Objective To evaluate a serf-designed diagnostic protoeol which can early detect a femoral neck fracture for patients with a femoral shaft fracture. Methods From September 2005 to June 2007, a self-developed protocol was used to detect an ipsilateral femoral neck fracture for all the patients with femoral shaft fracture who had sought treatment in our department. This protocol consisted of anteroposterior plain radiography of internal rotator, intraoperative fluoroscopy of the hip, a fine (2 mm) cut computed to-mographic scan through the femoral neck, postoperative anteroposterior and lateral plain radiography of the hip in the operating room prior to awakening the patient, at the time of follow-up anteroposterior and lateral plain radiography of the hip in the presence of hip pain. The diagnostic effects of the protocol were compared with those of conventional diagnosis used for all the patients with femoral shaft fracture who had sought treatment in our department from September 2003 to August 2005. A chi-square analysis comparing the protocol group (September 2005 to June 2007) and the non-protocol group (September 2003 to August 2005) was used to assess the early and delayed diagnosis rates for an associated ipsilateral femoral neck fracture. Results The earlydiagnosis rate of an associated femoral neck fracture by the protocol was 93.8%, markedly higher than that by conventional method (46.2%), with statistically significant difference (χ2 = 4.069, P =0.044). Conclu-sion In presence of a femoral shaft fracture. this protocol consisting of plain radiography of intemal rotator, intraoperative fluoroscopy of the hip, fine cut computed tomographic scan of the femoral neck, postoperative plain radiography of the hip, and follow-up plain radiography of the hip in the presence of hip pain, may sig-nificantly improve the diagnostic rate of an associated femoral neck fracture.

In order to obtain liposomes with properties of heptocyte-specificity and pH-sensitivity,four galactosylated derivatives were synthesized. A series of liposomes were prepared by mixing the galactosylated derivatives with DC-chol/DOPE respectively. The liposome 18-gal was proven to have favorable gene transfer efficiency to human hepatoma HepG2 cells, which was significantly inhibited in the presence of galactose solution, indicating that the liposomal transfection activity was mediated by asialoglycoprotein receptors. The liposome showed prominent pH-sensitivity and low cytotoxicity. Its optimum gene transfer conditions were also determined. The results showed that the liposome may be developed as a potential hepatocyte targeting pH sensitive delivery system for nucleic acid drugs.

Objective To evaluate the treatment of current status and prognosis in childhood APL with APL2008 ,which was administrated since 2008 in our center .Methods A total of 43 children with newly diagnosed APL between 2008 to 2014 were studied retrospectively .Treatment options and current status were summarized from 28 patients who received APL2008 therapy . Results Studied 43 patients were at median age of 8 years and 4 months ,with 28 boys and 15 girls .The main clinical manifestations were infection ,anemia ,bleeding ,fever ,hepatomegaly ,splenomegaly and lymphadenopathy .The proportions of low ,intermediate and high risk groups were 27 .9% ,48 .8% and 23 .3% ,respectively .Eleven cases could be diagnosed as DIC .Bone marrow morphology showed abnormal elevation of promyelocyte .37 patients had distinctive immunophenotype such as frequent expression of CD33 , CD117 and MPO .PML/RARαfusion gene positive rate was 100% in 43 children and cytogenetic analysis were positive in 37 cases , of which specific genetic lesion in APL cells with t (15 ;17)(q22 ;q12) was found in 28 cases ,and karyotypes was found in 9 cases as infrequent chromosomal abnormalities .In 43 patients ,4 cases were early dead from intracranial hemorrhage at early stage ,and 11 cases were given up early .There were only 2 cases dead ,2 cases relapsed and 1 case lost among 28 APL children ,which enabled ef‐ficacy analysis possible .96 .4% of these 28 cases achieved HCR .The 2 year Kaplan Meier estimates of OS and EFS were 85 .9% ± 7 .6% and 80 .4% ± 8 .8% .But OS and EFS would be 94 .7% ± 5 .1% and 88 .9% ± 7 .4% if 3 patients who had non standard treat‐ment were excluded .Conclusion Childhood APL were characterized by anemia ,bleeding ,fever and infiltration .APL′s coincidence rate between PML/RARa fusion gene and morphology ,immunology and cytogenetics were 95 .3% ,90 .2% and 86 .5% ,respective‐ly .APL2008 significantly improved the prognosis of APL .

Objective To express and purify the epitope peptide of human melanin-concentrating hormone receptor 1, and to evaluate its performance in the detection of autoantibodies in vitiligo patients. Methods The target gene encoding the epitope peptide of human melanin-concentrating hormone receptor 1 was synthesized, cloned to prokaryotic expression vector pGEX-4T-2 which was then transferred to E. coli BL21. The protein expression was induced by isopropy-β-D-thiogalactoside (IPTG) and identified with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. Blocking ELISA was carried out with membrane proteins extracted from melanocytes as the blocking antigen. The antigenicity of the peptide was detected in sera from 100 patients with progressive vitiligo and 30 healthy human controls. Results The recombinant expression vector was successfully constructed, and the target protein was successfully expressed in E.coli, which was evidenced by SDS-PAGE and Western blot. With the glutathione S-transferase (GST) purification kit, the purity of the recombinant protein reached 100% when the sampling weight was less than 0.625 μg.The binding of the target protein with serum IgG antibodies from vitiligo patients could be blocked by natural membrane antigen of melanocytes. Of the 100 sera from patients with progressive vitiligo, 36 were reactive with the target protein. Conclusions The epitope peptide of human melanin-concentrating hormone receptor 1 has been successfully expressed and purified. The purified protein can bind with serum IgG antibodies from vitiligo patients, and may be applied to the detection of autoantibodies against human melanin-concentrating hormone receptor 1.

Objective To discuss the relationship between serum anti-Phosphalipase A2 receptor (PLA2R) antibodies and glomerular IgG4 subclass in patients with membranous nephropathy and evaluate the diagnostic value of the two markers.Methods Patients diagnosed as membranous nephropathy from October 2011 to April 2014 in Peking Union Medical College Hospital were included and divided into IMN and SMN groups accoding to their clinical diagnosis.Serum anti-PLA2R antibodies and glomerular IgG subclasses were both detected by indirect immunofluorescence assay.Receiver operator characteristic curves were used to evaluate the diagnostic efficiency of anti-PLA2R antibodies and glomerular IgG4.Results Prevalence of serum anti-PLA2R antibodies of IMN patients was 69.5％ (41/59); prevalence of MLN patients was 4.8％ (1/21).Within the IMN group,thirty-five patients showed positive results of both serum anti-PLA2R antibodies and glomerular IgG4; Six patients were positive for serum anti-PLA2R antibodies but negative for glomerular IgG4; Seventeen patients were positive for glomerular IgG4 but negative for serum anti-PLA2R antibodies; one patient was negative for both tests.The sensitivity of serum anti-PLA2R antibody was 69.5％ and the specificity was 95.2％; the sensitivity of glomerular IgG4 was 89.8％ and the specificity was 52.3％.The sensitivity of the combined marker consisting of serum anti-PLA2R antibody and glomerular IgG4 was 59.3％ and the specificity was 100％.Four out of the six patients secondary to HBV infection,one out of the three patients secondary to Sj(o)gren syndrome,one out of the three patients secondary to malignant tumor showed positive results of serum anti-PLA2R antibodies.Conclusions Serum antiPLA2R antibodies were of high prevalence among IMN patients; the prevalence among SMN patients varied with etiologies.Results of serum anti-PLA2R antibodies and glomerular IgG4 were helpful to rule out secondary etiologies in the diagnosis of membrnous nephropathy.

BACKGROUND:Open reduction pedicle screw fixation for thoracolumbar fracture could obtain satisfactory effects, and has been extensively used. However, it has potential risk during and after repair. Minimaly invasive percutaneous pedicle screw technique minimizes the trauma and complications of soft tissue. It remains poorly understood which is better minimaly invasive percutaneous or conventional open pedicle screw fixation for the repair of thoracolumbar fracture. OBJECTIVE:To perform quality evaluation and meta-analysis on curative effect and postoperative complications of minimaly invasive percutaneous and conventional open pedicle screw fixation in the treatment of thoracolumbar fractures. METHODS:A detailed search of several electronic databases, including Cochrane Library, PubMed, WanFang, CNKI, VIP and CBM, was undertaken. Simultaneously,Chinese Journal of Orthopaedics, Chinese Journal of Orthopaedic Trauma, and Chinese Journal of Trauma were checked by hand to identify controled trials regarding minimaly invasive percutaneous and conventional open pedicle screw fixation in the treatment of thoracolumbar fractures published from inception to 2015, and the references of the included studies were checked. According to inclusion and exclusion criteria, references were screened, data were extracted and quality was evaluated by four investigators independently. Meta-analysis was conducted using RevMan 5.2 software. The quality of references of the included controled trials was assessed with CONSORT statement and some surgery clinical evaluation indexes. RESULTS AND CONCLUSION: We included 28 studies, including 25 randomized controled trials/quasi-randomized controled trials and 3 retrospective comparative studies, with 1 285 patients. Meta-analysis results demonstrated that compared with the conventional open pedicle screw, minimaly invasive percutaneous pedicle screw fixation could significantly reduce operation time, blood loss, hospital stays, postoperative complication rate and height loss (P < 0.05). No significant difference in postoperative pain grade was detected between them (P > 0.05). These results indicate that minimaly invasive percutaneous pedicle screw fixation for thoracolumbar fracture was safe and reliable, had smal trauma, less blood loss, rapid recovery, short hospital stay, and less postoperative complications. Nevertheless, methods and results of most studies are not detailed enough. We suggested reporting randomized controled trials according to related standards in order to improve the report quality and authenticity of randomized controled trials.

Objective To evaluate the effect of the difficult embryo transfer on the clinical pregnancy outcome of in vitro fertili-zation-embryo transfer(IVF-ET) .Methods There were 209 fresh cycles of difficultly transferring and 2 489 fresh cycles of easily embryo transferring between January 2011 and December 2012 .The clinical outcome was compared .Results There were statistical-ly significant differences in the catheter blood staining rates (51 .20% vs 27 .68% ,P< 0 .05) ,implantation rate(31 .14% vs 35 . 54% ,P<0 .05) ,and clinical pregnancy rate (46 .41% vs 55 .56% ,P<0 .05)between the two groups .There was no significant difference in the rates of ectopic pregnancy and miscarriage between the two groups (P>0 .05) .Conclusion Difficulty ET will in-fluence the clinical pregnancy .Therefore ,all efforts should be made to avoid the difficult transfer in order to increase the pregnant rate .

Objective To investigate the intracellular signal transduction pathways involved in the protective effect of nicotinic acid against ultraviolet B(UVB)-induced damage in human skin keratinocytes.Methods Cultured human keratinocyte HaCaT cells were divided into several groups to be treated with nicotinic acid,UVB irradiation,LY294002(an inhibitor of Akt),U0126(an inhibitor of extracellular signal-regulated kinase(ERK)1/2),SB203580(an inhibitor of P38)alone or in combination for different durations.Then,Western blot was performed to quantify the phosphorylation levels of the protein kinase B(Akt)/MAPK pathwayassociated proteins including Akt,P38,JNK and ERK1/2,MTT assay to evaluate the activity of HaCaT cells,enzyme-linked immunosorbent assay to determine the levels of endothelin-1(ET-1)and basic fibroblast growth factor(bFGF)in the culture supernatant of HaCaT cells,and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)to evaluate the apoptosis in HaCaT cells.Results As Western blot showed,phosphorylated Akt,P38,JNK and ERK1/2 were markedly activated within 60 minutes after pretreatment with nicotinic acid and irradiation with UVB(all P ＜ 0.01),and the activation was more significant for phosphorylated Akt,P38,and ERK1/2 within 2 hours(all P ＜ 0.01).Nicotinic acid effectively suppressed the UVB-induced cell death and apoptosis in HaCaT cells.The levels of supernatant ET-1 and bFGF were significantly decreased in HaCaT cells treated with the above 3 inhibitors followed by UVB irradiation than in those treated with the inhibitors alone(all P ＜ 0.05),and nicotinic acid pretreatment only reversed the decrease in supernatant bFGF in HaCaT cells treated with SB203580 followed by UVB irradiation.Conclusion The Akt signaling pathway may play a regulatory role in the protection by nicotinic acid against UVB-induced damage in HaCaT cells.

Objective To investigate the effect of nuclear translocation of E2p45 related factor 2 (Nrf2)on the biological activity of melanocytes.Methods Plasmid vectors containing wild-type nrf2 gene (pcDNA-nrf2) and nls-deleted nrf2 gene (pcDNA-nrf2△nls) were constructed.B10BR normal murine melanocytes were classified into three groups,i.e.,untransfected group,wild-type nrf2 group transfected with pcDNA-nrf2,and mutated nrf2 group transfected with pcDNA-nrf2△nls.Each of the above groups were further divided into three subgroups:control subgroup receiving no treatment,hydrogen peroxide (H2O2) subgroup treated with H2O2 of 200 μmol/L for 24 hours,and combined subgroup pretreated with tert-butyl hydroquinone (TBHQ) followed by treatment with H2O2 of 200 μmol/L for 24 hours.Subsequently,methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of cells,dopa oxidation assay to determine tyrosinase activity,Transwell assay to estimate cell migration ability,Western blot to quantify the expressions of Nrf2 and his tag fusion protein.Results TBHQ significantly enhanced the nuclear expression of Nrf2 in B10BR cells transfected with pcDNA-nrf2 or pcDNA-nrf2△nls (both P ＜ 0.01).No significant difference was observed in tyrosinase activity between untreated wild-type nrf2 group,mutated nrf2 group,and untransfected group (P ＞ 0.05).There was a statistical decrease in tyrosinase activity in the two H2O2-treated transfected groups compared with the untreated transfected groups (both P ＜ 0.05),and the decrease was reversed by TBHQ pretreatment in the wildtype nrf2 group (P ＜ 0.05),but not in the mutated nrf2 group (P ＞ 0.05).Further more,the proliferative activity of B10BR cells experienced no obvious changes in the wild-type nrf2 group (P ＞ 0.05),but was significantly reduced in the untransfected group (P ＜ 0.05) and mutated nrf2 group (P ＜ 0.01) after the H2O2 treatment compared with the corresponding untreated groups.TBHQ could protect the pcDNA-nrf2-transfected B10BR cells,but not pcDNA-nrf2△nls-transfected B10BR cells,from H2O2-induced oxidative damage.Transwell assay showed no significant difference in migration ability among these nine groups (P ＞ 0.05).Conclusions Abnormal nuclear translocation of Nrf2 could affect antioxidant activity of,proliferative activity of and tyrosinase activity in melanocytes.TBHQ may enhance the tyrosinase activity in,proliferative activity and antioxidant activity of melanocytes via activating the nuclear expression of wild type Nrf2.

Objective To explore the preventive effect of Fluconazole on invasive fungous infections in premature infants.Methods Two hundred and thirty-four cases of preterm infants hospitalized in Jiangmen Central Hospital from Feb.2008 to Oct.2013 were analyzed by retrospective study.The subjects were divided into 3 groups based on whether Fluconazole was used and it was used orally or intravenously for the prevention of invasive fungous infections in premature infants.The Fluconazole prophylaxis was not given to group A,whereas oral Fluconazole prophylaxis [6 mg/(kg · times),every other day] and intravenous Fluconazole prophylaxis [3 mg/ (kg · times),once every 3 days] were applied in group B and group C.The incidence of invasive fungous infections and the clinical effects of Fluconazole on the 3 groups were evaluated and compared.Results The number of cases analyzed in the study were 53,115 and 66 cases in group A,group B and group C,respectively.There were no significant differences between the 3 groups in terms of gender,gestational age,birth weights,length of hospitalization,intravenous nutrition,and number of peripherally inserted central catheter,antibiotics and invasive mechanical ventilation use days (all P ＞ 0.05).The incidences of invasive fungal infections were 11.32％ (6/53 cases),6.96％ (8/115 cases) and 0 in group A,group B and group C,respectively.The prophylaxis effects between the 3 groups were significantly different (x2 =7.078,P =0.029).Group A and group B were not significantly different (x2 =0.905,P =0.342),but group C was better than group A and group B,and the differences were significant (P =0.007,0.028).Conclusions Prevention of invasive fungous infections by intravenous Fluconazole has good curative effects,and which can be used as the first choice.