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Accurate detection of low frequency mutations from plasma cell-free DNA in blood using targeted next generation sequencing technology has shown promising benefits in clinical settings. Duplex sequencing technology is the most commonly used approach in liquid biopsies. Unique molecular identifiers are attached to each double-stranded DNA template, followed by production of low-error consensus sequences to detect low frequency variants. However, high sequencing costs have hindered application of this approach in clinical practice. Here, we have developed an improved duplex sequencing approach called SinoDuplex, which utilizes a pool of adapters containing pre-defined barcode sequences to generate far fewer barcode combinations than with random sequences, and implemented a novel computational analysis algorithm to generate duplex consensus sequences more precisely. SinoDuplex increased the output of duplex sequencing technology, making it more cost-effective. We evaluated our approach using reference standard samples and cell-free DNA samples from lung cancer patients. Our results showed that SinoDuplex has high sensitivity and specificity in detecting very low allele frequency mutations. The source code for SinoDuplex is freely available at https://github.com/SinOncology/sinoduplex.
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Objective To investigate the changes of Caprini score, plasma D-D and FIB levels in patients with post-fracture DVT and its diagnostic significance.Methods Sixty patients with post-fracture DVT (DVT group) and 120 patients without DVT after fracture were selected (control group).The Caprini score, plasma D-D and FIB levels were compared between the two groups.The relationship between Caprini score and plasma D-D and FIB in the DVT group was analyzed.Results The Caprini score, D-D and FIB levels in the DVT group[7.26±1.88, (3.31±1.52) mg/L, (4.48±0.72) g/L] were significantly higher than those in the control group[3.39±0.80, (0.46±0.11) mg/L, (3.60±0.68) g/L], and the difference was statistically significant (P＜0.05).There was a significant positive correlation between Caprini score and D-D and FIB levels in patients with DVT (P＜0.05).There was a positive correlation between Caprini score and level of D-D and FIB (r=0.748、r=0.662, P＜0.05).The sensitivity of Caprini score combined with D-D and FIB levels in the diagnosis of DVT in fracture patients was 97.83％, specificity was 92.00％, missed diagnosis rate was 2.17％, misdiagnosis rate was 8.00, and area under ROC curve was 0.947 (P＜0.05).Conclusion Caprini score, D-D and FIB levels in patients with lower limb fracture have certain clinical value for the diagnosis of DVT.
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Objective To understand the development history and current situation of healthcare-associated infection(HAI)management in children’s hospitals in China during the past 30 years.Methods A retrospective survey was conducted in 20 children’s hospitals,the investigated contents included:basic situation of hospitals, HAI management organizations and professionals,the support of hospital leaders for the management of HAI, training on HAI knowledge,monitoring on HAI,and so on.Results Up to 2015,20 hospitals all set up independent HAI management departments and HAI management committees,46.15%(6/13)of hospitals with more than 1 000 beds were understaffed in HAI management.There were a total of 88 professionals in HAI management in 20 children’s hospitals in 2015,53.41% (n=47)of professionals had bachelor degree,34.09% (n=30)were with intermediate professional title,46.59%(n=41)were nursing staff.Up to 2015,incidences and missing report rates of HAI,antimicrobial usage rates,incidences of HAI in neonates all decreased compared with 2006.In 2015,20 (100.00%)hospitals all carried out monitoring on antimicrobial application,multidrug-resistant organisms,disin-fection and sterilization,and provided personnel protective equipment.In 2015,9(45.00%)hospitals used non-tou-ch faucets in all wards,19(95.00%)hospitals supplied hand-drying paper towel and alcohol-based hand rub in all wards.None of professionals in these hospitals participated in national-level training on HAI for ≥3 times in 1996, professionals in 11 hospitals (55.00%)participated in national-level training for ≥3 times in 2015.Conclusion In the past 30 years,the management of HAI in children’s hospitals in China has basically met the requirement,moni-toring on HAI gradually improved,but allocation of HAI professionals and personnel quality ability still need to be strengthened.
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Objective To analyze effect of the syphilis prevention and control work among female sex workers(FSWs), and provide reference data for target intervention in Guangdong province.Methods Syphilis prevalence among FSWs from 2009 to 2013 were analyzed statistically based on the data from Guangdong Provincial high-risk groups in sexually transmit-ted infection(STI)control program.Results A total of 34 338 FSWs were detected,including 23 816(69.36%)FSWs at the high-tier venues,8 811(25.66%)middle-tier,and 1 711(4.98%)low tier.The syphilis prevalence of FSWs at high-, middle-,and low-tier venues in 2009 was 2.60%,7.36%,and 10.54% respectively,and in 2013 was 1.45%,2.75%,and 21.88% respectively.Chi-square test showed a downward trend in the prevalence of syphilis among the high and middle tier FSWs in 2009-2013(χ2 =12.807,52.333 respectively,both P <0.001),and remained high among FSWs at low-tier venues(χ2 =0.027,P =0.87).The difference in syphilis prevalence in 2013 among FSWs from different types of venues was statistically significant (χ2 =190.64,P <0.001 ).No significant difference in syphilis prevalence was found among FSWs from Pearl River Delta,eastern,western and northern Guangdong(χ2 =5.24,P =0.07).Conclusion Significant difference is found in prevalence of syphilis among FSWs from different types of settings.FSWs from low-tier settings have a relatively lower syphilis testing rate but with higher infection rate.
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OBJECTIVE To explore the role of runt-related transcription factor 3(RUNX3) in the tumorgenesis and progression of cervical carcinoma. METHODS The immunohistochemical staining technique was used to detect the expression of RUNX3 protein in 25 cases of normal cervix, 34 intraepithelia neoplasia (CIN), and 48 cervical carcinomas. SYBR Green I chimeric fluorescence Real-time PCR was applied to detect the expression of RUNX3 mRNA in 10 cases of normal cervix, 24 CIN, and 30 cervical carcinomas. RESULTS The expressions of RUNX3 protein and mRNA in normal cervix, CINI,CINII-III, and cervical carcinoma tissues tended to be down-regulated. There was significant difference among these groups (P 0.05). CONCLUSION RUNX3 may function as a tumor suppressor gene in the occurrence and progression of cervical carcinoma.
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Adult , Female , Humans , Middle Aged , Carcinoma, Squamous Cell , Genetics , Metabolism , Uterine Cervical Dysplasia , Genetics , Metabolism , Core Binding Factor Alpha 3 Subunit , Genetics , Metabolism , Disease Progression , RNA, Messenger , Genetics , Metabolism , Uterine Cervical Neoplasms , Genetics , MetabolismABSTRACT
Objective: To probe into the relationship between integrin β1 expression in human cervical carcinoma HCE1 multicellular spheroids (HCE1/MCS) and their invasion into human umbilical vein endothelium cells (HUVEC), so as to assess the inhibitory effect of anti-integrin β1 monoclonal antibody P5D2 on the invasion of HCE1/MCS into HUVEC. Methods: A model of HUVEC monolayer invaded by HCE1/MCS was established (in brief, the HCE1 invading model). Morphology changes of the HCE1 invading model were observed under inverted microscope and analyzed by Motic Med System after P5D2 treatment. The expressions of β1 integrin on HCE1 monolayer cells, HCE1/MCS, HVUEC monolayer cells, and P5D2-treated HCE1 invading models were measured by immunocytochemistry SABC assay. Results: A HCE1 invading model was successfully established. The HCE1/MCS proliferated rapidly after culture, and on the 7th day the invading area of HCE1/MCS was 40.42 folds larger than the original HCE1/MCS. Invading areas of P5D2-treated HCE1/MCS were significantly smaller than that of the control group after 1, 4, 7 day (P<0.05, or P<0.01), with the invading area after 7 days reduced by (84.68±0.08) % compared with the control group. Integrin β1 expression in HCE1/MCS was significantly higher than that in HCE1 monolayer cells (P<0.01), and the expression was negative in HUVEC. Integrin β1 expression in P5D2-treated HCE1/MCS was significantly lower than that the in untreated HCE1/MCS (P<0.01). Conclusion: Upregulated expression of integrin β1 in HCE1/MCS and HCE1 invading model may be associated with their enhanced adhesion and invasion abilities. Anti-integrin β1 monoclonal antibody P5D2 can partially block the invasion of HCE1/MCS into HUVEC.
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Objective To investigate expression of extracelluar signal regulated protein kinase(ERK)and phosphorylation ERK(p-ERK)in the temporal lobe from patients with DR-TLE so as to explore the possible roles of ERK in the pathogenesis of DR-TLE.Methods Expression of ERK was detected with Western blot and immunohistochemistry in 32 patients with DR-TLE(24 temporal lobe,8 hippocampi),as compared with 12 controls(9 temporal lobe,3 hippocampi).Results ERK and p-ERK expression in DR-TLE was significantly higher(0.2266?0.0613,0.2097?0.0183 and 0.1924?0.0054,respectively)than those of controls(0.1840?0.0023,0.1974?0.0056 and 0.1825?0.0063,respectively,all P
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Objective To study the expression of heat shock 27 000 associated protein 1 ( HSPBAP1, GenBank: AK096705) in the brain tissues of patients with drug-refractory epilepsy and discuss its function in the pathogenesis. Methods Fluorescent quantitative polymerase chain reaction ( FQ-PCR) and immunohistochemistry were used to test the expression of HSPBAP1 in the surgically removed brain tissues of patients with drug-refractory epilepsy from the brain bank of our department ( n = 36) , and the results were compared with that of normal controls (n = 8 ). Results The relative expression of HSPBAP1 mRNA in the brains of patients with drug-refractory epilepsy was more than 34. 11 times that of controls, and HSPBAP1 protein expression was significantly increased in temporal lobe cortex (0. 0507?0. 0003, P
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AIM：Damaging of Endothelial cells and adhesion of leukocytes and platelets were studied in rat mesentery microvessels after ischemia-reperfusion.METHODS：The model was made by losing blood and reperfusion from carotid artery in rat. Changes in mesentery microvessels were observed by high magnify microscope.RESULTS:Leukocyte and platelet adhesions were found in venules and co-capillaries 1-3 hours after ischemia-reperfusion. Endothelial cells were edema and vascular walls were thickening. Vacuoles formed in intracytoplasm of some vascular endothelium and some bigger endothelial vacuoles prominenced toward the luminal surface. Vacuoles had the shape of circle and the diameter was 10-30 μm. More vacuoles were found in arterioles, even there were several vacuoles in a arteriole. The biggest vacuole almost occupied 2/3 of vascular lumen.CONCLUSION：Edema and vacuole formation in vascular endothelium showed endothelial cells were damaged seriously after ischemia-reperfusion.
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0.05).Follow-up was available for(14.2?3.6) months(range,7-19 months) in the laparoscopic group and for(13.1?4.2) months(range,7-19 months) in the laparotomy group.No recurrence was detected in both the groups during the follow-up.ConclusionLaparoscopic surgery do not affect the expression levels of the factors,including E-cadherin,MMP-2,VEGF-C,and CD44v6,that influence the metastasis and implantation of tumor cells in patients with endometrial cancer.
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Objective This study was to investigate the Chlamydia trachomatis (CT) infection rate in cervical secretion of early pregnant and secondary sterility women. Method CT-DNA in the cervical secretion of the early pregnant, the secondary sterility, and the control women was detected with polymerase chain reaction (PCR). Results The infection rates of CT in the early pregnant and secondary sterility women （14.28% and 13.33%）were higher than in the control women (3.33%, P<0.05),and there was no significant difference between the former two groups (P>0.05). Conclusion There are higher infection rates in the early pregnant and secondary sterility women.Detecting CT and treating in the perinatal period are needed in the early pregnant women who want to be pregnant again.
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Objective To investigate the expression of cyclin dependent kinases 5(CDK5)in the temporal lobes of the epilepsy patients and to explore the possible roles of CDK5 in the pathogenesis of refractory epilepsy.Methods The brain tissues of intractable epilepsy(IE)were studied by fluorescence quantative polymerase chain reaction(FQ-PCR)for CDK5 mRNA,while immunohistochemistry and Western blot were used to study the protein expression.Nonepileptogenic control brain tissues were used for comparison.Results FQ-PCR analysis showed that the expression of CDK5 mRNA in epilepsy patients was significant higher than those in the control group.And immunohistochemistry showed that the protein mainly existed in the neuron and glial.At the 35000 relative molecular mass,Western blot could been seen that there is a limpid strap.The optical density of CDK5 in IE(temporal lobe 1.4293?0.1839,hippocampus 2.0733?0.4738)was significantly higher than that in the control(temporal lobe 0.9680?0.4147, hippocampus 1.4030?0.6160,P
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Objective To investigate on out the deformity in the elderly of longevous area and the characteristics of the regions in Yunnan Province. Methods According to the forth census information, using multistage sample typical survey to the 18 counties (cities) with centenarians over 1/100 000. The deformity types of people aged 60 and over and the deformity rate of various age groups and its distribution were investigated. Results Among the 6 477 people aged 60 and over investigated, a total of 1 928 people were in deformity, the deformity rate was 29 8%. The elder the people, the higher the deformity rate was. The highest deformity rate was in Kunming and Chuxiong prefectures. The audition deformity was highest among all types of deformity in elder people. The occurrence of fracture was highest in Miao Nation in Honghe prefecture, reaching 18.1%. About 14 % of the centenarian people sufferred from dementia. Conclusions Effective control in blood pressure and osteoporosis are important method to protect the elder people and to promote the level of quality of life of the elderly.
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AIM: To investigate the effects of human urotensin II (hUII) on ischemia/reperfusion (I/R) injury in isolated rat hearts. METHODS: In the ischemia/reperfusion (I/R) model of isolated perfused rat hearts,the effects of hUII pretreatment on cardiac function was monitored with cardiac function software of MFL Lab200. ATP,total calcium,and malondialdehyde (MDA) content in myocardium were detected. The coronary perfusion flow (CPF) and lactate dehydrogenase (LDH) activity in coronary effluent were measured during reperfusion. RESULTS: In the hUII pretreated group,the release of LDH from myocardium was lower [(78.3?18.1)U/L] than I/R group [(109.3?23.9) U/L,P
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AIM: To investigate the regulatory effects of salusins on tensility of microvessle in rats. METHODS: Wistar rats were randomly distributed into six experimental groups as follows: (1) normal saline injection-treated control; (2) salusin ?; (3) salusin ?; (4) noradrenaline; (5) noradrenaline+salusin ?; and (6) noradrenaline+Salusin ?. For recording the microcirculation images in the mesentery, the intestinal loop was mounted on the stage of an intravital microscope equipped with a TV camera. Temporal changes in internal diameter of arterioles and venules were measured by computer using the Image-Pro software. RESULTS: Salusin ? (10 -5mol/L) dilated arterioles directly and the dilation up to the peak at 3 min [(25.56?4.30) vs (24.49?4.27)?m, P0.05). Salusin ? has no effects on internal diameter directly and NA-induced contraction of microvessles. CONCLUSION: Salusin ? dilated mesenteric arterioles and inhibited NA-induced contraction of microvessles in rats.
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AIM: To investigate the effects of human urotensin Ⅱ (hUII) on in vivo mesenteric microcirculation in rats. METHODS: For recording of microcirculation images in the mesentery, the intestinal loop was mounted on the stage of an intravital microscope equipped with a TV camera. Video images of microcirculation were stored by a video cassette recorder. Temporal changes in internal diameter and microcirculatory velocity of microvesseles were measured by computer using the ImagePro software. The blood flow in intestinal wall was measured with PIMII laser Doppler perfusion Imager (Lisca Sweden). RESULTS: The internal diameters of arterioles and venules in control group were (21.4?2.3) ?m and (38.1?3.6) ?m,respectively. In UII group, the arterioles and venules contracted immediately after treated with UII and up to the peak at 1 min [(14.1?1.4) ?m and (22.2?5.2) ?m vs control,P0.05). The blood flow in intestinal wall increased 1 min after treated with UII and up to high peak at 5 min(6.4?1.1 perfusion unit vs control 4.2?0.9,P
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AIM: To investigate the effects of hypoxia-inducible factor-1 (HIF-1) on cardioprotection of hypoxic preconditioning (HPC) and its mechanisms. METHODS: In the model of hypoxia/reoxygenation (H/R) of cardiomyocytes from neonatal Spargue-Dawley rats, delayed cardioprotective effects of HPC on lethal H/R were observed. Whole cell extracts were prepared for determining activities of extracellular signal-regulated protein kinases (ERKs) and HIF-1? phosphorylation. RESULTS: HPC significantly promoted survival and membrane integrity of cardiomyocytes subjected to sustained H/R. SDS-PAGE mobility shift experiments showed increased phophorylation level of HIF-1? in hypoxic preconditioned cardiomyocytes. ERK1/2 were activated at 10 min after HPC and returned to the control level at 60 min. CONCLUSION: HPC protects neonatal cardiomyocytes from H/B injury. HIF-1? phosphorylation induced by ERKs might contribute to the cardioprotection of HPC.
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AIM:To study whether ischemic preconditioning(IPC) has a protective effect against ischemia/reperfusion(I/R) injury in brain, and the possible relationship between IPC and the regulating function of microcirculation. METHODS: The I/R models were established both in I/R and IPC groups of Sprague-Dawley rats. Additional procedure was performed of short term cerebral ischemic preconditioning in IPC group 24 hours before I/R. Skull windows were performed through which microcirculation features were measured before ischemia, during ischemia, and reperfusion. Finally, brains were cut into slices and stained with red tetrazoline(TTC). RESULTS: Most TTC stained brains in I/R group presented irregular palely red areas which were few in IPC group. Compared with I/R group, IPC group presented relatively increase in accumulated length of capillaries, mean cerebral microcirculatory perfusion, and microcirculatory velocity in ischemic and reperfusion phase. There was no-reflow phenomenon in I/R group in reperfusion phase, which was substituted by the course of increasing reperfusion in IPC group. CONCLUSIONS:IPC could relieve the reduction of tissue perfusion during ischemia and the no-reflow phenomenon during reperfusion by improving the regulating function of microcirculation, which relatively promote the opening of capillaries and accelerating of microvascular flow, therefore protect brain from I/R injury.