Acupuncture Therapy ; Adolescent ; Craniocerebral Trauma ; complications ; Humans ; Male ; Optic Nerve Injuries ; etiology ; physiopathology ; therapy ; Vision, Ocular

Objective To detect anti-heterogeneous nuclear ribonucleoprotein A2 (hnRNP A2)/RA33 antibody by ELISA with the purified recombinant hnRNP A2 antigen. Methods The serum of 179 patients with RA, 141 patients with SLE, 97 patients with other diffused rheumatic diseases, 30 patients with seronegative spondyloarthropathies, 10 patients with osteoarthritis, 59 patients with arthralgia/arthritis and 40 controls were detected. In addition, clinical characters and laboratory indexes were compared to study the significance of anti-hnRNP A2/RA33 antibody in RA. Results The sensitivity and specificity of anti-hnRNP A2/RA33 antibody in RA were 36.9% and 87.1%. The positive rates of anti-hnRNP A2/RA33 antibody in SLE, other CTD, seronegative spondyloarthropathies and OA were 19.2%, 7.2%, 6.8% and 0. The positive rate of anti-hnRNP A2/RA33 antibody was 43.3% in early RA patients. Conclusion Detection of anti- hnRNP A2/RA33 antibody with purified recombinant hnRNP A2 antigen is a reliable method for early diagnosis of RA.

Objective To search specific biomarkers of pathogenic bacteria in patients with sepsis so as to guide early using rationally antibiotic treatment.Methods Prospective survey of 147 patients with sepsis in ICU was carried out from Jan 2012 to Mar 2015.When patients blood culture was positive, clinical data including age, gender, vital signs, blood and, urine routine examination, DIC, blood biochemistry, c-reactive protein (CRP), procalcitonin (PCT), microbial detection, etc were recorded.Cultured blood samples were from central venous catheter and peripheral vessel.ELISA method was employed to detect soluble toll-like receptor 2 ( sTLR2 ) and interleukin-8 ( IL-8 ) , and the Acute Physiology and Chronic Health Evaluation Ⅱ( APACHEⅡ score ) was calculated.The chi-square test and analysis of variance were performed where necessary.Receiver operating characteristic ( ROC ) curves were used to calculate cut-points ( CP ) and area under the curve ( AUC) .Results According to the results of blood culture, patients were divided into three groups:GP group [ gram-positive bacteria ( G+) group];GN group [ gram-negative bacteria ( G-) group];FG group ( fungi group) .There were no significantly statistical differences in age, APACHEⅡ score, vital signs and markers of inflammation among three groups (P>0.05).Gram negative pathogenic bacterium was the most common microbe.Compared with GN group, the level of sTLR2 in the GP group was obviously higher ( P=0.000); but there was no significant difference in sTLR2 level between GP group and FG group (P=0.187). The amount of (1, 3) -beta glucan in the FG group was significantly higher than that in the GP group ( P=0.000).The sTLR2 level in FG group was obviously higher than that in the GN group (P=0.000).There were no significantly statistical differences in PCT, CRP and IL-8 among the three groups (P>0.05).For the diagnosis of gram negative bacteria infection, sTLR2 area under the curve was 0.768, and the sensitivity and specificity were 88.90%and 59.60%, respectively and the best cut-off point was 8.083 pg/mL.Namely, the diagnosis of gram negative bacteria infection was less likely, when level of sTLR2 was higher than 8.083 pg/mL.The markers of PCT, CRP, (1, 3) -beta glucan and IL-8 were less valuable for the diagnosis of Gram negative bacteria infection because the area under the curve was less than 0.5.Conclusions The combination of inflammatory indicators such as sTLR2 and (1, 3) -beta glucan etc, can imply the kind of pathogenic microorganisms partly.

Objective To observe the effect of Astragalus injection on the expressions of inflammatory cytokines in human primary macrophages stimulated by lipoteichoic acid (LTA) and lipopolysaccharide (LPS),and investigate its effects on inflammatory reactions of Gram-positive (G+) and Gram-negative (G-) bacteria sepsis and its mechanisms.Methods Percoll density gradient centrifugation was used to isolate the human peripheral blood mononuclear cells,then they were purified by immune Anti-Biotin Microbeads with magnetic character and under the induction of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF),the cells were cultivated for 12 days in vitro,eventually the human monocyte-derived macrophage was formed.The cultured human macrophages were inoculated in 96-well plates (each group 3 wells) and 6-well plates (each group 3 wells).The cells were divided into control group (200 μL DMEM added in each well),LTA 1 mg/L group,LPS 0.1 mg/L group and low astragalus injection (0.1 mg/L) and high astragalus injection (0.2 mg/L) dose groups.After the incubator plates were put in an incubator for 24 hours,the protein content of IL-8 and IL-10 in supernatant were detected by enzymelinked immunosorbent assay (ELISA),and the mRNA expression levels of IL-8 and IL-10 were detected by real time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR).Results LTA and LPS all can obviously up-regulate the expression levels of pro-inflammatory factor IL-8 and anti-inflammatory factor IL-10 of macrophage.The expressions of IL-8 and IL-10 protein and mRNA in LTA group and LPS group were significantly higher than those in control group after cuhure for 8 hours and 24 hours,the degrees of increment were more significantly at 24 hours [LTA stimute group:IL-8 protein (ng/L,× 103):41.57± 1.90 vs.1.58 ±0.24,IL-8 mRNA (A value):21.49±8.05 vs.1.00±0.16;IL-10 protein (ng/L):5.90±3.02 vs.2.91 ± 1.54,IL-10 mRNA (A value):1.35±0.34 vs.0.95±0.14;LPS stimute group:IL-8 protein (ng/L,× 103):345.00±22.80 vs.5.60±0.31,IL-8 mRNA (A value):29.84 ± 8.93 vs.1.00 ± 0.16,IL-10 protein (ng/L):122.37 ± 39.26 vs.44.79 ± 3.67,IL-10 mRNA (A value):7.38 ± 1.58 vs.1.35 ± 0.34,all P ＜ 0.05].The Astragalus injection could regulate LTA and LPS to stimulate the macrophage to decrease the expression levels of pro-inflammatory factor IL-8 protein and mRNA and increase the expression levels of anti-inflammatory factor IL-10 protein and mRNA in the macrophage;the changes of regulatory effect in the 24 hour-culture of Astragalus injection high dose group was the most significant [LTA stimute group:IL-8 protein (ng/L,×103):22.63±1.91 vs.41.57±1.90,IL-8 mRNA (A value):12.10±1.93 vs.21.49±8.05,IL-10 protein (ng/L):14.03±2.22 vs.5.90±3.02,IL-10 mRNA (A value):10.37±6.08 vs.1.35±0.34;LPS stimute group:IL-8 protein (ng/L,× 103):167.75 ± 19.90 vs.345.01 ±22.80,IL-8 mRNA (A value):15.61 ± 3.63 vs.29.84±8.93;IL-10 protein (ng/L):243.22±14.41 vs.122.37±39.26,IL-10 mRNA (A value):16.14±4.10 vs.7.38± 1.58,all P ＜ 0.05].Conclusions In the process of inflammatory response,the pro-inflammatory and anti-inflammatory factors co-exist simultaneously.Astragalus injection can inhibit the expression levels of pro-inflammatory factor gene and protein in the inflammatory response of G+ and G-bacteria sepsis and in the mean time,it can promote the expression levels of anti-inflammatory factor gene and protein,thus the immune mechanism of sepsis is affected,achieving the balance between pro-inflammation and anti-inflammation.

Objective To establish the HepG2 cell lines which can stably express and replicate hepatitis 13 virus (HBV).Methods One point two X unit length of HBV genome was cloned intn SalⅠsite of the eukaryotic expression vector pREP10 to construct the recombinant plasmid pREP-HBV. Human hepatoblastoma cell HepG2 was transfected with pREP-HBV by Lipofectamine 2000 and seh,cted by bygromycin at the concentration of 250?g/mL.HBsAg and HBeAg were monitored by enzyme linked immunosorbent assay (ELISA)kits.H13V particles presemed in supernatant were ex- amincd by electronic microscopy.DNA isolated from intracellular HBV core particles was analyzed by Southbern blot using HBV-specific probe.Results The recombinant vector pREP HBV containing 1.2?unit length of HBV DNA was constructed successfully.After transfection of pREP-HBV to HepG2 cells and consistently cultured in hygromycin selective medium.5 drug-resistant cell lines, RHBV1-RHBV5.were established,and all of them could stably express HBsAgand HBeAg.South ern blot analysis revealed that HBV could replicate in all cell lines,as confirmed by the presence of replicateintermediatc DNA in intracellular HBV core particles.Clustered 42 nm Dane particles as well as 22-26 nm spherical H13sAg particles in condensed cuhure supernatant were visualized by elec tronic microsopic analysis.Conclusion HepG2 ceil lines in which HBV can replicate and express specific antigens are successfully established.Up to now,the cells have been passaged every three days for 50 times.

ObjectiveTo discuss the experimental method and the mechanisms on treating diseases by acupuncturing the ST36(Zusanli).MethodsUsing Positron Emission Tomography(PET) and functional Magnetic Resonance Imaging(fMRI) to obtain the experimental data about glycometabolism and cerebral blood stream,using SPM and ROI image-analytical method to obtain the visual experimental evidence when acupuncturing the ST36. ResultsThere are certain increases of glycometabolism and cerebral blood stream in ipsilateral hypothalamus and bilateral temporal lobe, when acupuncturing the ST36. Conclusions Acupuncturing the ST36 can lead to the functional changes in vegetative nerve center and temporal lobe, which is close correlated with the therapeutical effects of ST36.

ObjectiveTo determine the impact of lupus flares on maternal and fetal outcomes in pregnant patients with systemic lupus erythematosus(SLE).MethodsData was obtained from 46 pregnancies of 44 pregnant women with SLE.The relationship between lupus flares and pregnant outcomes,and the risk factors for adverse maternal and fetal prognosis were analyzed.T-test,X2 test or Fisher's exact test and Logistic regression were used for statistical analysis.Results① Lupus flares occurred in19(41％)pregnancies(group A) and stable lupus disease was observed in 27(59％) pregnancies(group B) during pregnancy.Compared to pregnancies in patients with stable lupus disease at the conception(n=32),pregnancies in patients with unstable lupus disease at the conception(n=8) had higher lupus flare during pregnancy( 100％ vs 16％,P＜0.05).(②) The common manifestations of lupus flares during pregnancy were lupus nephritis (LN) (11 cases),skin rashes (10 cases),arthritis (7 cases),and the common complication was infection ( 11 cases).(③) The incidence of premature labor,fetal growth retardation (FGR) and fetal loss in group A was 42％,47％ and 26％ respectively,which was significantly higher than that of the group B (7％,15％ and 0 respectively)(P＜0.05).There was no difference in the incidence of preeclampsia,fetal distress and neonatal asphyxia between the two groups ( 16％ vs 7％,16％ vs 19％,5％ vs O,respectively,P＞0.05).The incidence of premature labor and FGR in patients with active LN was higher than that of patients without active LN (55％ vs 11％,64％ vs 17％,respectively,P＜0.05).(④)The binary Logistic regression analysis showed that renal impairment,hypocomplementemia,aPL and serum urea nitrogen level were independent risk factors for premature delivery,FGR,fetal loss and fetal distress.Conclusion(①) Lupus flares during pregnancy increase the incidence of premature labor,FGR and fetal loss.Active LN during pregnancy can increase the incidence of premature labor and FGR.② Renal impairment,hypocomplementemia,aPL and serum urea nitrogen level are associated with adverse fetal outcomes in pregnant patients with SLE.

OBJECTIVEThis study aims to investigate the expression levels of serum and urinary vascular endothelial growth factor-A (VEGF-A) and epidermal growth factor-like domain 7 (EGFL7) in proliferating infantile hemangioma patients under propranolol treatment.

METHODSPropranolol (0.5-2 mg x kg(-1)) was orally administered to 30 infants every day for 4-8 months. The Achauer method was used to measure the tumor radius and thus evaluate the clinical curative effects of the treatment. Enzyme-linked immunosorbent assay was used to measure the serum and urinary concentrations of VEGF-A and EGFL7 at 0, 4, and 12 weeks after the treatment.

RESULTSThe treatment response was excellent in 2 patients, good in 11, moderate in 14, and poor in 3. Serum VEGF-A (335.692 pg x mL(-1) ± 136.146 pg x mL(-1)) was high before the treatment and then significantly decreased after 4 weeks (264.853 pg x mL(-1) ± 122.120 pg x mL(-1)) and 12 weeks (211.345 pg x mL(-1) ± 104.035 pg x mL(-1)) of treatment (P < 0.05). Urinary VEGF-A (76.234 pg x mL(-1) ± 24.169 pg x mL(-1)) was high before the treatment and then significantly decreased after four weeks (56.454 pg x mL(-1) ± l6.111 pg x mL(-1)) and twelve weeks (34.728 pg x mL(-1)) ± 12.656 pg x mL(-1)) of treatment (P < 0.05). Serum and urinary EGFL7 also decreased after the treatment, showing a positive relationship with VEGF-A.

CONCLUSIONPropranolol can be safely and effectively used to treat proliferating infantile hemangiomas. This treatment can reduce the peripheral serum and urinary concentrations of VEGF-A and EGFL7 in affected children.

Objective To explore the clinical value and typing characteristic of prenatal ultrasonography for fetal cystic adenomatoid malformation of the lung(CAML).Methods Ultrasonographic features and typing of fetal CAML in 41 cases detected by prenatal ultrasonography were analyzed retrospectively.All cases were followed up until to the induction of labor or birth.Results (1)Site of tumor: in the 41 cases,there were 22 cases on the left side,15 on the right side and 4 on bilateral.(2)Typing diagnosis of prenatal ultrasonography: three cases were CAML type Ⅰ,14 were type Ⅱ,and 24 cases were type Ⅲ.(3)Pathology diagnosis: thirty two cases were induced abortion.CAML was confirmed in 29 cases by autopsy and the classifications were consistent with the prenatal diagnosis of ultrasonography.Three pulmonary sequestration cases were prenatally misdiagnosed as CAML type Ⅲ by ultrasonography.(4)The other nine cases were followed up to birth.Three masses decreased gradually and then disappeared.Six newborns were confirmed as CAML by CT.Their typings were consistent with the postnatal diagnosis.(5)The diagnostic accuracy rate of prenatal ultrasonography for CAML was 92.7%(38/41).Its misdiagnostic rate was 7.3%(3/41).Conclusions Prenatal ultrasonography has a high accuracy rate for the diagnosis and classification of CAML and is the first choice to detect CAML early in pregnancy.It has an important clinical value.Pulmonary sequestration should be distinguished from CAML type Ⅲ because they tend to be confused.

Objective To evaluate the clinical efficacy of propranolol in treating proliferating infantile haemangiomas,and to measure the expression levels of vascular endothelial growth factor-A (VEGF-A) and hypoxiainducible factor 1α (HIF-1α) in sera and urine of patients during the treatment.Methods Thirty infants with proliferating haemangiomas were treated with propranolol at doses of 0.5-2 mg/kg per day.The radius of haemangiomas was measured,and blood and urine samples were obtained from these patients before,and at 4 and 12 weeks after the beginning of treatment.Clinical efficacy was estimated according to a four-graded scale as well as the feedback from parents of these patients.Enzyme-linked immunosorbent assay (ELISA) was performed to determine the serum and urine concentrations of VEGF-A and HIF-1α.Thirty check-up infants collected from the Department of Child Health Care served as the healthy controls.Statistical analysis was done by two-way analysis of variance followed by the least significant difference (LSD) test.Results After 12 weeks of treatment,clinical response was excellent in 2 patients,good in 11,moderate in 14,and poor in 3.The serum levels of VEGF-A and HIF-1α were (268.174 ± 95.056) μg/L and (10.809 ± 1.686) mg/L respectively in the control group,sequentially decreased in the patients from baseline to 4 and 12 weeks after the beginning of treatment (VEGF-A:(385.692 ± 136.146) vs.(264.853 ± 122.12) vs.(211.345 ± 104.035) μg/L; HIF-1α:(31.462 ± 7.458) vs.(21.454 ± 5.489) vs.(12.052 ± 3.623) mg/L).The trend in expression changes of VEGF-A and HIF-1α in urine samples was similar to that in blood samples in these patients.Positive correlation was observed between the expression level of VEGF-A and HIF-1α in sera (r=0.730,P＜ 0.05) and urine (r=0.667,P＜ 0.05) of these patients.Moreover,the levels of serum VEGF-A,urine VEGF-A,serum HIF-1α and urine HIF-1α were all negatively correlated with the time course following propranolol administration (r =-0.390,-0.689,-0.806,-0.683,P ＜ 0.05,0.01,0.05,0.01 respectively).Conclusion Propranolol is effective for the treatment of proliferating infantile haemangiomas,likely by reducing serum and urinary concentrations of VEGF-A and HIF-lα in children.