Objective To investigate the treatment of complex fractures of distal femur with open reduction and internal fixation with anatomical plate. Methods From January 2001 to June 2004, 67 cases of complex fractures of distal femur were treated with open reduction and anatomical plate internal fixation. The allogeneic bone was grafted for 23 cases, ilium was grafted for 35 cases, ilium and fibula were grafted for 9 cases. Bio- resorbable membranes were used between quadriceps femoris and femur. 13 cases were aided with patella traction. All the patients received fumigation and rinsing with Chinese herbs and CPM(continuous passive motion) after operation. Results All the 67 cases were followed up for 10 to 26 months. All the fractures got united. The postoperative knee functions were excellent in 36 cases, good in 23 cases, fair in 5 cases, and poor in 3 cases according to Kolment standards. The excellent and good rate was 88.1% . Conclusions Open reduction, internal fixation with anatomical plate, bone grafting, local use of bio- resorbable membranes, patella traction when necessary, fumigation and rinsing with Chinese herbs and CPM after operation are good measures to treat complex fractures of the distal femur, because they can provide stable fixation, allow early postoperative functional training, and prevent adhesion of quadriceps femoris and stiffness of the knee joint

Objective To investigate the clinical application value of hand-assistant laparoscopy and total laparoscopy in radical right colectomy.Methods Five hundred and twenty-four patients were selected as our subjects who performed by hand-assistant laparoscopic radical right colectomy (213 cases) or total laparoscopic radical right colectomy (293 cases) in colon cancer therapy.The operation durations,bleeding volumes,postoperative pain scores,postoperative draining volumes,recovery time of gastrointestinal function,hospitalization times,hospitalization expenses,postoperative complications and cosmetic satisfactory scores were recorded and compared.Results The operation durations,bleeding volumes,postoperative draining volumes in hand-assistant laparoscopic group were (112.4±52.9) min,(105.6 ± 49.5) ml,(28.6± 15.7) ml,lower than those in total laparoscopic group((148.5±61.7) min,(171.5±72.4) ml,(28.6±15.7) ml;P=0.032,0.041,0.039).The recovery time of gastrointestinal function of hand-assistant laparoscopic group were (69.4±21.8) h,longer than total laparoscopic group ((53.4 ± 21.7) h;P =0.008).The hospitalization times,hospitalization expenses,postoperative complications of hand-assistant laparoscopic group were as same as total laparoscopic group(P＞0.05).Hand-assisted laparoscopic group mild pain in 35 cases,121 cases of moderate and severe 75 cases;Total 96 cases of mild pain in laparoscopic group,146 cases of moderate and severe in 51 cases,the difference was statistically significant(P=0.027).Beauty score incision of laparoscopic group was higher than in the whole hand-assisted laparoscopic group,the difference was statistically significant ((4.3 ± 0.9) score vs.(2.4±0.6) score;P=0.002).Conclusion Hand-assistant laparoscopic radical right colectomy is convenient and more quickly for operation.Total laparoscopic radical right colectomy has advantage in terms of less injury and the patients are more satisfactory.For practiced operators,total laparoscopic radical right colectomy is recommended cosmetic satisfactory score.

OBJECTIVE:To establish the method for the detection of the known glucagon-like peptide 1 receptor (GLP1R) gene mutation site rs3765467(NT_007592.16,position:39065819),and to evaluate its accuracy and practicability. METHODS:Peripheral venous blood samples of 72 healthy subjects were collected in medical examination center of our hospital during Oct. 2015-Feb. 2016. The whole blood DNA was extracted by column extraction method. After amplified by touch down PCR,high reso-lution melting(HRM)method was adopted to analyze amplified product. Sequencing verification by double stranded chain termina-tion method(Sanger sequencing method)was performed for 38 test samples. The results of 2 methods were compared. RESULTS:The results of mutation scanning showed that there were 39065817 and 39065819 polymorphism sites in amplified segments. Four types of mutations were detected by HRM method [GCG/GCG,GCA/GCG or ACG/GCG,GCA/GCA or ACG/ACG,A(G) CA(G)],but 6 types of mutations was detected by Sanger sequencing method [GCG/GCG,ACG/GCG,ACG/ACG,A(G)CA (G),GCA/GCG,GCA/GCA]. CONCLUSIONS:HRM method can identify GCG/GCG and A(G)CA(G)genotype,but can not identify GCA/GCG and ACG/GCG heterozygous mutation,GCA/GCA and ACG/ACG homozygous mutation. The method is not suitable for the detection of single nucleotide polymorphism for multiple neighboring sites. In the detection of single nucleotide mu-tation,economical and simple method should be selected after comprehensive analysis of sequence.

The aims of this article is to provide the quality control requirements of preanalytical variation for the determination of trace element aluminum,arsenic,chromium,cadmium and mercury in samples of human origin,reduce the influence of preanalytical variation on the test results.Refer to the Clinical and Laboratory Standards Institute (CLSI) documents,Control of Preanalytical Variation in Trace Element Determinations and other references and guidelines,the methods of quality control of aluminum,arsenic,chromium,cadmium and mercury determination have been made,including:the factors needed to be considered in collection,preservation,transportation and other preanalytical factors,the abilities and considerations of laboratory staff,etc.Hope this article provide some useful suggestions and help to the laboratories of determination of aluminum,arsenic,chromium,cadmium and mercury in samples of human origin.

Objective To observe the inhibitory effect of andrographolide (AD) on the migration and invasion of human cervical cancer cells,and investigate the molecular mechanisms underlying the inhibitory effect of AD.Methods Human cervical cancer cell lines HeLa cells were cultured in vitro.After induced by 100 nmol/L of phorbol-12-myristate-13-acetate(PMA) for 24 h,different concentration of AD was co-incubated for another 24 h.Cell viability was detected by methyl thiazolyl tetrazoliym (MTT) assay.Chamber invasion assay was used to observe the migration and invasion,respectively.Expression of matrix metalloproteinase-9 (MMP-9) and translocation of nuclear factor kappa B (NF-κB) p65 subunit were detected by Western blot.Results The concentration (0 ～ 10 μmol/L) of AD exhibited a non-cytotoxic effect on HeLa cells.10 μmol/L of AD decreased PMA-induced migration by 49％ and invasion by 52％.Western blot showed that AD suppressed PMA-induced MMP-9 protein expression in a dose-dependent manner.Treatment of AD could decrease the translation of p65 in HeLa cells.Conclusions AD inhibits MMP-9 expression mediated by NF-κB,which lead to the suppression of migration and invasion of cervical cancer cells.

Objective To observe changes of cognitive function and the expression of tumor necrosis factor alpha(TNF-α),interleukin 10(IL-10) in hippocampus of diabetic rats,and assess the role of inflammation in the possible pathogenesis of diabetic encephalopathy (DE).Methods 30 male SD rats were randomly divided into control group and diabetes mellitus group.After 4 weeks of feeding high fat diet,diabetes mellitus group according to 30mg/kg injected with streptozotocin to establish type 2 diabetic rat model.At the end of the experiment,cognition were evaluated using water maze test.The concentration of beta-amyloid(Aβ) in hippocampus of diabetic rats were detected through enzyme linked immunosorbent assay,and the expression of TNF-α,IL-10 were detected by Western blotting.The expression of Aβ,TNF-α,IL-10 were observed through immunohistochemistry.Results Time spent in the target quadrant in diabetes mellitus group was shorter than that in control group ((38.21± 3.68)s vs (42.10±2.62)s,t=3.105,P＜0.01).The frequency of crossing original platform site was less than that in control group((2.62±0.77) vs(3.69±0.95),t=3.184,P＜0.01).Compared with control group the expression of Aβ,TNF-α were higher(BothP＜0.01),and IL-10 were lower(P＜0.01)in diabetes mellitus group.The positive expression of Aβ,TNF-α were obviously and IL-10 were less obviously observed in diabetes mellitus group according to immunohistochemistry.Conclusion The cognitive decline in diabetic rats is possibly related to inflammatory cytokines expressing out of balance.

A series of valproic acid salicylanilide esters were designed and synthesized based on the principle of prodrug. The structures of the target compounds were confirmed by MS, 1H NMR and 13C NMR. Anti-tumor activities of these compounds against K562, A549, A431 cells in vitro were investigated by MTT assay and SRB assay. The results indicated that the compounds 6h-6j were found to have stronger cell growth inhibitory action than gefitinib, and comparable to niclosamide, which are worth to be intensively studied further.

Objective To investigate the dates of 477 Chinese prenatal screening centers for previous half year analyse prena-tal screening status and provide recommendations for quality control.Methods All China prenatal screening centers were sent the data via the National Quality Assessment Scheme.This covered the software used,the risk cut-offs,monthly sample throughput,monthly median MoM of AFP,hCG,β-HCG,freeβ-HCG and uE3,monthly screening positive rate for trisomy 21,trisomy 18 and Open Neural Tube Defect.Results Screening protocol were versatile,with 73.48% (133/181)used two-marker model,24.31% (44/181)used three-marker model and 2.21% used four-marker model.About the software used, 350 laboratories never updated the screening parameters,89 laboratories had updated their median or parameter by manufac-turers,24 laboratories had updated the parameters by themselves.Cut-offs differed between laboratories.59.91% (275/459) used 1/270 as their cutoffs for trisomy 21.66.22% (296/447)used 1/350 as their cutoff for trisomy 18 and 96.52% (361/374)used cutoffs between 2.0~2.5 MOM for ONTD.Results of monthly median MOM:the percentage of laboratories that all six monthly median MOM within the target of 0.90~1.10 was 46.69% (155/332)for AFP,20.0% (4/20)for hCG, 29.17% (28/96)forβ-HCG,15.66% (31/198)for freeβ-HCG and 4.82% (11/228)for uE3.The percentage of laborato-ries that all six monthly median MOM within the target of 0.95~1.05 was 14.16% (47/332)for AFP,0% (0/20)for hCG,4.17% (4/96)forβ-HCG,12.63% (25/198)for freeβ-HCG and 4.82% (11/228)for uE3.About screening positive rate,there were difference of trisomy 2 1 positive rate in the same laboratory within the six month.Conclusion There were variations types of screening protocol,different kinds of soft platform,randomness of choice or update of medians or other important parameters and great difference in awareness of quality control.It would be taken effective and practical quality control measures to help laboratories improve quality control.

Objective To investigate the current situation in application of median levels in second trimester in prenatal screening laboratory throughout China and to understand the methods and importance of quality control.Methods Of those laboratories that participated in the national external quality assessment (EQA) for second trimester maternal serum screening in 2013, 442 submitted the medians for α-fetal protein (AFP), human chorionic gonadotropin (hCG), β-hCG, free β-hCG and unconjugated estriol (uE3) and related information.We categorized the medians according to the platform.There were 170, 210, 31, 26, 13, 11 and nine laboratories using the detection systems from Perkin Elmer, Beckman Coulter, Fenghua, Roche, Darui, Siemens and Abbott, respectively.The method of setting up the medians for each laboratory was analyzed.The number of laboratories, which was the outliers, was determined according to the reported medians on the third day of each week from 14th to 20th gestational weeks.Single sample t-test was applied to compare the result of this study with published results from a multi-center study of pregnant women in China.Results Among all laboratories, 19.0％ (84/442) of the recruited laboratories setup the median on their own.And 4.3％ (19/442) did this with their vendors;76.7％ (339/442) used the default median from the software, among them 97.9％ (332/339) did not verify their medians and 84.3％ (280/332) had no idea of how to do.Medians of AFP and uE3 increased with gestational age, while medians of hCG, β-hCG and free β-hCG decreased in all detection systems.More outliers were found for AFP, free β-hCG and uE3 medians in Perkin Elmer system, which scattered in each gestational age.A higher proportion of AFP in Fenghua system, free β-hCG in Darui system and uE3 in Siemens system were reported [9.5％ (20/210), 13.2％ (12/91) and 23.4％ (18/77), respectively].The medians of AFP and β-hCG using Perkin Elmer platform were compared between this study and the previous multi-center study in Chinese women.The results did not show significant difference in AFP median at 14th and 18th week of gestation (both P ＞ 0.05), while higher medians in AFP at 15th, 16th, 17th, 19th and 20th week of gestation (t=-7.257,-3.204,-5.479,-3.253 and-20.089, respectively, all P ＜ 0.05) and higher β-hCG median at 14th to 20th week of gestation (t=-80.749,-199.779,-142.601, -90.732,-72.984,-47.935 and-39.409, respectively, all P ＜ 0.01) were found in the previous study.Conclusion Most laboratories in our country never verify their medians for second trimester screening program and the medians vary among different settings.

Objective To explore the expressions of Toll-like receptors (TLRs) 2 and 4 in mouse skin during early immune responses against Sporothrix.Methods A total of 60 BALB/c mice were randomly and equally divided into an experimental group and a control group to be intracutaneously injected with Sporothrix conidium suspensions at a concentration of 1 × 106 cfu/ml and sodium chloride physiological solution respectively.Five mice were sacrificed before the injection,and at 6,12,24,48,and 96 hours after the injection in each group,blood samples were obtained from the mice before sacrifice,and skin tissue specimens were resected from the area around the injection sites after sacrifice.Realtime fluorescence-based quantitative PCR was performed to quantify the mRNA expressions of TLR2 and TLR4,and immunohistochemical staining to observe the protein expressions of TLR2 and TLR4 in mouse skin specimens.Enzymelinked immunosorbent assay (ELISA) was conducted to determine the levels of interleukin 12 (IL-12) and tumor necrosis factor α (TNFα) in serum samples from the mice.Results After injection of Sporothrix conidium suspensions,the mRNA expression level of TLR2 gradually increased and peaked at 24 hours,which was 18.8 times that in the control group at 6 hours and 34 times at 24 hours.In addition,the mRNA expression level of TLR4 in the experiment group reached a peak,and was 56.7 times that in the control group at 6 hours after injection,then gradually decreased and reached the nadir at 96 hours.As immunohistochemical staining revealed,TLR2 and TLR4 were apparently expressed in both keratinocytes and macrophages in skin specimens from the experimental group,but not obviously in those from the control group.No significant differences were observed between the experimental group and control group in serum levels of IL-12 or TNF-α at any of the sampling time points.Conclusion TLR2 and TLR4 may play a favoring role in immunological defense by participating in the recognition of Sporothrix by keratinocytes and macrophages in mouse skin.