Objective To evaluate the effect of lovastatin on shedding of heparan sulfate proteoglycan (HSPG) and syndecan-1 (SDC-1) in the lung tissues of rats with sepsis-induced acute lung injury.Methods One-hundred and twenty male Wistar rats aged 8-12 weeks,weighing 325-425 g,were randomly divided into 3 groups (n =40 each) using a random number table:sham operation group (group S),cecal ligation and puncture (CLP) group and lovastatin group (group L).Lovastatin 4 mg/kg was injected once a day for 5 consecutive days in S and L groups,while the equal volume of 0.5％ CMC (the solvent) was given in CLP group.Sepsis was produced by CLP on 5th day of administration in CLP and L groups.The left lung was lavaged at 24 h after operation.The broncho-alveolar lavage fluid (BALF) was collected for determination of protein concentrations,white blood cell (WBC) count and percentage of neutrophils.Blood samples were collected for determination of the concentrations of HSPG and SDC-1 in serum (by ELISA).Evans blue was injected at 24 h after operation in the remaining 20 rats of each group.The lungs were removed for examination of the pathological changes and for measurement of HSPG and SDC-1 mRNA and protein expression (using Western blot and PCR),and Evans blue content (reflecting pulmonary capillary permeability) in the lung tissue.Results Compared with group S,the protein concentrations,WBC count and percentage of neutrophils in BALF,Evans blue content in lung tissues and the concentrations of HSPG and SDC-1 in serum were significantly increased,and HSPG and SDC-1 mRNA and protein expression was down-regulated in CLP and L groups.Compared with group CLP,the protein concentrations,WBC count and percentage of neutrophils in BALF,Evans blue content in lung tissues and the concentrations of HSPG and SDC-1 in serum were significantly decreased,and HSPG and SDC-1 mRNA and protein expression was up-regulated in group L.The pathological changes of lungs were significantly attenuated in group L as compared with group CLP.Conclusion The mechanism by which lovastatin attenuates acute lung injury induced by sepsis may be related to reduced shedding of HSPG and SDC-1 in lung tissues and improved function of pulmonary vascular endothelium in rats.

Objective To observe the protective effects of lovastatin against lung injury and the expression changes of adiponectin in the septic rats.Methods Fifty four male Wistar rats weighting 250-300g were randomly divided into the three groups:sham operation group ( group Sham) ,sepsis group ( group CLP) and lovastatin interven tion group (group LOV).Rats were injected with lovastatin (4mg/kg) or 0.5%CMC (vehicle) for five days and then subjected to CLP.At 4h,12h and 24h after operation.BALF was collected to determine the levels of TNF-αand IL-6,lung tissue was obtained to observe histopathological changes,and to detect the content of MPO and MDA,the blood sample was obtained to detect the level of adiponectin.Results In the group Sham,at 4h,12h and 24h time points,the levels of TNF-αwere (1.80 ±0.13)pg/mL,(2.04 ±0.15)pg/mL and (1.930.19)pg/mL;the levels of IL-6 were (20.56 ±0.23)pg/mL,(18.35 ±0.15) pg/mL and (21.23 ±0.20) pg/mL;the contents of MPO were (2.82 ±0.14) U/g tissue,(2.88 ±0.07) U/g tissue and (2.76 ±0.18) U/g tissue;and the levels of MDA were (3.32 ±0.12)nmol/mg,(3.09 ±0.11)nmol/mg and (3.21 ±0.08)nmol/mg;the concentrations of adiponectin were (2.68 ±0.14)μg/mL,(2.80 ±0.07)μg/mL and (2.86 ±0.18)μg/mL.Compared with group Sham,both LOV group and CLP group had increased pulmonary damage:(1)the levels of TNF-α[4h,12h and 24h were (4.23 ± 0.18)pg/mL,(5.62 ±0.24)pg/mL and (5.14 ±0.10)pg/mL,t=28.41,30.98 and 36.62]and IL-6[4h,12h and 24h were (39.12 ±0.17) pg/mL,(47.25 ±0.21) pg/mL and (44.690.27) pg/mL,t =158.90,273.40 and 127.28] of the CLP group in BALF were both increased,and MPO[4h,12h and 24h were (4.85 ±0.13) U/g tissue, (6.17 ±0.08)U/g tissue and (7.84 ±0.10)U/g tissue,t=26.39,79.40 and 60.43]and MDA[4h,12h and 24h were (6.24 ±0.06)nmol/mg,(7.56 ±0.15)nmol/mg and (8.43 ±0.10)nmol/mg,t=53.31,58.86 and 90.06] concentrations in lung homogenate were raised with the decreased expression of serum adiponectin[4h,12h and 24h were (1.35 ±0.10)μg/mL,(1.17 ±0.07)μg/mL and (1.24 ±0.11)μg/mL,t=19.86,12.75 and 18.81](all P<0.05);(2) meanwhile the levels of TNF-α[4h,12h and 24h were (2.85 ±0.17) pg/mL,(3.720.13) pg/mL and (3.240.09)pg/mL,t=12.02、20.73 and 16.68]and IL-6[4h,12h and 24h were (30.75 ±0.22)pg/mL, (37.52 ±0.29)pg/mL and (32.43 ±0.26)pg/mL,t=78.42,68.29 and 83.67]in BALF of the LOV group were all increased,the contents of MPO[4h,12h and 24h were(3.59 ±0.05)U/g tissue,(4.67 ±0.11)U/g tissue and (5.33 ± 0.05)U/g tissue,t=12.03,33.63 and 33.70]and MDA[4h,12h and 24h were (4.45 ±0.10)nmol/mg,(5.01 ± 0.11)nmol/mg and (5.83 ±0.04) nmol/mg,t =17.72,30.23 and 71.75] were also increased with the serum adiponectin concentrations[4h,12h and 24h were (2.09 ±0.08)μg/mL,(2.07 ±0.05)μg/mL and (2.03 ± 0.10)μg/mL,t=8.96,20.79 and 6.30]dicreased(all P<0.05).There were less histopathological changes in the LOV group,and the levels of TNF-α(t=13.46,17.05 and 15.43),IL-6(t=73.70,64.10 and 80.12),MPO(t=22.16,27.01and 32.86) and MDA(t=37.59,42.72 and 59.13) were lower than those in CLP group,also the level of adiponectin(t=14.15,8.10 and 3.19) increased siginificantly(all P<0.05).Conclusion Administration of lovastatin could attenuate lung injury of the sepsis by down-regulate the level of TNF-αand IL-6,with reduced inflam-matory response and oxidative stress,and could upregulate the level of adiponectin in serums of rats with sepsis.

To obtain recombinant human β2-microglobulin (rhβ2M) with properties of good solubility and high purity from E. coli, prokaryotic expression conditions were optimized and protein purification was performed in this study. After testing the effect of different IPTG concentrations, temperatures and induction times on the production of rhβ2M, the optimum expression conditions were determined, i. e. joining IPTG to final concentration being 0.8 mmol/L and inducing time 6 h and at temperature of 25 degrees C. Under the optimum induction conditions, the ratio of soluble rhβ2M to soluble bacterial protein was 63.7%. After purified by Ni Sepharose 6 Fast Flow, the purity of rhβ2M achieved a greater value of 95%. Western blot analysis revealed that rhβ2M possessed the antigen property that specifically interacted with anti-β2M antibody.
Blotting, Western ; Escherichia coli ; metabolism ; Humans ; Recombinant Proteins ; biosynthesis ; Solubility ; beta 2-Microglobulin ; biosynthesis

Objective To evaluate the effects of rapamycin combined with rosiglitazone on lung injury in septic rats.Methods One hundred and twenty healthy male Wistar rats were randomly divided into 5 groups (n =6 each):sham operation group (group S); cecum ligation and punture (CLP) group; rapamycin group (group RPM) ; rosiglitazone group (group RGZ) ; rapamycin plus rosiglitazone group (group RPM + RGZ).The rats were anesthetized with intraperitoneal 10% chloral hydrate 100 mg/kg.Sepsis was induced by CLP in groups CLP,RPM,RGZ and RPM + RGZ.At 30 min before CLP,rapamycin 0.4 mg/kg was injected subcutaneously in RPM group,rosiglitazone 0.3 mg/kg was injected via the femoral vein in RGZ group,and rapamycin 0.4 mg/kg was injected subcutaneously and rosiglitazone 0.3 mg/kg was injected via the femoral vein in group RPM + RGZ.While the equal volume of normal saline was given instead in CLP group.Six rats were sacrificed at 2,6,24 and 48 h after CLP in each group,and lungs were removed and cut into sections which were stained with haematoxylin and eosin and examined under microscope.The pathological changes of lungs were scored.The myeloperoxidase (MPO) activity and signal transducer and activator of transcription 3 (STAT3)-DNA binding activity in lung tissues were measured.Results Compared with group S,the pathological scores,MPO activity and STAT3-DNA binding activity were significantly increased in groups CLP,RPM,RGZ,RPM + RGZ (P ＜ 0.05).The pathological scores,MPO activity and STAT3-DNA binding activity were significantly lower in groups RPM,RGZ and RPM +RGZ than in group CLP,and in group RPM + RGZ than in groups RPM and RGZ (P ＜ 0.05).Conclusion Rapamycin combined with rosiglitazone offers additional benefit to attenuating lung injury induced by sepsis over rapamycin or rosiglitazone alone,and inhibition of activation of STAT3 pathway is involved in the mechanism.

Objective To evaluate the effect of dexmedetomidine on the expression of 8-Oxoguanine DNA glycosylase 1 (OGG1) mRNA in rat hippocampal neurons subjected to oxygen-glucose deprivation/reoxygenation and restoration (OGD/R).Methods Hippocampal neurons isolated from pathogen-free neonatal Sprague-Dawley rats born within 3 days,were cultured primarily and seeded in 96-well plates (100 μl/well) or 6-well plates (2 ml/well) at the density of 1 × 106 cells/ml.The cells were randomly divided into 5 groups (n=30 each):control group (group C),group OGD/R,and different concentrations of dexmedetomidine groups (DEX1-3 groups).The cells were cultured in normal culture medium in group C and the cells were subjected to OGD/R in the other groups.In DEX1-3 groups,dexmedetomidine with the final concentrations of 0.1,1.0 and 10.0μmol/L were added,respetively,at 2h before OGD.At 24h of restoration,hippocampal neurons were stained with haematoxylin and eosin (H.E) for examination of pathological changes,the cell survival rate was detected by MTT method,the activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) were detected by colorimetric method,and the expression of OGG1 mRNA was detected by RT-PCR.Results The pathological changes of neurons were obvious in group OGD/R,and the pathological changes of neurons were significantly mitigated in DEX1,DEX2 and DEX3 groups.Compared with group C,the cell survival rate and SOD activity were significantly decreased,MDA content was increased,and the expression of OGG1 mRNA was down-regulated in OGD/R,DEX1,DEX2 and DEX3 groups (P ＜ 0.05).Compared with group OGD/R,the cell survival rate and SOD activity were significantly increased,MDA content was decreased,and the expression of OGG1 mRNA was up-regulated in DEX1,DEX2 and DEX3 groups (P ＜ 0.05).There was no significant difference in the indices mentioned above between DEX1,DEX2 and DEX3 groups (P ＞ 0.05).Conclusion Dexmedetomidine may protect hippocampal neurons against oxidative stress injury by up-regulating the expression of OGG1 mRNA in rat hippocampal neurons subjected to OGD/R.

Objective To evaluate the effects of adiponectin on the mitochondrial damage in septic rats with lung injury .Methods Sixty male Wistar rats ,aged 7-8 weeks ,weighing 180-220 g ,were randomly divided into 3 groups (n=20 each) using a random number table :sham operation group (S group) ,sepsis group (SEP group) and adiponectin group (APN group) .The animals were anesthetized with intraperitoneal 2% pentobarbital sodium 50 mg/kg .Sepsis was produced by cecum ligation and puncture (CLP) .In APN groups ,gene recombinant adiponectin 6 mg/kg was injected intraperitoneally at 12 h after CLP .At 24 h after the operation ,10 rats from each group were chosen and sacrificed ,and lungs were removed for detection of interleukin-6 (IL-6 ) and high-mobility group box-1 (HMGB1 ) contents . The mitochondria of lung samples were isolated for measurement of the malondialdehyde (MDA ) content and degree of mitochondrial swelling and mitochondrial activity . The survival rates within 7 days after operation were recorded in the left 10 rats in each group .Results Compared with S group , IL-6 and HMGB1 contents in lung tissues and MDA content in the mitochondria were significantly increased ,the degree of mitochondrial swelling was increased , and the mitochondrial activity and survival rates within 7 days after operation were decreased in SEP group ( P<0.05 ) . Compared with SEP group , IL-6 and HMGB1 contents in lung tissues and MDA content in the mitochondria were significantly decreased ,the degree of mitochondrial swelling was reduced ,and the mitochondrial activity and survival rates within 7 days after operation were increased in APN groups ( P< 0.05 ) .Conclusion Adiponectin can attenuate lung injury and raise the survival rates in septic rats ,and reduction of mitochondrial damage in lung tissues is involved in the mechanism .

OBJECTIVETo detect potential mutation of COL2A1 gene in two children suspected for Kniest dysplasia.

METHODSThe 54 exons and splicing regions of the COL2A1 gene were amplified with PCR and the product was subjected to direct sequencing.

RESULTSA missense mutation (c.905C>T, p.Ala302Val) was found in the coding region of the COL2A1 gene, which has been previously reported in abroad. The patients appeared to have short trunk dwarfism, enlarged joints and midface hypoplasia.

CONCLUSIONThe probands are the first cases of Kniest dysplasia described in China, and so was the p.Ala302Val mutation.

Base Sequence ; Child, Preschool ; China ; Cleft Palate ; genetics ; Collagen Diseases ; genetics ; Collagen Type II ; genetics ; Dwarfism ; genetics ; Exons ; Face ; abnormalities ; Humans ; Hyaline Membrane Disease ; genetics ; Male ; Molecular Sequence Data ; Mutation, Missense ; Open Reading Frames ; Osteochondrodysplasias ; genetics ; RNA Splicing

OBJECTIVECockayne syndrome is a rare disease and difficult to be recognized. This study aimed to expand the knowledge of the clinical and molecular characteristics of the children with Cockayne syndrome (CS).

METHODClinical data of two siblings with classic CS of Guangzhou Women and Children's Medical Center from July 2013 to November 2014 were obtained and analyzed. The whole DNA of peripheral blood was collected from two CS siblings and their parents. Amplification of all exons and adjacent introns for ERCC6 gene was conducted using PCR, and measurement of reaction product was performed to find mutation sites by two-way sequencing.

RESULTTwo affected siblings were males, and came from unconsanguineous parents, 7 years and 5 months old and 4 years and 8 months old, respectively. They were in treatment because of developmental and mental retardation for years. When they were younger than one year of age, their heights and weight were within normal limits. However, poor growth of height and weight and psychomotor retardation appeared after one and a half years of age, as well as skin and eye sensitivity to sunshine, hearing impairment, optic nerve atrophy, microcephaly, and deep-set eyes. The proband's height was 90.8 cm, and weight 9.1 kg, head circumference 41 cm, and chest circumference 44 cm when he was taken to hospital. The elder brother of the proband had a height of 92 cm, weight 11.2 kg, head circumference 41 cm, and chest circumference 44 cm when he was taken to hospital. When the proband was four and a half years old, ventricular enlargement, hypomyelination, and brain atrophy were detected for his elder brother at 7 years of age by cranial MRI. MRS imaging indicated that damages occurred at the left and right sides of dorsal thalamus, lobus insularis, along with the left half circle of central neurons. Symmetrical calcification on bilateral basal ganglia was found on the brain CT scan. Pathogenic compound heterozygous c. 1357C > T (p.Arg453Ter) and c. 1607T > G (p.Leu536Trp) mutations of ERCC6 gene were identified in the two siblings which were separately inherited from their unaffected parents.

CONCLUSIONCS children are usually normal at birth, however, they have severe clinical characteristics such as poor growth, psychomotor retardation, cerebral injury, microcephalus, deep-set eyes, and skin sensitivity to sunshine. ERCC6 gene mutation usually occurs, and it is easy to misdiagnose CS as cerebral palsy, primary microcephaly, and so on.

Asian Continental Ancestry Group ; Child ; Child, Preschool ; Cockayne Syndrome ; genetics ; DNA Helicases ; genetics ; DNA Mutational Analysis ; DNA Repair Enzymes ; genetics ; Exons ; Heterozygote ; Humans ; Magnetic Resonance Imaging ; Male ; Mutation ; Poly-ADP-Ribose Binding Proteins ; Polymerase Chain Reaction ; Siblings

Objective:To construct a standardized training program for full-time nursing postgraduate students, and to provide a basis for the training the high-quality nursing talents in hospitals.Methods:Through literature review and demi structured interview, preliminarily develop the training program was established from January to March 2021. Through two rounds of expert consultations of 16 experts, the standardized training program for nursing postgraduates was formulated.Results:The recovery rate of the two rounds were both 16/16. The expert authority coefficients of the two rounds of consultation were 0.819, respectively. The Kendall′s coordination coefficients of the two rounds of consultation were 0.329 and 0.334, respectively, and the difference was statistically significant ( P< 0.01). Finally, a standardized training program for entry-level nursing postgraduates was formed, which included five parts: training objects and quality requirements, training methods, training objectives, training contents and evaluation, with a total of 225 items. Conclusions:The standardized training program for nursing postgraduates formed is reliable, scientific and practical, which provides guidance and reference for the training and use of highly educated clinical nursing talents in hospitals.