ObjectiveTo facilitate the establishment of fully automatic clinical diagnostic nucleic acid assay,a study was carried out for developing a fast and convenient method of extracting highly purified viral nucleic acid from human blood with magnetic beads.Method Five KHB (Shanghai Kehua Biological Engineering Company Limited) internal strong positive HBV serum samples were used in the process,for the method development,screening and selecting the magnetic beads based on magnetism,suspension power and extraction effects of HBV DNA,the optimization of the selected key components of extraction reagents were performed,which including the quantity of magnetic beads( 1 - 1.75 μl/ml),the buffer pH of lysis (4 -9),washing conditions with or without alcohol,elution for proliferation with or without magnetic beads,and etc.The evaluation for the extraction method was carried out including linearity,precision,sensitivity,specificity,anti-interference and resistance to contamination.The magnetic beads method was compared with domestic mature boiling method in a quantitative test with 100 HBV serum samples,the magnetic beads method was also compared with the interntional Gold standard Roche COBAS Tagman test method ( Roche method) in a quantitative test with 50 HBV serum samples and the results were statistically compared for their linear regression.ResultsOptimization experiment result showed superparamagnetic beads A was the best among the candidates ( C,B,D) in terms of their physical properties and extraction effects of DNA.The optimum condition of the assay were as follow:magnetic beads best used at 1.25 μl/ml,buffer pH 7 - 8 of lysis,ethanol was eliminated from washing buffer,proliferation can be carried out with magnetic beads.The parallel comparison experiments with five KHB internal strong positive HBV serum sample and gradient diluted serum sample between beads method and boiling method were carried out.The results showed that the linear range of beads method was 4 × 101 -4 × 108 IU/ml and that of boiling method was 4 × 102 -4 × 108 IU/ml.The sensitivity for beads method was 30 IU/ml and that of boiling method was 100 IU/ml.The precision CV of beads method was 6.0％ and boiling method was 11.9％.The false positive rate was 0 for beads method vs 3.1％ for boiling method.The anti-interference ability experiment showed that when sample contained sodium heparin concentration exceed 62.5 U/ml, there was significant drop of HBV DNA detection with boiling method and no HBV DNA was detected when sodium heparin concentration in the sample reached 1000 U/ml.However,sample sodium heparin concentration had no effect on HBV DNA detection with beads method.The comparison study between beads method and boiling method was carried out by use of 100 clinical HBV positive serum samples.The result showed that the sensitivity for beads method was 86％ vs boiling method 74％.The correlation between the two method was R2 =0.948,P ＜ 0.05.The comparison study between beads method and Roche method was carried use of 50 clinical HBV DNA positive serum samples and the result showed that sensitivity for beads method was 95％ (49/50) vs Roche method was 100％ (50/50).The correlation was R2 =0.963,P ＜ 0.05.ConclusionsThe established method for automatic nucleic acid extraction with magnetic bead displayed characteristic of high yield nucleic acid extraction,broader testing range,accurate quantification and convenient in operation.This can be one choice for nucleic acid extraction and applicable for clinical automatic diagnostic system.( Chin J Lab Med,2012,35:843 -850 )

ObjectiveTo assess the efficacy of herpes virus entry mediator (HVEM) gene modifled dendritic cells (DCs) in protecting against myosin induced myocarditis,and to investigate the involving mechanism.MethodsWe treated experimental autoimmune myocarditis (EAM) mice with myosin-pulsed DCs which were transfected with HVEM-expressing adenovirus (Ad-HVEM) or control vectors,then evaluated myocarditis,plasm cTn [ and autoantibody by histopathology,fluoroimmunoassay,and ELISA,respectively.ResultsWe found that DCs transfected with Ad-HVEM (DC-Ad-HVEM) could protect against EAM.Further study showed DC-Ad-HVEM could produce regulatory cytokine IL-10,and IL-10 promoted the production of a key regulatory T cell subset which is important in peripheral tolerance.The T cells mediated protection against EAM.ConclusionThis study suggest that myosin-DC-Ad-HVEM cell gene therapy is a safe and effective way for inhibiting the development of EAM,and the signal net mediated by HVEM plays different roles in different cells.

Objective To explore the relationship between 5-minute Apgar score and heart rate variability (HRV) in severely asphyxiated neonates. Methods A total of 103 severely asphyxiated neonates with 1-minute Apgar score of 0 to 3 points were selected. They were divided into Group A (>7 points, n=50) and Group B (≤7 points, n=53) based on 5-minute Apgar score. Meanwhile, 40 full-term neonates with 1-and 5-minute Apgar score greater than 7 points were selected as control group. 24-hour dynamic electrocardiogram was performed and HRV was analyzed on the third day after birth in three groups. Results PNN50, rMSSD, SDSD were decreased and SDNN, SDANN were increased in group B as compared with group A and control group (P<0.05). No signiifcant difference of HRV was observed between group A and control group (P>0.05). Conclusions Neonatal asphyxia can cause damage to autonomic nervous system. 5-minute Apgar score and HRV can be joint-ly used as a non-invasive index in autonomic nervous damage and its prognosis in asphyxiated newborns.

Objective To evaluate the clinical value of color Doppler ultrasound (CDUS) and computed tomography angiography (CTA) in diagnosis of Budd-Chiari syndrome(BCS).Methods The CDUS and CTA data about 33 BCS patients diagnosed by digital subtraction angiography(DSA) and treated by interventional therapy were retrospectively analyzed.All of 33 patients with BCS were examined with CDUS,26 cases of then received CTA.The diagnostic value of BCS between two methods were compared.Results The diagnostic accordance rate of CDUS and CTA for BCS were 90.9％ and 96.2％ respectively,which had no significant statistical difference(P ＞ 0.05).The two methods could not only comprehensively systematically reflect the morphologic changes of liver and spleen,the stenosis and blood flow state of the lesion regions,but also clearly display the intrahepatic/extrahepatic collateral vessels trend in the abdomen.One week after interventional therapy,the CUDS results showed blood vessels pristine and stable hemodynamics.Conclusion CDUS is the preferred imaging in the diagnosis and postoperative re-examination of BCS.While,CTA can provide many important parameters for preoperative assessment and operation way.

Objective To evaluate the safety and efficacy of laparoscopy-assisted total gastrectomy (LATG) for gastric cancer by systematic review and meta-analysis.Methods The literature database before February,2012 was extensively searched to retrieve the comparative studies of LATG and open total gastrectomy(OTG) with a relevance of study goal.The inclusion and exclusion criteria were formulated.Aftera quality evaluation,the data was extracted.The Cochrane collaboration RevMan 5.1 version software was used for meta-analysis.Results There are ten studies meeting the inclusion criteria for meta-analysis.The total sample size of these studies was 942 cases.Compared to OTG,LATG experienced longer operation time [weighted mean difference(WMD) =41.12 min,95％ confidence interval(CI):20.62-61.63,P ＜0.01)],less blood loss(WMD =-198.36 ml,95％ CI:-300.94--95.78,P ＜ 0.01),earlier time to flatus(WMD =-0.80 days,95％ CI:-1.17--0.42,P＜0.01),shorter hospital stay(WMD =-4.02days,95％ CI:-6.03--2.01,P ＜ 0.01) and decrease in overall complications [relative risk (RR) =0.55,95％ CI:0.40-0.76,P ＜ 0.01)].The number of dissected lymph nodes,proximal or distal resection margin and mortality were similar between the two groups.Conclusions Laparoscopy-assisted total gastrectomy is a safe and feasible procedure with less blood loss,less overall complications and quick recovery.

Objective To investigate clinical efficacy of Easy First strategy in laparoscopic pancreaticoduodenectomy (PD) for borderline resectable pancreatic cancer.Methods The clinical data of 9 patients with borderline resectable pancreatic cancer who were admitted to the Sir Run Run Shaw Hospital of Zhejiang University (8 patients) and Zhejiang Provincial People's Hospital (1 patient) from June 2013 to March 2015 were retrospectively analyzed.Nine patients underwent laparoscopic pancreaticoduodenectomy based on the Easy First strategy (to sequentially dissect and amputate jejunum,stomach,hepatoduodenal ligament,common bile duct,main portal vein,head of pancreas,second segment and third segment of the duodenum,uncinate process and neck of pancreas).The operation time,volume of intraoperative blood loss,postoperative pathological examination,complications and duration of hospital stay were recorded.Patients were followed up once every 3 months by outpatient examination and telephone interview up to June 2015.Results Of 9 patients,4 received totally laparoscopic PD (2 received partial resection and repair of portal venous wall),1 received laparoscopic assisted resection and digestive tract reconstruction,and 4 received laparoscopic transection of jejunum,bile tract and stomach and conversion to open surgery for resection and digestive tract reconstruction (1 with severe adhesion between tumor and portal vein,3 with bleeding due to dissection of neck and unciform process of pancreas).The operation time and volume of intraoperative blood loss in all patients and in 4 patients with totally laparoscopic PD were (404 ± 49) minutes and (456 ± 348) mL,(395 ± 61) minutes and (188 ± 25) mL,respectively.Of 9 patients,5 with postoperative complications were cured without perioperative death,including 2 with Grade B pancreatic fistula,1 with biliary leakage,1 receiving reoperation due to gastric stump bleeding at postoperative day 7 and 1 with abdominal infection.The extubation time of right drainage tube and left drainage tube was (9 ± 5) days and (11 ± 4) days,respectively.The duration of hospital stay was (24 ± 10)days.All patients were diagnosed as with pancreatic cancer by pathological examinations with the tumor diameter of (3.2 ± 0.8) cm.The number of harvested lymph nodes in all patients and in 4 patients with totally laparoscopic PD were 16.8 (range,6.0-25.0) and 19.8 (range,15.0-25.0).All the patients were followed up for mean time of 12 months (range,4-24 months),including 1 death at postoperative month 3,1 with tumor survival of 20 months and others with tumor-free survival.The postoperative survival time of 4 patients was more than 18 months at the end of follow-up.Conclusion Easy First strategy in laparoscopic PD is safe,feasible and practical for borderline resectable pancreatic cancer.

Objective To compare and analyze the tilt and decentration of the intraocular lens in patients receiving four-point and two-point suspension fixation,as well as their relationship with visual prognosis.Methods A total of 80 patients(80 eyes)who underwent intraocular lens suspension fixation at the Ophthalmology Department of Baoding No.1 Central Hospital from June 2021 to April 2022 were selected as the subjects.These patients were randomly divided into the experimental group(41 patients,41 eyes,underwent four-point suspension fixation)and the control group(39 patients,39 eyes,underwent traditional two-point suspension fixation).They were followed up for at least 6 months after surgery to re-cord their uncorrected visual acuity(UCVA)and best corrected visual acuity(BCVA)before surgery and at the last follow-up.The tilt angle and decentration distance of the intraocular lens of patients in the two groups were measured after surger-y by a panoramic ultrasound biomicroscope.The preoperative and last follow-up UCVA and BCVA of patients in the two groups,as well as tilt angle and decentration distance of the intraocular lens after surgery,were compared,and the corre-lation between tilt angle,decentration distance and postoperative UCVA,BCVA was analyzed by Person correlation analy-sis.Results The UCVA and BCVA at the last follow-up in the experimental group and control group were better than those before surgery(all P＜0.05).The difference in postoperative UCVA between the experimental group and the control group was statistically significant(t=-6.20,P=0.00),and the experimental group had better postoperative UCVA than the control group.There was no statistically significant difference in postoperative BCVA between the experimental group and the control group(t=-1.43,P=0.16).The postoperative horizontal and vertical tilt angles of the intraocular lens in the experimental group were 0.70°±0.24° and 0.60°±0.16°,respectively;while those in the control group were 2.66°± 1.40° and 3.76°±0.67°,respectively.The differences between the two groups were statistically significant(t=-8.51 and-29.42,P=0.00 and 0.00).The postoperative horizontal and vertical decentration distances of the intraocular lens in the experimental group were(0.24±0.10)mm and(0.25±0.10)mm,respectively,while those in the control group were(0.85±0.77)mm and(2.14±0.50)mm,respectively.The differences between the two groups were statistically signifi-cant(t=-4.82 and-21.68,P=0.00 and 0.00).In the experimental group,neither the horizontal and vertical tilt angles of intraocular lenses nor the horizontal and vertical decentration distances were correlated with postoperative UCVA and BCVA(all P＞0.05).In the control group,the horizontal tilt angle of intraocular lenses was positively correlated with post-operative UCVA and BCVA(both P＜0.05),while the vertical tilt angle was not correlated with postoperative UCVA and BCVA(both P＞0.05);the horizontal decentration distance was positively correlated with postoperative UCVA and BCVA(both P＜0.05),but the vertical decentration distance was not correlated with postoperative UCVA and BCVA(both P＞0.05).Conclusion Both four-point suspension fixation and traditional two-point suspension fixation can effectively im-prove postoperative vision of patients,while the tilt and decentration of the intraocular lens are smaller after four-point sus-pension fixation.

Objective To analyze the status of coronary collateral circulation(CCC)in patients with acute myocardial infarction(AMI)and explore its relationship with thrombus burden.Meth-ods Convenient sampling was used to select 161 AMI patients diagnosed and treated in Tongling People's Hospital from May 2022 to March 2024.Their CCC status was assessed by Rentrop clas-sification,and according to the results,they were divided into well-(grade 2～3,84 cases)and poorly-developed CCC groups(grade 0～1,77 cases).The general clinical data,cardiac function in-dicators and thrombus burden were compared between the two groups.Multivariate logistic re-gression was employed to identify the independent influencing factors for CCC status in AMI pa-tients,and ROC curve was plotted to analyze the relationship between CCC status and thrombus burden.Results In the 161 AMI patients,the incidence of adverse CCC was 47.83％(77/161).The poorly-developed CCC group had significantly lower Gensini score,and higher white blood cell count than the well-developed CCC group(P＜0.01);There is a statistically significant differ-ence in proportion of infarct related blood vessels between two groups(P＜0.01).The rate of high thrombus burden in the poorly-developed CCC group was obviously higher than that in the well-developed group(79.22％vs 26.19％,P＜0.01).Multivariate logistic regression analysis showed that infarction-related vessels(left anterior descending coronary artery),white blood cell count and thrombus burden were independent risk factors for poor development of CCC,while Gensini score was independent protective factors(OR=2.051,95％CI:1.071-3.928;OR=1.647,95％CI:1.353-2.006;OR=6.418,95％CI:2.654-15.519;OR=0.973,95％CI:0.948-0.999).ROC curve analysis indicated that the AUC value of thrombus burden was 0.752(95％CI:0.678-0.817).Conclusion It is quite common of poorly developed CCC in AMI patients,which is associ-ated with the increase of thrombus burden.

Objective:To investigate the efficacy and safety of daratumumab in the treatment of systemic light chain amyloidosis.Methods:The clinical data of 24 patients with systemic light chain amyloidosis who received daratumumab-based regimens in Sichuan Provincial People's Hospital from January 2020 to November 2022 were retrospectively analyzed. The treatment process of patients was summarized and the therapeutic efficacy was evaluated. Kaplan-Meier method was used to make survival analysis and the adverse reactions were analyzed.Results:All 24 patients included 2 cases (8.33%) of Mayo 2004 stageⅠ, 2 cases (8.33%) of Mayo 2004 stage Ⅱ and 20 cases (83.33%) of Mayo 2004 stage Ⅲ. All patients were treated with daratumumab-based regimen, and 17 patients had evaluable efficacy. In the chemotherapy regimens, 15 patients received DVd (daratumumab + bortezomib + dexamethasone) regimen, 7 patients received DVCd (daratumumab + bortezomib + cyclophosphamide + dexamethasone) regimen, 1 patient received DRd (daratumumab + lenalidomide + dexamethasone) regimen, and 1 patient received DTd (daratumumab +thalidomide + dexamethasone) regimen. After 1 course of daratumumab-based regimens in 17 cases with evaluable efficacy, the strict complete remission (sCR) rate was 41.18% (7/17), the overall response rate (ORR) was 88.24% (15/17). Among 17 patients who received daratumumab-based chemotherapy regimen as the first-line treatment, sCR rate of 11 cases with evaluable efficacy was 36.36% (4/11) after 1 course of treatment ORR was 90.90% (10/11). Among 5 relapsed/refractory patients, sCR rate of 4 cases with evaluable efficacy was 50.00% (2/4) after 1 course of treatment; ORR was 75.00% (3/4). Among 24 patients, renal involvement was found in 17 patients at the initial diagnosis. After 1 course of daratumumab-based chemotherapy regimen, ORR of 7 cases with evaluable efficacy was 85.71% (6/7), among which 42.86% (3/7) patients with renal involvement had an assessed renal response of very good partial remission (VGPR) or above. At the initial diagnosis, 19 cases had cardiac involvement; ORR of 14 cases with evaluable efficacy was 85.71% (12/14), among which 42.86% (6/14) patients had cardiac response to VGPR or above. After daratumumab-based chemotherapy regimen, the main adverse reactions were infusion-related adverse reactions, myelosuppression and infection, all of which were tolerated by the patients. The median follow-up time of 24 patients was 7.0 months (0.5- 16.5 months), the median progression-free survival time was 7.0 months (0.5-16.5 months) and the median overall survival time was 7.0 months (0.5-35.0 months).Conclusions:Daratumumab-based chemotherapy regimen has good efficacy and safety in the treatment of systemic light chain amyloidosis.

Objective To investigate the clinical effect of free pre-expanded scapular skin flap for repairing of large size facial and cervical scar .Methods 15 patients suffering from post-burn facial and cervical scar contractures were treated in the Department of Plastic Surgery of Second Affiliated Hospital of Kunming Medical University.The reconstruction was performed in two operative stages .First, the skin incision were made paralleling with the descending branches of the circumflex scapular artery near posterior axillary line and the scapular skin flap was elevated .A 300 to 400 ml kidney -shaped expander was implanted under scapular region skin .The expansion began 1 week post-operatively.After the expander was fully expanded and could supply sufficient flap , the facial and cervical scar was resected and the contracture was released .The pre-expanded scapular flap was harvested and transferred to repair the defects.Then the facial vascular anastomosis with circumflex scapular vascular was performed .The expander was removed and the wound was closed directly .Results The expansion time ranged from 2 to 4 months with the average time of 2.8 months.The flap size ranged from 14 cm ×7 cm to 25 cm ×14 cm. All flaps survived post-operatively and wounds at donor sites healed primarily .The face and neck have good appearance .Conclusions Pre-expanded scapular skin flap is suitable for repairing of larger face and neck scar with good color and thickness match .Expanded skin flap can provide large size flap , leaving less morbidity at the donor sites .