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Rapid, sensitive and specific detection tools are critical for the prevention and control of infectious diseases. The in vitro nucleic acid amplification assays, including polymerase chain reaction and isothermal amplification technology, have been widely used for the detection of pathogens. Recently, nucleic acid detection-based on clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) have been developed, which are rapid, highly sensitive, highly specific, and portable. This review describes the classification and principle of CRISPR/Cas systems and their applications in pathogen detection, and discusses the prospects of CRISPR/Cas systems.
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【Objective】 To explore the safety and efficacy of a novel endoscopic two-wire guided dilation in the creation of channels in percutaneous nephrolithotomy (PCNL). 【Methods】 Clinical records of 180 patients undergoing PCNL during Oct.2020 and Oct.2022 were retrospectively analyzed. The patients were divided into three groups, 60 in AMD group (fascial amplatz dilation), 60 in OSD group (one shot dilation) and 60 in END group (endoscopic dilation). Time to establish channels, operating time, failure of access, stone clearance rate, drop in hemoglobin, embolization rate, fever rate, blood transfusion rate and postoperative hospitalization were compared among the three groups. 【Results】 There were no significant differences in the general data among the three groups (P>0.05). Compared with AMD and OSD groups, END group needed significantly reduced time to establish the first channel [(5.6±0.8) min vs. (4.9±1.4) min vs. (4.2±0.5) min, (P<0.05)] . Compared with OSD group, END and AMD groups had significantly more hemoglobin drop [(14.0±17.6) g/L vs. (19.4±12.6) g/L vs. (10.2±6.8) g/L, (P<0.05)] . There were no significant differences in terms of failure of establishing channels, operating time, stone clearance rate, embolization rate, fever rate, blood transfusion rate and postoperative hospitality. Four patients needed selective renal artery embolization (1 case in AMD group and 3 in OSD group). No serious complications such as organ injuries, septic shock or death occurred. 【Conclusion】 Endoscopic two-wire guided dilation is simple, with few complications and good application value.
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Objective:To identify the expression level of known microRNA (miRNA) by high-throughput sequencing and analysis of the miRNA of Schistosoma japonicum schistosomula, and predict the miRNA target genes and their biological functions. Methods:Schistosoma japonicum schistosomula were prepared in vitro, and total RNA of schistosomula were extracted and analyzed to construct a library for performing high-throughput sequencing. The difference of miRNA expression was analyzed by using DEGseq R language package and perl script. Then the target genes and their biological functions of differential miRNA were predicted by miRanda software, Blast software, and KEGG database respectively. Results:There were 38 483 matching sequences in the miRNA expressed of Schistosoma japonicum schistomula in the constructed library compared with the latest miRBase database, and 60 miRNA were identified, sja-miR-125b was the miRNA with the highest expression, followed by sja-miR-61, sja-miR-71a, sja-miR-36-3p and sja-miR-10-5p, which accounted for 91% (3 263/3 585) of the total miRNA expression. MiRanda software predicted a total of 7 176 target genes, gene functions concentrated on nucleotide transferase activity, cellular nitrogen complex metabolism, molecular function, biological processes, biosynthesis, plasma membrane and protein maturation. The functional enrichment analysis showed that the highly expressed miRNA were mainly involved in pathogenic process, biological progress and multiple metabolic regulation pathways. Conclusions:The miRNA expressed significantly of Schistosoma japonicum schistomula mainly involved in the regulation of metabolic pathways during the differentiation, growth and pathogenesis of Schistosoma japonicum. To lay a foundation for the study of the regulatory mechanism of Schistosoma japonicum development and the development of new drugs.
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Objective T o explore the application value of postm ortem com puted tom ography (C T ) an-giography on diagnosis of coronary atherosclerotic stenosis degree. Methods B ased on the previous ex-perim ental results, the postm ortem C T angiography device of hum an isolated heart w as im proved. D iffer-ent coronary atherosclerotic stenosis degree of sudden death cases w as selected. B efore the cardiac anatom y, hearts w ere rem oved out com pletely and C T angiography w as perform ed im m ediately. T he C T angiography results w ere com pared w ith histopathological findings. M eanw hile, the advantages and disad-vantages of the angiography device before and after im provem ent w ere com pared. Results T he im proved angiography device of isolated heart could get better im aging results. T he postm ortem C T angiography results had high consistency w ith the histopathological findings on diagnosis of coronary atherosclerotic stenosis degree. A nd the coronary artery lesions could be revealed m ore objectively and vividly by 3D reconstruction technology. H ow ever, C T angiography could only be used to exam ine the pathological changes of blood vessels, w hich m ight have som e lim itations on the diagnosis of cause of death. Con-clusion Postm ortem C T angiography can be used as an additional m ethod for the conventional autopsy in the cases of coronary atherosclerosis.
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Objective To produce a comprehensive transcript dataset of Oncomelania hupensis before and after Schistosoma japonicum infection,so as to provide experimental data for perfecting genetic structural information and excavating related mo-lecular markers of O. hupensis infected by S. japonicum. Methods O. hupensis snails were divided into the following 3 groups:one week after S. japonicum miracidium infection,4 weeks after S. japonicum miracidium infection,and normal condition. Mil-lion high-quality reads were obtained from the normalized cDNA of the pooled samples,which were assembled into transcripts. Results A total of 63686 unigenes were identified and were classified into 4 main categories,including general functional pre-diction(15.36%),signal transduction mechanism(11.75%),posttranslational modification(8.89%),and functional unknown (12.20%). Conclusions The transcriptome information of O. hupensis snail after the invasion of S. japonicum shows that sever-al genes are significantly up-regulated or down regulated expression,and that the availability of transcriptome information might provide a strong foundation for further understanding the schistosome-snail interaction at the molecular level.
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Objective To discuss the relationships between regular pattern changes of plasma fibrinogen (Fib),D-dimer and fibrinogen degradation products (FDP) levels and the recent dissolution of thrombus in patients with pulmonary embolism (PE) in 14 days after treatment.Methods A prospective study was conducted.PE patients admitted to Departments of Respiratory Disease in 4 hospitals from January 2015 to March 2016 were enrolled and all of them were treated with thrombolysis and/or anticoagulation after admission.The computed tomographic pulmonary angiography (CTPA) was examined pre-treatment and 14 days post-treatment in PE patients.The pulmonary artery obstruction index (PAOI) was assessed according to the Mastora scoring method to estimate the thrombus load.The plasma Fib,D-dimer and FDP levels were measured before and on 1,2,3,5,7 and 14 days after treatment,and the relationships between the change regularities of these parameters and PAOI were also analyzed.Results A total of 42 PE patients were enrolled.The curve change of coagulation-fibrinolytic system parameters in 14 days after treatment showed that the Fib level was raised to its peak on the 3rd day after treatment and then decreased (g/L:4.24 ± 1.45 vs.3.83 ± 1.56),representing that its curve change was in accordance with the quadratic model (P =0.095).After treatment,the D-dimer and FDP levels were kept declining,they were reached the valley on 14th day [D-dimer (mg/L):1.58 ± 1.38vs.8.84 ± 6.35,FDP (mg/L):4.23 ± 3.63 vs.23.41 ± 16.54],and their curve changes were in accordance with the cubic model (F was 32.190 and 34.326,respectively both P =0.000).The PAOI variation before and 14 days after treatment [(18.77 ± 14.22)％] was not correlated with Fib variation [(1.20 ± 0.93) g/L,r =-0.194,P =0.219],but was positively correlated with D-dimer variation [(7.29 ± 7.10) mg/L] and FDP variation [(19.29 ± 18.67) mg/L,r was 0.556 and 0.460,respectively;P was 0.020 and 0.002,respectively].Conclusions The D-dimer and FDP levels are kept falling in PE patients after treatment,suggesting that the pulmonary artery embolus is being dissolved.
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Objective To explore the application value of postm ortem m ulti-slice spiral com puted tom o-graphy (MSCT) by observing and analyzing the injury features in the traffic accident victim s. Methods Ten traffic accident victim s w ere scanned w ith w hole body MSCT. The system ic autopsy w as subse-quently perform ed to com pare w ith the results of MSCT. The advantages and disadvantages of autopsy and MSCTfor obtaining the inform ation of traffic accident injuries w ere then analyzed. Results MSCTcould reveal 3D shape of fractures clearly and detect air accum ulation in different positions of the body, w hich show ed the obvious advantages com pared w ith autopsy. H ow ever, the resolution of MSCTw as lim ited com pared to the detection of organ and soft tissue injuries. Conclusion A com bination of MSCTand autopsy is the best w ay for determ ining the m anner and the cause of death in traffic fatality victim s.
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Objective To investigate the effect of nursing with solution focused approach in advanced schistosomiasis pa?tients. Methods Fifty three patients with advanced schistosomiasis in the Liujiahu Hospital for Schistosomiasis Control of Yiyang City were randomized selected and separated into an intervention group and a control group. The patients of the interven?tion group received the conventional nursing plus solution focused approach,while the patients of the control group received on?ly the conventional nursing. The effectiveness of nursing and treatment was evaluated in the two groups comparatively. Results The knowledge of self?care skills,medical compliance behaviors and services satisfaction degree in the intervention group were higher than those in the control group(c2=3.78,2.87,4.09 respectively,all P<0.05). The occurrence rate of upper gastroin?testinal bleeding in the intervention group was significantly lower than that in the control group(c2=4.894,P<0.05). The aver?age hospitalization duration of the intervention group was shorter than that of the control group(t=4.17,P<0.05). Conclu?sions The solution focused approach is a feasible and effective method in course of nursing of advanced schistosomiasis pa?tients. It enhances the confidence of the patients and the trustiness of the patients to the health care providers. The complications and the hospitalization duration of the patients are reduced.
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Postm ortem chem istry is becom ing m ore and m ore essential in routine forensic pathology and has m ade considerable progress over the past years. B iochem ical analyses of vitreous hum or, blood, urine and cerebrospinal fluid m ay provide im portant inform ation in determ ining the cause of death or in elucidating forensic issues. Postm ortem chem istry m ay be essential for the determ ination of cause of death w hen m orphological m ethods (diabetes m ellitus, alcoholic ketoacidosis and electrolytic disorders) cannot detect the pathophysiological changes involved in the death process. It can also provide m any in-form ation in other forensic situations, including m yocardial ischem ia, sepsis, inflam m ation, infection, ana-phylaxis and horm onal disturbances. T he m ost recent relevant research advances on glucose m etabolism , liver function, cardiac function, renal function, sepsis, inflam m ation, infection, anaphylaxis and horm onal aspect are hereby review ed.
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In drowning cases, it is difficult to judge whether the deceased died from drowning or throw-ing into the water after killed or identificate the cause of death of the decom posed corps in forensic practices. The diatom test is still considered as the im portant assistant evidence in drowning cases. This paper reviews research progress on technologies in recent years of forensic diatom test, and the applica-tion value of the newdeveloping approach in the field of forensic m edicine.
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Objective To explore the application value of multi-slice spiral computed tomography (MSCT) in traffic accidents through observing and analyzing the injury features of the accidents. Methods Two fatal cases caused by traffic accidents were fully examined using MSCT, 3D imaging reconstruction and angiography through cardiac puncture. The features of traffic injury mechanism were analyzed through combination of MSCT and postmortem external examination. Results In case 1, right cardiac rupture was found by MSCT and angiography through cardiac puncture. The cause of death was cardiac tam-ponade and right ventricular rupture due to the crush injury of chest in the traffic accident. In case 2, splenic rupture and intra-abdominal hemorrhage was found and caused by injury of left trunk by MSCT. The cause of death was hemorrhage and traumatic shock. Conclusion MSCT could observe skeletal in-jury, soft tissue injury, and hematologic disorder well. The combination use of MSCT and angiography through cardiac puncture provided assistance to the diagnosis of cardiovascular system injury.
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Objective T o observe the chem ical groups changing in rat kidney w ith regard to fatal hyper-therm ia by Fourier transform infrared m icrospectroscopy (FT IR-M SP ) and to provide a new m ethod to diagnose fatal hypertherm ia. Methods R ats w ere sacrificed by hypertherm ia, brainstem injury, m assive hem orrhage and asphyxiation and divided into groups. T he renal sam ples w ere dissected im m ediately af-ter death. T he data of infrared spectroscopy in glom erulus w ere m easured by FT IR-M SP. Results T he absorbances of 3 290, 3 070, 2 850, 1 540 and 1 396 cm -1 significantly increased (P<0.05),and the ratios of A1650/A3290 and A1650/A1540 significantly decreased (P<0.05) in group of hypertherm ia. Conclusion FTIR-M SP can analyze the changes of chem ical groups of kidney as an auxiliary diagnosis for discrim inating hyper-therm ia w ith other causes of death.
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A 45-year-old male car driver died in a traffic accident of four cars rear-end collision on the highway. He was found to have died after a respiratory and cardiac arrest at the scene. No sign of skin injuries was observed from the external inspection. The autopsy was not permitted by the family members because of the local culture. Multislice computed tomography (MSCT) was applied to the current case, showing dislocation of C3~4 cervical vertebrae with Ⅱ degree, C4 vertebral plate fractures, and spinal stenosis. Post-mortem MSCT confirmed the diagnosis as whiplash injuries. MSCT was verified to be effective in showing the severity of whiplash injuries, thus providing certain objective evidence for medicolegal expertise.
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Objective To build a high quality diagnosis system for schistosomiasis surveillance in the situation of low infec-tion in Jianglin County. Methods The network laboratory for schistosomiasis diagnosis was built according to the national crite-ria in Jianglin County in 2012. Results The network laboratory for schistosomiasis diagnosis was established successfully and the operation was quiet well. Conclusion The establishment and operation of the laboratory play an important role in the real-ization of schistosomiasis elimination.
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Multidrug resistance (MDR) plays a major obstacle to successful gastric cancer chemotherapy. The purpose of this study was to investigate the MDR reversal effect and mechanisms of hyperthermia in combination with neferine (Nef) in adriamycin (ADM) resistant human SGC7901/ADM gastric cancer cells. The MDR cells were heated at 42°C and 45°C for 30 min alone or combined with 10 μg/mL Nef. The cytotoxic effect of ADM was evaluated by MTT assay. Cellular plasma membrane lipid fluidity was detected by fluorescence polarization technique. Intracellular accumulation of ADM was monitored with high performance liquid chromatography. Mdr-1 mRNA, P-glycoprotein (P-gp), γH2AX expression and γH2AX foci formation were determined by real-time PCR, Western blot and immunocytochemical staining respectively. It was found that different heating methods induced different cytotoxic effects. Water submerged hyperthermia had the strongest cytotoxicity of ADM and Nef combined with hyperthermia had a synergistic cytotoxicity of ADM in the MDR cells. The water submerged hyperthermia increased the cell membrane fluidity. Both water submerged hyperthermia and Nef increased the intracellular accumulation of ADM. The water submerged hyperthermia and Nef down-regulated the expression of mdr-1 mRNA and P-gp. The water submerged hyperthermia could damage DNA and increase the γH2AX expression of SGC7901/ADM cells. The higher temperature was, the worse effect was. Our results show that combined treatment of hyperthermia with Nef can synergistically reverse MDR in human SGC7901/ADM gastric cancer cells.
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@#ObjectiveTo study the effects and molecular mechanisms of Shuanglong Pill on angiogenesis of ischemic myocardium. Methods63 Wistar rats were ligated left anterior descending branch to be made into models of acute myocardial infarction(AMI).All rats were randomly divided into large-dosage group of Shuanglong Pill(LDG),small-dosage group of Shuanglong Pill(SDG),The control of acute MI(MIC).Other 11 rats were set as normal control(NC).Rats in every group had been treated half for 2 weeks and half for 4 weeks. New vessels' quantity in ischemic myocardium of acute MI rats were detected by immunohistochemistry double steps assay;the expressions of vessel endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) on ischemic myocardium were detected by immunohistochemistry SP assay;the expressions of VEGF mRNA and bFGF mRNA on ischemic myocardium were detected by reverse transcription polymerase chain reaction(RT-PCR) assay.ResultsThere were more new capillaries on ischemic myocardium of LDG, SDG compared with that in MIC at every stage,moreover,there was significantly increase at the end of 4 weeks compared with that at 2 weeks.At the end of 2 weeks, the expression of VEGF in LDG were significantly elevated compared with that in MIC; and the expression of bFGF in LDG were significantly elevated compared with that in SDG and MIC.Whereas the expression of VEGF in LDG were significantly decreased at the end of 4 weeks compared with 2 weeks. There were significant differences of above comparison (P<0.05-0.01).At every stage the expression of VEGF mRNA in LDG, SDG were significantly elevated compared with that in MIC; but its expression was significantly decreased at the end of 4 weeks compared with that 2 weeks. The expression of bFGF mRNA was significantly elevated in LDC at the end of 2 weeks compared with that in other groups.ConclusionsShuanglong Pill could promote angiogenesis in ischemic myocardium,application of large dose of Shuanglong Pill could significantly upregulate the expression of VEGF, bFGF, VEGF mRNA and bFGF mRNA.
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Objective IFN-? is a pleiotropic cytokine with potent immunomodulatory effects and antiviral activity. To study the mechanism of IFN-? clearing duck hepatitis B virus (DHBV) in ducks, it is essential to establish a method to quantify expression of DuIFN-? in immune response. In the present study,a semi-quantitative competitive RT-PCR was developed to quantify expression of duck IFN-?(DuIFN-?) mRNA by PBMCs. Methods Based on ?-actin consensus sequence, fishing the ?-actin gene as house keeping gene from duck PBMC by RT-PCR. A competitive internal control was constructed and the competitive RT-PCR system could be used to quantify the transcription of DuIFN-? mRNA. Results After duck PBMCs were stimulated in vitro with PHA, the peak of DuIFN-? expression was at 24-36h. Then RT-PCR method was applied to detect DuIFN-? mRNA transcription by PBMCs from DHBV infected ducks immunized with DuIFN-? plasmid plus DNA vaccine or DNA vaccine alone. Results showed that expression of DuIFN-? in ducks co-immunized with DuIFN-? plasmid were higher than other groups immunized without DuIFN-? plasmid as adjuvant. Conclusions The results indicated that DuIFN-? gene could be a useful adjuvant to develop vaccines. The semi-quantitation of DuIFN-? mRNA by competitive RT-PCR provides the basis for future study of the mechanism of IFN-? in duck hepatitis B virus persistent infection.