Some published research reports on phthalate esters (PAEs) in the municipal sewage sludge were reviewed in the present paper, and aimed at the problems in the field, some preventive measures and the related research emphases were put forwarded also.

Objective To improve and establish an RP-HPLC method for the determination of related substances of piracetam and its injection. Methods Using thermo syncronis C18(4.6 mm ×250 mm,5μm)chromatographic column, mobile phase was acetonitrile-phosphate buffer (adjust pH of 1.0 g/L potassium hydrogen phosphate solution to 6.0 with phosphoric acid), gradient elution with initial mobile phase 5% acetonitrile.The flow rate was 1 mL/min, detected at 205 nm.Five calibration factors of the known impurities were separately measured.the related substances were determined using main component self-compare with calibration factor.Results Piracetam and their related substances were well separated.The calibration curves of five known impurity A~E were linear within the concentration range measured.The average recoveries were 99.46%,98.21%,97.22%,100.23% and 97.58%, respectively(n=9), RSDs of the average recoveries were lower than 2.0%.The calibration factors of five known impurity A ~E were 1.1, 1.4,1.5,0.8, 1.4, respectively.Conclusion The method can be used to determine the related substances in piracetam and its injection.The related substances can be determined accurately using main component self-compare with calibration factor.

Objective To study the degradation of hepatitis C virus(HCV) genome before and after methylene blue photochemistry(MB-P) treatment of the plasma.Methods MB was added to HCV positive plasma to a final concentration of 1.0?mol/L.The plasma was then exposed to 30000 Lux fluorescence.Plasma samples were then collected at different times of exposure.Real-time PCR was used to quantitatively study the course of the HCV-RNA degradation.The whole genome was screened for integrity by RT-PCR with 8 pairs of specific primers which targeted sequential overlapping sections of HCV genome.Results Results of real-time PCR showed that the copy number of HCV-RNA was continuously decreasing during MB-P treatment.RT-PCR results showed that the reactivity of different sections of the HCV genome to MB-P was significantly different and indicated that the 5' end and 3' end were more stable. Conclusion MB-P treatment could degrade HCV RNA with various sensitivities to different sections of the genome.RNA degradation may play an important role in plasma virus inactivation.Detection of HCV genomic RNA might be clinically useful to monitor the process and efficiency of virus inactivation.

Objective To analyze the relationship between dynamic change of GPI-80 and disease severity and prognosis of childhood anaphylactoid purpura. Methods Patients were collected and divided into three groups according to their clinical features: purpura group (purpura only), mixed group (purpura + arthritis + gastrointestinal bleeding) and nephritis group. There were 20 patients in each group. GPI-80 expression on the neutrophils was detected by flow cytometry during acute and regressive phases of the disease. GPI-80 expression was compared among different groups and different phases. Renal biopsies were performed in 20 nephritis patients. Results GPI-80 expression was significantly increased in all patient groups compared with that in the normal control (P 0.05). No significant difference of GPI-80 expression was found among 20 nephritis patients with different pathological patterns. Forty-two patients (10 in purpura group, 15 in mixed group, and 17 in nephritis groups) were followed up and GPI-80 expression was detected at the time of discharge and 2 weeks after discharge, the results showed that GPI-80 expression was decreased from 93.26% (?7.89%) at acute phase to 91.37% (?6.9%) at regressive phase with an average interval of 13.5 days. Most of them (35/42) further decreased to 38.44% (?7.8%) in 2 weeks after discharge. GPI-80 expression remained high in 7 patients for 2 weeks after discharge and relapsed in 5 patients within 1 month after discharge. Conclusions High GPI-80 expression is related to the severity of the disease. The decrease of GPI-80 takes place later than the improvement of clinical symptoms. Children with persistently high GPI-80 expression are likely to relapse. It seems that there is no correlation between GPI-80 expression and different pathological patterns of nephritis.

Objective To realize the contamination status of dental unit waterlines (DUWL)in general hospitals, and provide scientific evidence for making preventive measures.Methods Three hospitals were selected for study, water source adopted by hospital A,B and C was running water,reservoir water,and filtered water through reverse osmosis filtration system respectively,specimens of dental handpiece spray water and flushing water of dental chair units were collected quarterly,total bacterial colony in water were detected.Results The qualified rate of source wa-ter,handpiece spray water,and flushing water in hospital A was 75.00%(3/4),0 (0/40)and 0 (0/40)respectively,col-ony count of handpiece spray water and flushing water was (1.20×103 -5.53×104 )CFU/mL(M=3.80×104 CFU/mL) and (2.11×104 -1.66×105 )CFU/mL(M=4.80×104 CFU/mL)respectively.The qualified rate of source water,hand-piece spray water,and flushing water in hospital B was 50.00%(2/4),60.00%(24/40)and 72.50%(29/40)respectively, colony count of handpiece spray water and flushing water was (0.00 -3.71 ×106 )CFU/mL(M=83.00 CFU/mL)and (0.00-2.39×106 )CFU/mL(M=72.00 CFU/mL)respectively.The qualified rate of source water,handpiece spray wa-ter,and flushing water in hospital C was 100.00%(4/4),55.00%(22/40)and 65.00%(26/40)respectively,colony count of handpiece spray water and flushing water was (0.00-6.20×103 )CFU/mL(M=96.00 CFU/mL)and(0.00-1.63×103 )CFU/mL(M=87.50 CFU/mL)respectively.Conclusion Water of DUWL in general hospitals is seriously con-taminated,disinfection and standardized management of source water and DUWL must be strengthened.

Objective To analyze the clinical characteristics and treatment strategy of ankle supination fracture in children.Methods From January 2012 through July 2014,89 children were treated at our department for ankle fracture caused by supination sprain according to their medical history,physical examination,X-ray films and CT three-dimensional reconstruction of the ankle.Appropriate protocols were applied on the basis of Lauge-Hansen classification,type and displacement of their fractures.Of them,52 belonged to the supination-adduction type (the extramalleolus fracture was of Odgen type Ⅶ in 51 children whose epiphyseal plate of distal fibula had not been closed),35 to the supination-extorsion type (32 cases had tri-plane fracture and 3 Tillaux fracture),and the remaining 2 did not fit the Lauge-Hansen classification.Surgical treatment was applied in 32 cases and conservative treatment in 57 ones.Results All the children received outpatient follow-up from 12 to 24 months(mean,18 months).No bone nonunion,osteoarthritis,or fracture malunion was found.The American Orthopaedic Foot & Ankle Society scores averaged 92 points (range,from 88 to 100 points) at the last follow-ups.Conclusions In children whose epiphyseal plate is nearly closed,supination-adduction sprain likely causes an extramalleolus fracture of Salter-Harris type Ⅰ or type Ⅱ,but in children whose epiphyseal plate is unclosed,an epiphyseal fracture of Odgen type Ⅶ is inclined to happen.A Tillaux fracture or tri-plane fracture at the level of distal tibiofibular syndesmosis results often from supination-extorsion sprain in children.For fractures involving epiphysis or epiphyseal plate,anatomical reduction and proper fixation are critical to functional recovery and reducing complications.

Objective To investigate whether ultrasonic coupling agent (UCA)can produce shielding or antago-nistic effect on iodine disinfectant for preoperative skin disinfection.Methods Shielding or antagonistic effect of UCA on iodine disinfectant were detected by laboratory carrier immersion killing test and on-the-spot skin disinfec-tion test.Results Antagonistic effect:after the mixing of iodophor with UCA,the average killing rate of iodophor containing available iodine 2 500mg/L and 625 mg/L to Staphylococcus aureus decreased from 100.00% to 99.67%-99.78% and 96.85 % - 98.25 %,respectively;the average killing rate to Escherichia coli decreased from 100.00% to 99.71 %-99.82% and 95 .93 %-98.56%,respectively.Shielding effect:after smearing with UCA, the average killing rate of iodophor and iodine tincture + alcohol to Escherichia coli decreased from 100.00% to 30.76% and 100.00% to 94.48%,respectively;the average killing rate to Staphylococcus aureus decreased from 99.99% to 55 .55 % and 100.00% to 98.22%,respectively.On-the-spot skin disinfection test:the killing rate of io-dophor and iodine tincture +alcohol to natural bacteria on skin surface were both 99.99%,after skin was smeared with UCA,the killing rate decreased to 92.62% and 93 .57%,respectively.Conclusion UCA remained on the oper-ative field has shielding and antagonistic effect on iodine disinfectant.

Objective:To express fusion protein of mouse thymic stromal lymphopoietin (TSLP) and HIV-1 gp120BAL V1/V2 subdomain in 293F cell.Methods:Full length of the V1V2 sequence from BAL isolate was fused with the C-terminus of mouse thymic stromal lymphopoietin (TSLP) and sub-cloned into pCEP-Pu vector to construct the eukaryotic expression plasmid-pCTV1V2BAL.The recombinant plasmid was confirmed by enzyme digestion and sequencing , then transfected into 293 F cells using PEI as a transfection reagent .The fusion protein was purified by metal chelate affinity chromatography and characterized by SDS -PAGE and Western blot . The epitopes of V1/V2 in fusion protein were identified by ELISA .Results:The SDS-PAGE and Western blot results showed that there were highly heterogeneous glycoprotein bands at the site between 35 kD and 55 kD, which reacted with anti-mTSLP rabbit polyclonal antibody and anti-His tag mouse monoclonal antibody .The ELISA analysis showed that antibodies to V 1/V2BAL existed in the sera of HIV-1 positive patients.Conclusion:The mTSLP-V1/V2 fusion protein was successfully expressed in 293F cells, which may be useful for HIV-1 vaccine research .

Objective: To explore the expression of Dickkopf-3(DKK-3) mRNA in oral squamous cell carcinoma(OSCC) and the relationship between the promoter methylation status and the carcinogenesis of OSCC. Methods: The expression of DKK-3 gene in 51 cases of OSCC and corresponding normal mucosa tissue was detected by PT-PCR and methylation-specific PCR, respectively. The relationship between DKK-3 and the clinical pathological features of the patients was analyzed with SPSS 13. 0. Results: The expression level of DKK-3 in OSCC group was lower than that in the control(t =-12. 580, P＜ 0. 05). The methylation rate of DKK-3 gene promoter region in OSCC group was significantly higher than that in the control(χ2 = 19. 273, P＜ 0. 05). The mRNA expression level of DKK-3 gene in OSCC with methylation group was lower than that in the control(t =-2. 817, P＜ 0. 05). Conclusion: Down-regulation of DKK-3 gene expression and hypermethylation of promoter region are important mechanisms in the pathogenesis of OSCC. The hypermethylation of DKK-3 promoter may be the main cause of transcriptional silencing.

Objective: To investigate the effect and related mechanism of apolipoprotein A5 (ApoA5) on adipogenic differentiation of human adipose-derived mesenchymal stem cells (AMSC). Methods: Subcutaneous adipose tissue was obtained from 40 patients undergoing abdominal surgery at our hospital from February to July 2015. After induction of human AMSC by collagenase digestion, the adipose tissue was induced to differentiate into mature adipocytes and treated with ApoA5 at 600 and 1 200 ng/ml, respectively (ApoA5 intervention groups). Cells treated without ApoA5 protein were used as control group. The cells were harvested on the 7th and 14th day of differentiation, and the following assays were performed: (1) the effect of ApoA5 on TG content was measured by a TG assay kit; (2) RT-qPCR assay was used to detect the effect of ApoA5 on aP2 and FAS mRNA expression; (3) the effect of ApoA5 on the expression of CIDEC mRNA and protein was detected by RT-qPCR and Western blot; (4) the effect of ApoA5 on the expression of C/EBPβ mRNA and protein was detected by RT-qPCR and Western blot; (5) using lentiviral transfection technique, we overexpressed the gene of CIDEC in AMSC and cells were divided into lentiviral negative control group, lentiviral over-expressed CIDEC group and lentiviral over-expressed CIDEC+ApoA5 intervention group (the ApoA5 intervention concentration was 1 200 ng/ml). Thereby, we examined the effect of ApoA5 on the above indicators in adipogenic differentiation of AMSC in the case of CIDEC overexpression. Results: (1) Effect of ApoA5 on TG content in AMSC: on the 7th and 14th day after the intervention, the TG levels were lower in ApoA5 600 and 1 200 ng/ml group AMSC than those in the control group (all P<0.05). (2) The effect of ApoA5 on the expression of aP2 and FAS mRNA in AMSC: on the 7th day after intervention, the expression levels of aP2 and FAS mRNA were significantly lower in ApoA5 600 and 1 200 ng/ml group than those in the control group (all P<0.05). On the 14th day after intervention, the expression levels of aP2 and FAS mRNA were lower in ApoA5 600 and 1 200 ng/ml group than those in the control group (all P<0.05). (3) The effect of ApoA5 on the mRNA and protein expression of CIDEC in AMSC: on the 7th day after intervention, the mRNA and relative protein expression levels of CIDEC were significantly lower in AMSC of ApoA5 600 and 1 200 ng/ml group than those of the control group (all P<0.05). On the 14th day after intervention, the mRNA and relative protein levels of CIDEC were further reduced in ApoA5 600 and 1 200 ng/ml AMSC groups than those in the control group (all P<0.05). (4) The effect of ApoA5 on C/EBPβ mRNA and protein expression in AMSC: on the 7th day after intervention, C/EBPβ mRNA and relative protein expression levels were significantly lower in ApoA5 600 and 1 200 ng/ml group than those in the control group (all P<0.05). On the 14th day after intervention, the levels of C/EBPβ mRNA and relative protein were lower in ApoA5 600 and 1 200 ng/ml group than those in the control group (all P<0.05). (5) The effect of ApoA5 on the content of TG in AMSC after CIDEC overexpression: on the 7th and 14th day after intervention, the TG contents in AMSC were higher in the lentivirus over-expressed CIDEC group than in the lentivirus negative control group (both P<0.05). However, TG contents in AMSC were similar between the over-expressed CIDEC group and the CIDEC+ApoA5 over-expression group (both P>0.05). (6) The effect of ApoA5 on the expression of aP2 and FAS mRNA in AMSC after CIDEC overexpression: on the 7th day after intervention, the expression levels of aP2 and FAS mRNA in AMSC were higher in the lentivirus over-expressed CIDEC group than in the lentivirus negative control group (both P<0.05). On the 14th day after intervention, the expression level of aP2 mRNA in the AMSC was higher in the lentivirus over-expressed CIDEC group than in the lentivirus negative control group (P<0.05). On the 7th and 14th day after intervention, the expression levels of aP2 and FAS mRNA in AMSC were similar between the lentivirus over-expressed CIDEC group and the lentivirus over-expressed CIDEC+ApoA5 group (all P>0.05). (7) The effect of ApoA5 on the expression of C/EBPβ mRNA and protein in AMSC after CIDEC overexpression: on the 7th day after intervention, the mRNA and relative protein expressions of C/EBPβ in AMSC were higher in lentivirus-overexpressed CIDEC group than in lentivirus negative control group (both P <0.05). On the 14th day after intervention, C/EBPβ mRNA and protein expression levels in AMSC were higher in the lentivirus over-expressed CIDEC group than in the lentivirus negative control group (both P<0.05). On the 7th and 14th day after intervention, the expressions of C/EBPβ mRNA and protein in AMSC were similar between lentivirus over-expressed CIDEC group and lentivirus over-expression CIDEC+ApoA5 intervention group (all P>0.05). Conclusions ApoA5 can inhibit the adipogenic differentiation of AMSC,and this effect may be mediated by down-regulating the expression of CIDEC. Furthermore, our results indicate that CIDEC could be considered as a key factor in adipogenic differentiation.