Application of omics in the pathogenesis and prediction of hypertensive disorders of pregnancy: a review / 预防医学

Abstract@#Hypertensive disorders of pregnancy (HDP) are a common severe complication during pregnancy, which is characterized by complex etiology, unclear pathogenesis and lack of effective tools for early diagnosis and prediction. Recently, the development of omics technology provides new insights into the research into HDP. Based on national and international publications from 2011 to 2022, this review summarizes the application of genomics, proteomics, metabolomics, and microbiomics in the pathogenesis and prediction of HDP, so as to provide insights into the prediction, prevention and precise treatment of HDP.

Interaction between Abnormal Prepregnancy Body Mass Index and Hyperlipidemia during Pregnancy on the Risk of Gestational Diabetes Mellitus / 中国医学科学院学报

Objective To explore the interaction between abnormal prepregnancy body mass index(pBMI)and high blood lipid level during pregnancy on the risk of gestational diabetes mellitus(GDM). Methods A total of 235 patients with GDM and no blood lipid-related diseases before pregnancy were selected from Hangzhou Women's Hospital during March 2017 to July 2018 as the GDM group.At a ratio of 1∶3,a total of 705 individual age-matched pregnant women with normal glucose metabolism during prenatal examination from the same hospital were selected as the control group.The generalized multifactor dimension reduction(GMDR)method was employed to characterize the possible interaction between pBMI-blood lipid and GDM.The cross-validation consistency,equilibrium test accuracy,and P value were calculated to evaluate the interaction of each model. Results GMDR model analysis showed that the second-order model including pBMI and gestational blood lipid level had the best performance(P=0.001),with the cross-validation consistency of 10/10 and the equilibrium test accuracy of 64.48%,suggesting that there was a potential interaction between pBMI and gestational high blood lipid level.After adjustment of confounding factors,the model demonstrated that overweight/obesity patients with high triglyceride(TG) level had the highest risk of developing GDM(OR=14.349,95%CI=6.449-31.924,P<0.001).Stratified analysis showed that overweight/obesity patients under high TG level group had a higher risk of developing GDM than normal weight individuals(OR=2.243,95%CI=1.173-4.290,P=0.015). Conclusions Abnormal pBMI and high blood lipid level during pregnancy are the risk factors of GDM and have an interaction between each other.Overweight/obese pregnant women with high TG levels are more likely to develop GDM.

Behavioral Responses of Pregnant Women to the Early Stage of COVID-19 Pandemic in the Network Era in China: Online Questionnaire Study

Purpose@#The aim of this study was to examine the behavioral responses of pregnant women during the early stage of Coronavirus Disease 2019 (COVID-19) outbreak. @*Methods@#We recruited 1,099 women to complete an online questionnaire survey from February 10 to February 25, 2020. The subjects were divided into two groups (the pregnant women group and the control group). @*Results@#Concerns about infection: most of the participants watched the COVID-19 news at least once a day. Protective behaviors: the utilization rate of pregnant women (often using various measures) was higher than that of nonpregnant women. Exercise: 30.6% of the pregnant women continued to exercise at home, whereas in the control group, this percentage was 8.4%. Spouse relationship: 38.8% of the subjects’ relationship improved, whereas only 2.3% thought the relationship was getting worse. @*Conclusion@#Pregnant women had some unique behavioral responses different from that of nonpregnant women. It is important to understand the behavioral responses of pregnant women in this network era.

HSV-1 H129-Derived Anterograde Neural Circuit Tracers: Improvements, Production, and Applications / 神经科学通报·英文版

Anterograde viral tracers are powerful and essential tools for dissecting the output targets of a brain region of interest. They have been developed from herpes simplex virus 1 (HSV-1) strain H129 (H129), and have been successfully applied to map diverse neural circuits. Initially, the anterograde polysynaptic tracer H129-G4 was used by many groups. We then developed the first monosynaptic tracer, H129-dTK-tdT, which was highly successful, yet improvements are needed. Now, by inserting another tdTomato expression cassette into the H129-dTK-tdT genome, we have created H129-dTK-T2, an updated version of H129-dTK-tdT that has improved labeling intensity. To help scientists produce and apply our H129-derived viral tracers, here we provide the protocol describing our detailed and standardized procedures. Commonly-encountered technical problems and their solutions are also discussed in detail. Broadly, the dissemination of this protocol will greatly support scientists to apply these viral tracers on a large scale.

Behavioral Responses of Pregnant Women to the Early Stage of COVID-19 Pandemic in the Network Era in China: Online Questionnaire Study

Purpose@#The aim of this study was to examine the behavioral responses of pregnant women during the early stage of Coronavirus Disease 2019 (COVID-19) outbreak. @*Methods@#We recruited 1,099 women to complete an online questionnaire survey from February 10 to February 25, 2020. The subjects were divided into two groups (the pregnant women group and the control group). @*Results@#Concerns about infection: most of the participants watched the COVID-19 news at least once a day. Protective behaviors: the utilization rate of pregnant women (often using various measures) was higher than that of nonpregnant women. Exercise: 30.6% of the pregnant women continued to exercise at home, whereas in the control group, this percentage was 8.4%. Spouse relationship: 38.8% of the subjects’ relationship improved, whereas only 2.3% thought the relationship was getting worse. @*Conclusion@#Pregnant women had some unique behavioral responses different from that of nonpregnant women. It is important to understand the behavioral responses of pregnant women in this network era.

Interfering effect of maternal cell contamination on invasive prenatal chromosome microarray analysis / 中华检验医学杂志

Objective@#To access the effect of maternal cell contamination (MCC) on the detection of copy number variation (CNV) by chromosome microarray analysis (CMA) in prenatal diagnostic samples.@*Methods@#Amniotic fluid DNA samples were collected from Department of Prenatal Diagnosis (Screening) Centre of Hangzhou Women′s Hospital from December 2016 to August 2018. Copy number variations (CNVs) were identified in these DNA samples by CMA and normal female genomic DNA was added to simulate different proportions of maternal cells contamination. The simulated samples were tested using an Agilent microarray chromosome chip 180K CGH (Agilent 180K CGH) and the results were analyzed by Agilent CytoGenomics software.@*Results@#The results showed that duplications of CNV could not be detected at>38.4% MCC. Deletions of CNV could not be detected at>41.3% of MCC. With the increase of MCC ratio, the detection rate of CNV decreased gradually. The array and software allowed detection of longer copy-number variations at higher levels of maternal cell contamination than shorter copy-number variations. For the same size CNV, the detection ability of the deleted CNV was slightly higher than that of duplication. In the male sample, the observable shift of the X/Y chromosome ratio at>10% MCC can be detected by the microarray.@*Conclusions@#When the MCC is higher than a particular ratio, it can affect the CMA detection of CNV. Based on the Agilent 180K CGH detection results for MCC mimics and the CNV detection specificity principle, the MCC threshold of the Agilent 180K CGH is set to 30% in view of conservative principles.

Interfering effect of maternal cell contamination on invasive prenatal chromosome microarray analysis / 中华检验医学杂志

Objective:To access the effect of maternal cell contamination (MCC) on the detection of copy number variation (CNV) by chromosome microarray analysis (CMA) in prenatal diagnostic samples.Methods:Amniotic fluid DNA samples were collected from Department of Prenatal Diagnosis (Screening) Centre of Hangzhou Women′s Hospital from December 2016 to August 2018. Copy number variations (CNVs) were identified in these DNA samples by CMA and normal female genomic DNA was added to simulate different proportions of maternal cells contamination. The simulated samples were tested using an Agilent microarray chromosome chip 180K CGH (Agilent 180K CGH) and the results were analyzed by Agilent CytoGenomics software.Results:The results showed that duplications of CNV could not be detected at >38.4% MCC. Deletions of CNV could not be detected at >41.3% of MCC. With the increase of MCC ratio, the detection rate of CNV decreased gradually. The array and software allowed detection of longer copy-number variations at higher levels of maternal cell contamination than shorter copy-number variations. For the same size CNV, the detection ability of the deleted CNV was slightly higher than that of duplication. In the male sample, the observable shift of the X/Y chromosome ratio at >10% MCC can be detected by the microarray. Conclusions:When the MCC is higher than a particular ratio, it can affect the CMA detection of CNV. Based on the Agilent 180K CGH detection results for MCC mimics and the CNV detection specificity principle, the MCC threshold of the Agilent 180K CGH is set to 30% in view of conservative principles.

Prenatal diagnosis of mosaic trisomy 2: a case report / 中华围产医学杂志

We reported a case of mosaic trisomy 2.The patient was a 29-year-old gravida who underwent amniocentesis at 20 weeks of gestation because of high risk of trisomy-21 in the first trimester screening.The test result revealed a karyotype of 47,XN,+2[10]/46,XX[40].At 26 gestational weeks,the fetus was found severe fetal growth restriction and oligohydramnios which was considered to be at risk of mosaic trisomy 2.The pregnancy was terminated at 27+ gestational weeks.The fetus had obviously abnormal appearances,including dolichocephaly,low-set ears,and micromandible.Autopsy was not performed due to the parents' refusal.

An interlaboratory comparison study on the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels / 中华血液学杂志

Objective: To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison. Methods: Peking University People's Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients. Totally 23 sets with 14 different samples per set were prepared. TRIzol reagent was added in each tube and thoroughly mixed with cells for homogenization. Each laboratory simultaneously tested RUNX1-RUNX1T1 and WT1 transcript levels of one set of samples by real-time quantitative PCR method. All transcript levels were reported as the percentage of RUNX1-RUNX1T1 or WT1 transcript copies/ABL copies. Spearman correlation coefficient between the reported transcript levels of each participated laboratory and those of PKUPH was calculated. Results: ①RUNX1-RUNX1T1 comparison: 9 samples were (+) and 5 were (-) , the false negative and positive rates of the 20 participated laboratories were 0 (0/180) and 5% (5/100) , respectively. The reported transcript levels of all 9 positive samples were different among laboratories. The median reported transcript levels of 9 positive samples were from 0.060% to 176.7%, which covered 3.5-log. The ratios of each sample's highest to the lowest reported transcript levels were from 5.5 to 12.3 (one result which obviously deviated from other laboratories' results was not included) , 85% (17/20) of the laboratories had correlation coefficient ≥0.98. ②WT1 comparison: The median reported transcript levels of all 14 samples were from 0.17% to 67.6%, which covered 2.6-log. The ratios of each sample's highest to the lowest reported transcript levels were from 5.3-13.7, 62% (13/21) of the laboratories had correlation coefficient ≥0.98. ③ The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of WT1 transcript levels. Both RUNX1-RUNX1T1 and WT1 transcript levels from 2 and 7 laboratories were individually lower than and higher than those of PKUPH, whereas for the rest 11 laboratories, one transcript level was higher than and the other was lower than that of PKUPH. Conclusion: The reported RUNX1-RUNX1T1 and WT1 transcript levels were different among laboratories for the same sample. Most of the participated laboratories reported highly consistent result with that of PKUPH. The relationship between laboratories of the different transcript levels may not be the same.

Prenatal diagnosis and pregnancy outcomes of 22q11.2 duplication syndrome: analysis of 8 cases / 浙江大学学报·医学版

OBJECTIVE@#To investigate the relationship between 22q11.2 duplication and clinical phenotype.@*METHODS@#Eight fetuses with 22q11.2 duplication syndrome diagnosed by chromosome microarray analysis (CMA) through amniocentesis from February 2015 to March 2017 were enrolled in the study. The prenatal diagnostic indications, fetal ultrasound, chromosome karyotype, peripheral blood CMA results of parents, pregnancy outcomes and follow-up of postnatal growth and development were retrospectively analyzed.@*RESULTS@#Prenatal serological screening indicated 6 cases with high risk of trisomy 21, 1 case with nuchal fold (NF) thickening and 1 case of maternal chromosomal balanced translocation. Fetal ultrasonography showed 1 case of NF thickening, 1 case of fetal cerebral ventriculomegaly and 6 cases with normal ultrasound. CMA demonstrated that the size of duplication was between 651 kb and 3.26 Mb, and 22q11.2 duplication. Parents' CMA results revealed that 6 cases inherited from one of the parents with normal phenotype, and the parents of 2 cases refused the CMA test. Two couples chose induced labor; 6 cases of continued pregnancy had normal phenotypes at birth. All 6 cases were followed up with longest of 3.5 years. The growth and psychological development were normal in 5 cases, and one case was growth retardation.@*CONCLUSIONS@#There were no specific clinical phenotypes in 22q11.2 duplication syndrome, and most of them were inherited from one parent who has normal phenotype.

Analysis of the early immune effects of rabies vaccine combined with human rabies immunoglobulin / 中华实验和临床病毒学杂志

Objective@#To investigate the early immune effects of rabies vaccine combined with human rabies immunoglobulin (HRIG).@*Methods@#Rapid fluorescent focus inhibition test (RFFIT) was used to test the titers of rabies virus neutralizing antibodies (RVNA). The titers of RVNA of persons who had exposed to rabies at degree III on day 0, day 1, day 7, day 14 and day 45 were compared. The dynamic curves and seroconversion rates (SCR) of RVNA in persons of different genders, age and vaccine regimens (Essen and Zagreb) groups were analyzed.@*Results@#No significant differences of SCR among different genders, age and vaccine regimens groups were observed on the same day. SCR could be 100% on day14 in different groups. The dynamic curves of RVNA within the first 14 days showed the models of gradual rise, rapid rise and rapid decline.@*Conclusions@#The dynamic curves of RVNA within the first 14 days varied when rabies vaccines were used in combination with HRIG, and not all subjects were proved to be protected based on the RVNA detection within this period.

Pregnancy outcomes among women with high and low risk in first and second trimester prenatal screening analysis / 预防医学

Objective To analyze adverse pregnancy outcomes among women with high risk and low risk during prenatal screening. Methods Clinical data of 180006 pregnancies in 5 prenatal screening center in Hangzhou were collected. We compared the adverse pregnancy outcomes of high and low risk pregnancies. Results Among 180006 pregnancies (age<35 years old), there were 10296 high-risk cases and 169710 low-risk cases, with 168654 cases followed. There were 9406 high-risk cases of Down's syndrome (DS) (5.23％), 273 high-risk cases of Edwards' syndrome (ES) (0.15％) and 617 high-risk cases of open neural tube defect (ONTD) (0.34％) . The detection rate of pregnancy outcome of premature birth, spontaneous abortion, termination of pregnancy, stillbirth in the high risk was 5.46％, 0.80％, 1.80％, 0.37％, respectively, and that rate in low risk results was 3.50％, 0.21％, 0.38％, 0.18％, respectively. Adverse pregnancy outcomes in high-risk group were significantly higher than that in low-risk group (all P<0.001) . The actual incidence rate of DS, ES or ONTD in high-risk group (4.56‰, 1.65‰, 0.97‰, respectively) were significantly higher than that in low-risk group (0.12‰, 0.04‰, 0.09‰, respectively, all P<0.001) . The detection rates of prenatal screening were 70.15％, 68.00％ and 38.46％, and false positive rates were 5.23％ , 0.14％ and 0.34％ . Conclusion High risk of serum prenatal screening analysis is associated with adverse pregnancy outcomes. The number of pregnancies experienced invasive prenatal diagnosis can be reduced by routine serum prenatal screening. That is an Effective method to reduce the birth defects.

Observation on the efficacy of Conbercept for chronic central serous chorioretinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To observe the efficacy of intravitreal conbercept injection for chronic central serous chorioretinopathy (CSC).METHODS: Nine eyes of 9 patients diagnosed as chronic CSC between October 2015 to May 2016 were treated with an intravitreal injection of conbercept (0.5mg/0.05mL) (six patients were given the same does of intravitreal injection again at 1mo after the first injection).Follow-up observation was at 1, 2, and 6mo after injection.Observed indicators included best-corrected visual acuity (BCVA), intraocular pressure, optical coherence tomography (OCT), fundus fluorescein angiography (FFA), choroidal indocyanine green angiography (ICGA), macular fovea thickness (CMT), subfoveal choroidal thickness (SFCT).RESULTS:Seven of the 9 patients responded significantly to the drug, while 2 patients had no response.The CMT was 373.12±72.43μm at baseline, which decreased significantly to 332.05±67.13μm, 282.24±62.30μm and 225.56±71.08μm at 1, 2 and 6mo after the intravitreal injection.The mean thickness of SFCT was 422.11±64.82μm before treatment.The choroidal thickness of non-responsive patients before treatment was below average, respectively 353μm and 365μm.The SFCT of 1, 2, and 6mo after treatment was 391.45±75.24μm, 365.53±63.07μm, 355.40±66.65μm.Before treatment and 1mo after, there was no significant difference (P=0.074), but there was statistically significant (P0.05).CONCLUSION: Intravitreal conbercept injection in chronic CSC may have some effect in accelerating subertinal fluid resolution and decreasing the CMT.The SFCT within 6mo after treatment was significantly lower than pretreatment.The SFCT may be an indicator of whether patients respond.

Maternal serum alpha fetoprotein and free β-hCG of second trimester for screening of fetal gastroschisis and omphalocele / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To assess the detection of maternal serum alpha fetoprotein (MSAFP) and free beta-HCG levels of second trimester for screening of fetal gastroschisis and omphalocele.</p><p><b>METHODS</b>Clinical data of 622 639 pregnant women from 5 prenatal screening centers in Hangzhou during October 2007 and September 2016 were analyzed retrospectively. Thirty cases of gastroschisis and 30 cases of omphalocele diagnosed by ultrasonography and postmortem findings were enrolled in the study and 116 cases of pregnant women with normal fetal development during the same period were selected as control group. The cut-off value and area under ROC curve (AUC) of MSAFP and free β-hCG for diagnosis of fetal gastroschisis and omphalocel were analyzed.</p><p><b>RESULTS</b>MSAFP levels of women with fetal gastroschisis and omphalocele were 4.41 (0.88-11.69) MOM and 2.31 (0.72-23.20) MOM, which were significantly higher than that of control group[0.98 (0.41-2.26) MOM, all<0.01]. Free β-hCG level of women with fetal gastroschisis was 1.25 (0.35-19.94) MOM, which was significantly higher than that of control group[0.86 (0.17-6.11) MOM,<0.05). But there were no significant difference in free β-hCG between fetal omphalocele group[1.03(0.21-8.95)]and control group (>0.05). The AUCs of MSAFP for diagnosis of gastroschisis and omphalocele were 0.897 (95%:0.822-0.972) and 0.852(95%:0.762-0.942), respectively (all<0.01). Taking 1.655 MOM as the cut-off value of MSAFP for abdominal wall defects (gastroschisis and omphalocele), the sensitivity was 68.30%, specificity was 99.60% and Youden index was 0.649.</p><p><b>CONCLUSIONS</b>MSAFP of second trimester is a better biomarker than free β-hCG in screening abdominal wall defects.</p>

Study on the risk factors of chronic obstructive pulmonary disease in acute exacerbation complicated with pulmonary embolism / 国际检验医学杂志

Objective To analyze the risk factors of chronic obstructive pulmonary disease (COPD) in acute exacerbation (AE-COPD)complicated with pulmonary embolism ,and to provide reference for the pathogenesis and treatment .Methods A total of 73 patients with COPD suspected PE admitted to our hospital from May 2015 to April 2016 were enrolled in this study .All patients were examined including WBC ,Neu% ,CRP ,IL-8 ,ESR ,PCT ,ET-1 ,D-dimer ,fibrinogen ,NT-proBNP ,myocardial enzyme ,arterial blood gas ,lactic acid ,CT pulmonary angiography(CTPA)within 48 h of admission .The risk factors of AECOPD with PE or with-out PE were analyzed .Results There were 15 cases with PE ,58 cases without PE in all objects .Neu% ,PCT ,NT-proBNP ,D-di-mer ,LDH ,cTnI ,CRP ,IL-8 ,ET-1 in patients with PE were significant higher than those in patients without PE (P<0 .05) .In the PE group ,the correlation coefficient between CRP and IL-8 was 0 .457(P=0 .087) ,the correlation coefficient between CRP and ET-1 was 0 .598(P=0 .019) ,the correlation coefficient between IL-8 and ET-1 was 0 .695(P=0 .004) .Conclusion Acute exacer-bation of COPD combined with PE is associated with the severity of inflammation in the body ,the more serious the inflammatory re-action ,the corresponding increase in myocardial injury ,the higher the risk of PE .

Study on the risk factors of chronic obstructive pulmonary disease in acute exacerbation complicated with pulmonary embolism / 国际检验医学杂志

Objective To analyze the risk factors of chronic obstructive pulmonary disease (COPD) in acute exacerbation (AE-COPD)complicated with pulmonary embolism ,and to provide reference for the pathogenesis and treatment .Methods A total of 73 patients with COPD suspected PE admitted to our hospital from May 2015 to April 2016 were enrolled in this study .All patients were examined including WBC ,Neu% ,CRP ,IL-8 ,ESR ,PCT ,ET-1 ,D-dimer ,fibrinogen ,NT-proBNP ,myocardial enzyme ,arterial blood gas ,lactic acid ,CT pulmonary angiography(CTPA)within 48 h of admission .The risk factors of AECOPD with PE or with-out PE were analyzed .Results There were 15 cases with PE ,58 cases without PE in all objects .Neu% ,PCT ,NT-proBNP ,D-di-mer ,LDH ,cTnI ,CRP ,IL-8 ,ET-1 in patients with PE were significant higher than those in patients without PE (P<0 .05) .In the PE group ,the correlation coefficient between CRP and IL-8 was 0 .457(P=0 .087) ,the correlation coefficient between CRP and ET-1 was 0 .598(P=0 .019) ,the correlation coefficient between IL-8 and ET-1 was 0 .695(P=0 .004) .Conclusion Acute exacer-bation of COPD combined with PE is associated with the severity of inflammation in the body ,the more serious the inflammatory re-action ,the corresponding increase in myocardial injury ,the higher the risk of PE .

The Efficacy and Prognostic Factors for Long Pulse Neodymium: Yttrium-Aluminum-Garnet Laser Treatment on Onychomycosis: A Pilot Study

No abstract available.

Characterization of Serial Passage of 1b/2a Chimera Hepatitis C Virus Cell Culture System Carrying Envelope E1E2 Coding Gene from Hebei Strain of China / 病毒学报

To character a novel chimera(1b/2a) hepatitis C virus cell culture (HCVcc) system carrying envelope E1E2 coding gene from Hebei strain of China, chimera HCVcc (cHCVcc) was developed from Huh7.5-CD81 cells after transfection with in vitro transcribed full-length 1b/2a chimera RNA, which carrying envelope E1E2 coding gene from Hebei strain of China. Then the replication, expression and infectious titer of serial passage HCVcc were assessed by Real Time RT-PCR, indirect immunofluorescence assay (IFA) and Western blotting (WB). In addition, chimeric envelope gene from HCVcc was sequenced after serial passage. We found that the number of HCV positive focus increased gradually in cell post-transfection with chimera HCVcc (1b/2a) RNA and reach a peak platform (80% to 90%) at 41 days post-transfection; the expression of HCV protein was also confirmed by WAB during serial passage. At meantime, HCV RNA copy number in the supernatant peaked at 10(4)-10(7) copies/mL and the highest infectious titer of this 1b/2a cHCVcc reinfection were tested as 10(4) ffu/mL. Sequence analysis indicated 6 of adaptive amino acid substitutes occur among chimeric envelope E1E2 during serial passages. We con:luded that a novel 1b/2a chimera HCVcc carrying envelope E1E2 coding gene from Hebei strain of China was developed and its infectious titer increased after serial passage of HCVcc. This novel cHCVcc will be an effective tool for further evaluation of anti-virus drugs and immune effects against the major genotype from Chinese.

Roles of complement receptor 3 on murine macrophages in recognition of Penicillium marneffei / 中华皮肤科杂志

Objective To evaluate the role of complement receptor 3 (CR3) on murine macrophages in the recognition of Penicillium marneffei.Methods RAW264.7 murine macrophage cells were cultured in vitro,and divided into four groups to be cocultured with inactivated and live Penicillium mameffei yeast cells as well as inactivated and live Penicillium marneffei conidia respectively at 37 ℃ in 5％ CO2 for one hour.The RAW264.7 cells incubated with phosphate-buffered saline (PBS) served as the blank control group.Then,reverse transcription-PCR was conducted to detect CR3 mRNA expression,Western blot to measure CR3 protein expression,flow cytometry to determine phagocytosis rate,enzyme-linked immunosorbent assay (ELISA) to quantify cytokine levels in culture supernatant.Some RAW264.7 macrophages were transfected with a specific siRNA targeting CR3 gene and cocultured with inactivated Penicillium marneffei conidia,subsequently,phagocytosis rate and supematant cytokine levels were determined.Data were processed by the SPSS 16.0 software,and one-way analysis of variance (ANOVA) was conducted for inter-group comparisons of these parameters.Results No significant differences were observed in the mRNA or protein expressions of CR3 among the four groups of RAW264.7 cells cocuhured with different forms of Penicillium marneffei (both P ＞ 0.05).The phagocytosis rate was 95.14％,89.56％,91.03％ and 90.78％ in RAW264.7 cells cocultured with inactivated conidia and yeast cells,as well as live conidia and yeast cells of Penicillium marneffei,respectively (P ＞ 0.05).The levels of interleukin (IL)-2,interferon (IFN)-γ,IL-4 and IL-10 in culture supernatant were increased at different degrees after one-hour coculture in the four coculture groups compared with the blank control group,but no statistical difference was noted among the four coculture groups in the supernatant levels of these cytokines (all P ＞ 0.05).After coculture with inactivated Penicillium marneffei conidia,the siRNA-transfected RAW264.7 cells showed a statistical decrease in phagocytosis rate (10.89％ vs.92.78％,P ＜ 0.05) and supernatant levels of IL-2,IFN-γ IL-4 and IL-10 compared with untransfected RAW264.7 cells.Conclusions In early stage of innate immunity,CR3 on macrophages may be one of the pattern recognition receptors participating in the recognition and mediation of phagocytosis of Penicillium marneffei.It's possible that both Thl-and Th2-type cytokines,such as IL-2,IFN-γ,IL-4 and IL-10,are involved in the immune response of macrophages against Penicillium marneffei.

Renovation stem revision femoral head arthroplasty for unstable intertrochanteric fracture in the elderly / 中国骨伤

<p><b>OBJECTIVE</b>To observe clinical efficacy of renovation stem revision femoral head arthroplasty for the treatment of unstable intertrochanteric fracture in the elderly.</p><p><b>METHODS</b>Totally 32 elderly patients with unstable intertrochanteric fracture were treated with renovation stem revision femoral head arthroplasty from September 2007 to January 2011. There were 11 males and 21 females with an average age of 83.8 (ranged, 80 to 98) years old,the time from injury to hospital ranged from 4 h to 14 days. According to Evans-Jensen classification, 6 cases were type II a, 20 cases were type II b, and 6 cases were type III. Postoperative mortality, complication rates and Harris hip function score were compared and analyzed to evaluate curative effect.</p><p><b>RESULTS</b>All patients were followed up and no dislocation occurred. Six patients were died during 15 months and 4.5 years; 24 cases recoved to independent wakling at 6 months after operation, and 8 cases walked with stick and walker. The average Harris hip joint function score were (91.56 +/- 2.96), 28 cases got excellent results and 4 cases good. Nine cases occurred complications and healed after treatment.</p><p><b>CONCLUSION</b>Renovation stem revision femoral head arthroplasty is a active and reliable method in treating unstable intertrochanteric fracture in the elderly.</p>