0.05) The sensitvity is over 5 parasites/108 RBC. However, it is not satisfactory by using this method to detect P.vivax antigen.

This paper reports an improved McAb-ELISA test in detecting blood stage P.vivax antigen in which the plates were coated with rabbit anti-P.cynomolgi poly-antibody and two monoclonal antibodies were used together for reaction. The coincidence rate with microscopically conformed P.vivax blood samples was 94.3% and the coincidence rate with microscopically negative blood samples was 96.1%. The sensitivity was over 1 parasite/106 erythrocytes.

OBJECTIVE:To establish a method for simultaneous determination of sodium valproate(VPA)and its metabo-lite 2-propyl-2-pentenoic acid (2-ene-VPA) in human plasma. METHODS:Plasma sample was extracted with cyclohexane and experienced derivatization with 2,4′-dibromoacetophenone using n-octanoic acid as an internal standard. RP-HPLC method was adopted. The determination was performed on Zorbax SB-C18 column with mobile phase consisted of acetonitrile-water(65∶35,V/V)at the flow rate of 1 mL/min. The column temperature was set at 35 ℃,and UV dectection wavelenth was set at 258 nm. The sample size was 20 μL. RESULTS:The linear range of VPA and 2-ene-VPA were 5.0-200.0,0.5-20.0 μg/mL(r=0.999 9, n=5). The limits of quantification were 5.0,0.5 μg/mL. RSDs of inter-day and intra-day were all lower than 5%. Method recov-eries were 95.99%-98.80%and 97.40%-98.17%,and extraction recoveries were 80.46%-86.23%and 80.45%-85.61%. The plas-ma concentrations of VPA in 10 epileptic children were 27.4-93.2 μg/mL,and those of 2-ene-VPA were 0.85-3.94 μg/mL,respec-tively. CONCLUSIONS:The method is simple,specific and suitable for plasma concentration determination and pharmacokinet-ic study of VPA.

BACKGROUND: Acetabular fracture is considered to be one of the most challenging fractures because of its complex anatomical structure, which makes it more difficult to treat. At present, open reduction and internal fixation are still the standard treatment methods for displaced acetabular fractures. The patient-specific pre-contour reconstruction template made by three-dimensional (3D) printing technology combined with preoperative virtual surgery plan can reduce the invasiveness of surgery and simplify the operation process. OBJECTIVE: To evaluate the effect of 3D printing combined with virtual surgical planning in the treatment of acetabular fractures compared with traditional reduction and reconstruction of acetabular fractures. METHODS: Totally 25 patients were selected from Affiliated Hospital of Chengde Medical College from October 1, 2017 to March 1, 2018, including 14 males and 11 females, at the age of 21-60 years old. They were divided into experimental group (n=12) and control group (n=13) by computer random grouping method. In the experimental group, printing technology combined with virtual pre-contour reconstruction plate fixation was performed. In the control group, intraoperative contour reconstruction plate fixation was conducted after reduction. Postoperative X-ray and non-contrast CT scan was used to analyze fracture reduction in two groups. After the operation, the patients in the two groups were followed up for visual analogue scale score, Majeed function score and complications. This study was approved by the Ethics Committee of Affiliated Hospital of Chengde Medical College (approval No. LL007). RESULTS AND CONCLUSION: (1) X-ray films showed that the reduction effect of the experimental group was better than that of the control group (P=0.038). Non-contrast CT images showed that the reduction displacement difference of the experimental group was better than that of the control group before and after operation [(12.43±7.58) mm, (9.408±8.27) mm, P < 0.05]. (2) At 6-12 months after surgery, visual analogue scale scores were lower in the experimental group than in the control group [(1,6±0.6), (3.3±1.3), P < 0.05], There were no significant differences in Majeed function scores between the two groups in the final follow-up (P=0.079). (3) Complications, such as delayed healing, failure of internal fixation or deep vein thrombosis, were not found; and no biocompatibility adverse reactions related to the implanted device occurred in the two groups after surgery. (4) Results suggested that 3D printing technology assisted virtual operation plan can improve the reduction quality in the fixation of acetabular fracture, improve the operative effect of acetabular fracture, and reduce the postoperative pain of patients.

ObjectiveTo investigate the effects of maternal rat with marginal iodine deficiency on the nerve and intelligence development in its offspring and the related mechanisms.MethodsThe rat model of marginal iodine deficiency was established on the basis of epidemiological characteristics and iodine metabolism characteristics of rats reported in literatures.Marginal iodine deficiency rats were given 3 μg iodine a day ; normal control rats were given 4 μg iodine daily ; iodine deficiency rats were given 1.2 μg iodine daily,respectively.Western blot was used to detect the protein levels of brain derived neurophic factor (BDNF) and early growth response factor 1 ( EGR1 ) in the hippocampus of 7 days newborn rats.Immunohistochemistry was used to measure c-jun and c-fos expressions in the hippocampal CA1 region of 40 days rats.Water maze method was used to measure the ability of learning and spatial memory.Results FT4 level in rats of marginal iodine deficiency group during pregnancy decreased by about 30％.Seven days after birth,the contents of EGR1 and BDNF proteins in the hippocampus of the offspring from marginal iodine deficiency group were significantly decreased compared with normal control group (both P ＜ 0.05 ).Forty-day-old newborn rats in marginal iodine deficiency group also showed significantly decreased c-jun and c-fos expressions in the hippocampal CA1 region ( both P＜0.05 ),along with a decreased trend of spatial learning and memory ability.Conclusions FT4 level will significantly decrease after pregnancy if rats have marginal iodine deficiency,affecting the expression of related proteins in the brain of newborn rats and having a negative impact on offspring brain development.

Objective To investigate the clinical manifestations,laboratory findings and genetic characteristics of two cases with simple virilizing (SV)21-hydroxylase(21OHD). Methods Clinical manifestations and laboratory data were obtained. The promoter and coding areas of CYP21A2 gene were directly sequenced. In silico analysis were used to predict the function of mutations. Results Two patients presented severe virilism. The laboratory examinations showed that plasma ACTH,aldosterone,androstendione and testosterone were significantly increased compared with normal individuals. VCT scan showed hyperplasia of bilateral adrenal. Direct sequencing of CYP21A2 gene showed two complex mutations-H62L/V69L and I2g/10InsL,which were not reported previously. In silico analysis(Polyphen)showed the novel mutation V69L could probably damage the function of CYP21A2 protein. Conclusion The combined mutations,H62L/V69Land I2g/10InsL,could be associated with SV type 21-OHD phenotype.

Objective:To study whether Noxa mediates cell death induced by etoposide in the human neuroblastoma(NB)cells.Methods:NB cells(TB3 and TB8) were treated with different concentrations of etoposide(0, 0.125, 0.25, 0.5 0.75, 1.0 mg/L), and the cell survival was detected by CCK8 assay.After treated with etoposide, NB cells were collected at different time points, then total RNAs were isolated and RT-qPCR was performed to detect the mRNA expression of Noxa.At the same time, the whole cell lysates were extracted and western blot was performed to detect the protein expression of Noxa.In order to evaluated the effect of Noxa on etoposide-induced cell survival, Noxa siRNA was transfected into NB cells, then CCK8 assay was performed.Results:After treatment with different concentrations of etoposide(0.125 mg/L、0.25 mg/L、0.5 mg/L、0.75 mg/L、1.0 mg/L ), the survival rates of TB3 cells were(73.13±8.45)%, (56.18±10.50)%, (33.90±4.17)%, (26.76±6.67)%, (13.49±0.58)%, respectively(compared with the control group, P<0.01); the survival rates of TB8 cells were(71.06±6.96)%, (37.45±0.68)%, (25.53±3.70)%, (20.28±2.75)%, (10.09±2.52)%, respectively(compared with the control group, P<0.01).The mRNA and protein expression of Noxa in NB cells were both increased in a time-dependent manner after treated with etoposide.The siRNA of Noxa could reduce the expression of Noxa in TB3 and TB8 cells after transfection.Treated with etoposide 0.5 mg/L, cell survival rates of TB3 cells tranfected with control siRNA, Noxa siRNA1, Noxa siRNA2 were(45.12±13.58)%, (72.70±21.34)%, (52.08±20.36)%, respectively; cell survival rates of TB8 were(35.52±0.38)%, (63.94±0.10)%, (50.27±1.62)%, respectively(compared with the control group, P<0.01); Treated with etoposide 1.0 mg/L, cell survival rates of TB3 cells tranfected with control siRNA, Noxa siRNA1, Noxa siRNA2 were(13.26±1.84)%, (51.08±2.41)%, (42.80±1.42)%, respectively(compared with the control group, P<0.05); cell survival rates of TB8 were(22.22±3.39)%, (58.00±11.37)%, (40.55±6.94)%, respectively(compared with the control group, P<0.05). Conclusion:Pro-apoptotic molecular Noxa mediated the Etoposide-induced cell death in NB cells(TB3 and TB8) in time and concentration-dependent manner.

BACKGROUND:Cold trypsin digestion is rarely reported to culture umbilical cord mesenchymal stem cells. OBJECTIVE:To compare the biological characteristics of umbilical cord mesenchymal stem cells cultured by cold trypsin digestion and tissue explant method. METHODS:Human umbilical cord mesenchymal stem cells were isolated, purified and passaged using cold trypsin digestion and tissue explant method, and then the first adhesion time and cellcycle were recorded. Morphology of umbilical cord mesenchymal stem cells was observed under inverted phase contrast microscope to draw growth curve of cells at passage 3. Flow cytometry was used to detect the surface markers of passage 3 umbilical cord mesenchymal stem cells, and Nestin expression was detected in passage 3 cells after 3 days culture in neural induction medium by fluorescence immunochemistry staining. RESULTS AND CONCLUSION:These two methods were both successful to harvest umbilical cord mesenchymal stem cells, but the first adhesion time was earlier in cells cultured by cold trypsin digestion than tissue explant method (P<0.05), and there was no difference in primary cellcycle (P>0.05). Under the inverted microscope, cells grew adherently and presented fibroblast-like shape. However, the minority of primary cells under induction of cold trypsin digestion was polygonal, irregular, and had larger cellvolume than those cultured by tissue explant method. Passage 3 cells cultured by tissue explant method showed faster proliferation rate than those cultured by cold trypsin digestion, and at logarithmic growth phase, cells cultured by these two methods were significant different in cellnumber (P<0.05). Two types of cells had a uniform cellphenotype, both of which expressed CD29, CD105, but did not express CD34, CD45. Under induction, passage 3 cells cultured by these two methods were both positive for nestin. These findings indicate that these two methods can both be used to culture umbilical cord mesenchymal stem cells, but the tissue explant method is more suitable for culture of umbilical cord mesenchymal stem cells and exhibits less damage to cells.

Objective To investigate the mechanism of venous reverse-flow flap in the differentperiod after operation.Methods The rabbits wero randomly allocated into 3 groups.In group A,including saphenous artery and venae commutante.In group B,saphenous artery without venae commutante.In group C,surface seeping and saphenous artery and venae commutante.Flap appearance,intravenous pressure,vessel diameter,mierocircular and histological examination were mea8ured.Results The difference of introvenous pressure between group A.B and C was obvious.Reverse flow WaS found in group A and C group through microcirculation observation 2 hours post-operation.Venous valve lose efficacy while the vessel diameter wes at maximum just after the pressure peak.Conclusion Venous retrograde return in reverse-flow island flaps can be achieved more easily through"incompetent valves route"than through "communicating and collaterall by pass route".By pass route is a supplementary way.Surface seeping Can slighfly relieve the venous pressure but can cause infection.