute pneumonia (acute group), and 15 children receiving surgical operation (surgery group). The 16S rRNA fluorescent quantitative polymerase chain reaction (FQ-PCR) was applied to determine the numbers of Bifidobacterium and Escherichia coli in stool samples, and the ratio between the logarithmic values of the numbers of Bifidobacterium and Escherichia coli (B/E value) was calculated. The serum D-lactate level was measured, and correlation analysis was performed.

RESULTSThe recurrent group had a significantly lower number of Bifidobacterium and a significantly lower B/E value than the acute group and the surgery group (P<0.05), as well as a significantly higher number of Escherichia coli than the surgery group (P<0.05). There was no significant difference in the number of Escherichia coli between the recurrent group and the acute group. The recurrent group had a significantly higher serum D-lactate level than the acute group and the surgery group (P<0.05). In the recurrent group, B/E value was negatively correlated with serum D-lactate level (r=-0.539, P<0.05).

CONCLUSIONSChildren with recurrent pneumonia may have biological and mechanical barrier damage in the intestinal mucosa.

Bacteria ; classification ; genetics ; isolation & purification ; Bifidobacterium ; genetics ; isolation & purification ; Child ; Child, Preschool ; Escherichia coli ; genetics ; isolation & purification ; Feces ; microbiology ; Female ; Gastrointestinal Microbiome ; Humans ; Lactates ; blood ; Pneumonia ; blood ; microbiology ; pathology ; Recurrence

The study was purposed to analyze the relationship between the content of T-cell receptor excision DNA circles (TREC) and bcr-abl mRNA levels in CML patients and to evaluate the prognostic significance of recent thymic output function detection in patients with chronic myelogenous leukemia (CML). Quantitative detection of TREC and bcr-abl fusion gene transcripts in peripheral blood from 15 CML patients were preformed by real-time PCR. The change of bcr-abl levels in 6 patients was followed-up for two years. The results showed that there was no significant correlation between TREC and bcr-abl mRNA levels in peripheral blood from CML patients for the first attack. Patients who had higher TREC at diagnosis had a larger reduction of bcr-abl after 2 years of follow-up. While out of 2 patients who underwent haemopoietic stem cell transplantation (HSCT), one patient with higher level of TREC before transplantation was confirmed to express undetectable level of TREC by three consecutive detections after transplantation, other one patient was identified to express low level of bcr-abl. It is concluded that high thymic output function in CML patients can be beneficial for killing the residual CML cells.
Adolescent ; Adult ; Aged ; Female ; Fusion Proteins, bcr-abl ; biosynthesis ; genetics ; Gene Rearrangement ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; Male ; Middle Aged ; Proto-Oncogene Proteins c-abl ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcr ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Antigen, T-Cell ; analysis ; immunology ; T-Lymphocytes ; chemistry ; immunology ; Thymus Gland ; immunology

OBJECTIVEIn order to fully understand hepatitis c virus (HCV) genotype 3b, 1a, 2b and 6a infection in China, We built HCV 5'-noncoding region (5'-NCR) of different genotypes and subtypes.

METHODSThe classification HCV into variable genotypes (subtypes) was carried on by programs A, B and C A. Using a combination of three restriction endonuclease BHH' (BsrB I, Hae II, Hinf I) digestions at the same time. The distinct genotypes were classified into 5 groups: genotype 1 (1a, 1b), 6a, 2 (2a, 2b), genotype 3 (3a, 3b), genotype4 (4a). B. With regard to genotype 1, we could distinguish subtype 1a from 1b using BstU I digestion. C. Using restriction endonuclease Hae III, genotype 2a, 2b, 3b, 4a, 6a are differentiated respectively.

RESULTS(1) HCV genotype 1a, 1b, 2a, 2b, 3a, 3b, 4a, 6a are fully discriminated by comparison with the genotypes regular samples. (2) Of the 93 patients, HCV genotype distribution in China was 66.67% for 1b, 18.28% for 2a, 3.23% for 1b/2b, 3b, 2b respectively. 2.15% for 2a/2b, 1b/2a respectively. 1.08% for 1a.

CONCLUSIONThis research indicated that adoption of HCV 5'-NCR A B C restriction endonuclease digestions techniques, might be sensitive and efficient to detect HCV and discriminate HCV genotype (subtypes) 1a to 6a.

5' Untranslated Regions ; chemistry ; DNA Restriction Enzymes ; Genotype ; Hepacivirus ; classification ; genetics ; RNA, Viral ; analysis

To identify the inhibitor of glutathione S-transferase (GST), a high-throughput screening method was established in a 384-well microplate with total 35 microL volume, and the absorbance at 340 nm is detected. The concentrations of substrates, CDNB and GST were determined by chromatometry. The optimal enzyme kinetics reaction time and temperature are 2 h and 30 degrees C , respectively. The established model was evaluated by NaOCl, a known GST inhibitor, and the parameter Z' was 0.77, which showed a high feasibility and stability of the assay. A total of 31,098 compounds were screened, of which 4 compounds were shown to inhibit GST activity, high inhibiting activity for their IC50 of GST inhibition was 3.94, 4.05, 74.85, and 77.41 mg x L(-1), separately. The results indicated that the colorimetric method by using CDNB and GSH as substrate is stable, sensitive, reproducible and also suitable for high throughput screening.
Dinitrochlorobenzene ; chemistry ; Drug Evaluation, Preclinical ; methods ; Enzyme Inhibitors ; analysis ; Glutathione ; chemistry ; Glutathione Transferase ; antagonists & inhibitors ; Substrate Specificity

BACKGROUND: Studies have shown that bone marrow mesenchymal stem cells have the potential of differentiation into alveolar epithelial cells in vitro, but so far no study has indicated that adipose-derived mesenchymal stem cells (ADSCs) can be differentiated into alveolar epithelial cells through long-term Transwell co-culture. OBJECTIVE: To observe whether rat lung epithelial-T-antigen negative cell lines (RLE-6TN) can induce rat ADSCs to differentiate into type II alveolar epithelial cells by long-term Transwell co-culture. METHODS: Three SPF health female Sprague-Dawley rats were used as donors to separate, extract, culture and identity ADSCs. The experimental group was subjected to the Transwell co-culture of ADSCs and RLE-6TN, while the control group was subjected to the culture of ADSCs alone. The morphological changes of ADSCs were observed by the inverted phase contrast microscope at 21 days after co-culture. Immunofluorescence staining using surfactant protein C (SP-C) was performed on the co-cultured ADSCs. The fluorescence staining was observed using the inverted fluorescence microscope. Integral optical density (IOD) analysis was conducted by Image pro plus 6.0 software. RESULTS AND CONCLUSION: RLE-6TN cells were identified by fluorescence staining with stable expression of SP-C protein (red fluorescence) in the experimental group, and there was no red fluorescence in the control group. After 21-day co-culture, the cell shape in the experimental group was transformed from the long spindle shape into oval or polygon shape gradually, while the cell shape in the control group remained fibroblast-like. These results show that RLE-6TN can induce ADSCs to differentiate into type II alveolar epithelial cells after a long-term (21 days) co-culture.

Objective To evaluate the role of cerebral state index(CSI),burst suppression (BS)and electromyograph(EMG)in monitoring coma/consciousness depth and damage degree of brain. Methods CSM was done in 50 cases with brain injury and coma to analyze its relation with physical reflection,auditory evoked potential(AEP),Glasgow coma score(GCS)and Glasgow outcome scale (GOS).Results As scale range meaning from consciousness to deep coma and to brain death,CSI 0- 100 was positively correlated with coma depth,coma score of GCS and physical reflection.CSI changes under invariable ache stimulation in combination with BS and EMG can accurately estimate prognosis and quantify changes of brain function.Conclusions The quantifiable digit of coma/consciousness depth and damage degree in brain function by CSM can attain real time judgment of dynamic evolvement course of coma and objective guide clinical therapy and assure prognosis,as will change absolutely scoring coma/ consciousness depth and prognosis under current state of artificial diversity and lacking objective evi- dences.