Three primer were designed based on the consevered area of the genetic of the ATCC VR-2332 strain and LV strain. And the nest RT-PCR of testing porcine reproductive and respiratory syndrome virus were developed. The nest RT-PCR against ATCC VR-2332 strain, LV strain and B13 strain were done by this method.The DNA fragment were obtained specially from the three strains isolated from different region. The size were 430bp (430bp) , 410bp (413bp) and 410 bp (413bp) separately. But the DNA fragment were not obtained from HCV, PPV and PRV. Its sensitivity was 10-2 TCID50. It's sensitivity increased 10000 times than one step RT-PCR. It should make the method of testing porcine reproductive and respiratory syndrome virus more sensitive, fast and accurate.

A nest RT-PCR/restriction test has been developed in order to distinguish the lapinised vaccine strain from field isolates of classical swine fever virus. The restriction enzyme cut sites mapping of the major coding sequence of E2 gene lapinised vaccine strain and ShiMen strain of classical swine fever virus have been compared. Ten and sixteen unique restriction markers have been found in the lapinised vaccine strain and ShiMen strain. The restriction enzyme cut sites mapping of the twenty six unique restriction marker in the major coding sequence of E2 gene of 17 classical swine fever field isolates have been analyzed. Only 3 sites (HgaI、Hin8I及Hsp92I) are present in the lapinised vaccine strain sequence. Two pans of nested primers and a criteria of analysis have been designed for HgaI restriction marker site. The tests have been conducted first on the lapinised vaccine strain and ShiMen strain of classical swine fever virus resulting in predicted restrection patterns. Finally, the tests have been applied to 5 field isolates of different gene group analyzed by phylogenetic study. The result showed that only HCLV strain gene can be cut to 2 fragment by Hgal , and ShiMen strain and 5 field isolates cant be cut At the same time the sensitivity and specificity of nest RT-PCR have been tested. The sensitivity is 0. 2MLD. The specific fragment of BDV and BVDV were not obtained by the nest RT-PCR. These results showed that the development of the nest RT-PCR/restriction tests is very important for the control and perish of classical swine fever in china.

In this study, we evaluated the difference ot biological characteristics in the MERS-CoV infected mice model in prior to transduction with different dosage of human DPP4. Firstly, we transduced different dosage of DPP4 (high or low) into mice, and then challenged them with MERS-CoV in order to establish the model. After establishment of mice model, we observed the clinical signs of disease, virus replication, immunopathogenesis and antibody response. The results indicated that the infected mice showed typical pneumonia, virus replication, histological lesions, and neutralizing antibody production. Moreover, the high dosage group was superior to the low dosage group. Fourteen days after infection, the specific antibody to virus structural protein and neutralizing antibody were analyzed, the high dosage group induced higher level antibody. In summary, the MERS-CoV infected mice model were established prior transduction with DPP4, and the level of DPP4 influenced the clinical signs of disease, virus replication and antibody response in this model.
Animals ; Coronavirus Infections ; enzymology ; genetics ; pathology ; virology ; Dipeptidyl Peptidase 4 ; genetics ; metabolism ; Disease Models, Animal ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Middle East Respiratory Syndrome Coronavirus ; genetics ; physiology

Objective To study the prevalence of esophageal cancer and various lesions of esophagus in high risk areas through a screening program for early diagnosis and treatment.Methods Random cluster sampling method was used to select some portions of a natural village as screening object in the high risk areas of esophageal cancer,from 2006 to 2011.Endoscope iodine staining and index biopsy screening methods were used on people with high risk and followed by pathological exams for confirmation.Results The detection rates regarding mild esophageal hyperplasia,moderate and severe esophageal hyperplasia were 5.33％ (803/15 065),1.28％( 193/15 065 ),0.68％( 102/15 065 ) respectively while the detection rates on carcinoma in situ,intramucosal carcinoma and invasive cancer were 0.15％(22/15 065),0.06％(9/15 065),0.29％(43/15 065)respectively.The detection rate in male esophageal hyperplasia was higher than in female.People younger than 65 years old,the detection rates on mild,moderate or severe esophageal hyperplasia and invasive cancer showed an increase with age,with the 60- year-olds group reaching the highest.The detection rates on the above said diseases were 7.72％( 198/2565 ),2.07％(53/2565),1.29％( 33/2565 ),0.51％ ( 13/2565 ) respectively.The detection rates on mild,moderate or severe esophageal hyperplasia varied in different years and with statistically significant differences (P＜0.001) but did not show any obvious trend of changing.Geographical distribution of mild esophageal hyperplasia,moderate esophageal hyperplasia,severe esophageal hyperplasia also significantly varied in different villages (P＜0.001).The highest detection rate in the mountainous villages was seen the highest while the detection rate of village from hilly areas was the lowest.Conclusion There were considerable numbers of patients with precancerous lesions in the general population from the high risk areas.The detection rate of esophageal cancer in the mountain residents was higher than the rate in the hilly areas.Men and the elderly were the key populations calling for esophageal cancer prevention programs to be carried out.

Objective To analyze the high risk factors of obstetric brachial plexus palsy(OBPP),and to explore how to evaluate the relationship between fault medical behavior and OBPP in the process of medical damage forensic identification.Methods A retrospective analysis was carried out on 25 cases of medical damage liability disputes related to OBPP from 2017 to 2021 in Beijing Fayuan Judicial Science Evidence Appraisal Center.The shortcomings of hospitals in birth weight assessment,delivery mode selection,labor process observation and shoulder dystocia management,and the causal relation-ship between them and the damage consequences of the children were summarized.Results Fault medi-cal behavior was assessed as the primary cause in 2 cases,equal cause in 10 cases,secondary cause in 8 cases,minor cause in 1 case,no causal relationship in 1 case,and unclear causal force in 3 cases.Conclusion In the process of forensic identification of OBPP,whether medical behaviors fulfill diagno-sis and treatment obligations should be objectively analyzed from the aspects of prenatal evaluation,de-livery mode notification,standardized use of oxytocin,standard operation of shoulder dystocia,etc.Meanwhile,it is necessary to fully consider the objective risk of different risk factors and the diffi-culty of injury prevention,and comprehensively evaluate the causal force of fault medical behavior in the damage consequences.

OBJECTIVETo evaluate the value of magnetic resonance dynamic contrast-enhanced (MR-DCE) and magnetic resonance diffusion-weighted imaging (MR-DWI) in the differentiation of benign and malignant musculoskeletal tumors.

METHODSSixty-three patients with pathologically confirmed musculoskeletal tumors were examined with MR-DCE and MR-DWI. Using single shot spin echo planar imaging sequence and different b values of 400, 600, 800 and 1000 s/mm(2), we obtained the apparent diffusion coefficient (ADC) of the lesions. ADC values were measured before and after MR-DCE, with a b value of 600 s/mm(2). The 3D fast acquired multiple phase enhanced fast spoiled gradient recalled echo sequence was obtained for multi-slice of the entire lesion. The time-signal intensity curve (TIC), dynamic contrast-enhanced parameters, maximum slope of increase (MSI), positive enhancement integral, signal enhancement ratio, and time to peak (T(peak)) were also recorded.

RESULTSADC showed no significant difference between benign and malignant tumors when the b value was 400, 600, 800, or 1000 s/mm(2), and it was not significantly different between benign and malignant tumors in both pre-MR-DCE and post-MR-DCE with b value of 600 s/mm(2). TIC were classified into four types type1 showed rapid progression and gradual drainage; type2 showed rapid progression but had no or slight progression; type 3 showed gradual progression; and type 4 had no or slight progression. Most lesions of type1 or type2 were malignant, whereas most lesions of type 3 or type 4 were benign. When using type1 and type 2 as the standards of malignancy, the diagnostic sensitivity and specificity was 87.23% and 50.00%, respectively. The types of TIC showed significant difference between benign and malignant musculoskeletal tumors(χ(2)=17.009,P=0.001). When using MSI 366.62 ± 174.84 as the standard of malignancy, the diagnostic sensitivity and specificity was 86.78% and 78.67%, respectively. When using T(peak)≤70s as the standard of malignancy, the diagnostic sensitivity and specificity was 82.89%and 85.78%, respectively. Positive enhancement integral and signal enhancement ratio showed no significant difference between benign and malignant musculoskeletal tumors.

CONCLUSIONSTIC, MSI and T(peak) of MR-DCE are valuable in differentiating benign from malignant musculoskeletal tumors. T(peak) has the highest diagnostic specificity, and TIC has the highest diagnostic sensitivity. The mean ADC value are no significant difference between benign and malignant tumors.

Adolescent ; Adult ; Aged ; Bone Neoplasms ; diagnosis ; Child ; Diagnosis, Differential ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Muscle Neoplasms ; diagnosis ; Young Adult

Objective:To investigate the feasibility and applicability of laparoscopic totally extraperitoneal (TEP) inguinal hernia repair by using one-point-suturing fixed soft mesh.Methods:90 male patients were retrospectively analyzed from Jan.2013 to Mar.2015,all of them were diagnosed with unilateral inguinal hernia and treated with TEP surgery.Among them,there were 55 patients with free of air bag and free of fixed TEP (free fixed group) and 35 patients with free of air bag and fixed by one-point-suturing TEP (one-point-suturing fixed group),all of patients were used by soft mesh.They were analyzed about their differences of postoperative pain,gastrointestinal function recovery time,the postoperative feeding time,the bedside movement time,the hospitalization time,and the incidence of postoperative complications.Results:Compared with the free fixed group,the operation time of the one-point-suturing fixed group was significantly shorter,the bedside movement time earlier,gastrointestinal function recovery time earlier,the postoperative feeding time earlier(P＜0.05),but the difference were not statistically significant in intraoperative blood loss,the postoperative pain,the hospitalization time,the incidence of postoperative seroma (P＞0.05),and therewere no recurrence in both groups.Conclusion:Compared with the free fixed TEP,the one-point-suturing fixed TEP has significantly shorter operation time,reduce the operation difficulty,and make the postoperative recovery faster,so it is safe,feasible and appropriate to application.

Objective To investigate the lung protection of remifentanil in rats with acute severe hemorrhagic shock (ASHS).Methods A total of 50 healthy adult SD rats were selected and randomly divided into model group,sham-operation group and low,middle,high dose test groups,10 rats in each group.Model group were ASHS model rats.Other groups were intravenous given injection with loading-dose of remifentanil 1 μg · kg-1,then low,middle and high dose test groups were intravenous infusion with 0.2,0.6,1.2 μg · kg-1 · min-1 remifentanil.The lung hilus in model group and sham-operation group were dissociate without blockage,while model group and sham-operation group were infused with 0.9％ NaC1 as the same dose of high-dose test group.The injuries of lung tissues in rats were judged by pathological examination.The wet to dry weight ratio (W/D) of lung tissue,malondialdehyde (MDA),activity of superoxide dismutase (SOD),content of nitrogen oxide (NO) were detected in each group.Results The W/D,MDA,SOD,NO in model group were 2.97 ± 0.17,(3.45 ± 1.48) nmol · mg-1 prot,(11.73 ± 1.78)U · mg-1 prot,(0.13 ±0.03)μmol · mg-1 prot,had no significant difference with those in low dose test group,which were 2.71 ± 0.12,(2.90 ± 1.03) nmol · mg-1 prot,(14.44 ± 1.55) U · mg-1 prot,(0.16 ± 0.05) μmol · mg-1 prot (all P ＞ 0.05).The W/D,MDA,SOD,NO in middle-dose test group were 2.33 ±0.16,(2.29 ±0.72) nmol · mg-1 prot,(15.59 ± 1.80)U · mg-1prot,(0.19 ±0.05) μmol· mg-1prot,and those in high-dose test group were 2.08 ± 0.07,(1.81 ± 0.64)nmol · mg-1 prot,(17.47 ± 1.10)U · mg-1prot,(0.26 ±0.08) μmol · mg-1prot,these indexes in middle-dose and high-dose test groups had significant difference with model group (all P ＜ 0.05).Conclusion Remifentanil can inhibit lipid peroxidation,reduce injury of lung tissue,so as to protect the lung of ASHS rats.

AIM:To evaluate the efficacy and safety of "one-stitch anastomosis through the skin" repair of canalicular laceration.METHODS:The data of 32 cases (32 eyes) of canalicular laceration who underwent repair of lacerated canaliculi with one-stitch anastomosis through the skin were retrospectively reviewed, inferior canalicular laceration in 29 patients,superior canalicular laceration in 1 patient, 2 cases involving both the inferior and superior canalicular laceration. All the operations were performed under surgical microscope, 5-0 silk sutures were used and silicone tube of 0.8mm diameter was employed in intubation. The stents were left in place for 3 months postoperatively and then removed. The follow-up period was 1 to 36 months.RESULTS: In 32 patients, 28 (88%)patients were cured entirely, 3 (9%)patients were meliorated, and 1 (3%)patient had no effects. A total of 29 patients complied with scheduled follow up 1-36 months (average 12 months) after stent removal, and 3 patients were lost in follow-up. All the patients had got good recovery of eyelid laceration with no traumatic deformity in eyelid and canthus.CONCLUSION: In "one-stitch anastomosis through the skin"repair of canalicular laceration, the cut ends could be anastomosed directly,for there was no suture remained in the wound permanently, so there was no suture-related granuloma which might cause obstruction or stenosis of canaliculi. It was simple, economical ,effective and safe.

Objective To develop a method for the determination of total cholesterol in serum by isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS).Methods Serum samples were supplemented by addition of [3,4-~(13)C_2]-cholesterol,hydrolyzed with alcoholic sodium hydroxide and oxidized into cholest-4-ene-3,6-dione by chromic acid.The oxidation products were analyzed by LC/MS/MS using atmospheric pressure chemical ionization (APCI) source and detection modes of multiple reaction monitoring (MRM) and single ion recording (SIR).Signals (peak areas) of the internal standard were corrected for the contributions of cholesterol and the signal ratios of cholesterol to internal standard for the calibrations were linearly regressed against cholesterol concentrations.The resulted regression equation was used for the calculation of serum cholesterol concentrations.Results The correlation coefficients between the peak area ratios and cholesterol concentrations were 0.999 9 and higher.Under MRM mode,the average within-run CV of the results obtained on 3 serum samples was 0.95% (ranged from 0.92% to 0.99%) and the total CVwas 0.86% (0.82% to 0.89%),and under SIR mode,the within-run CV was 0.64% (from 0.54% to 0.77%) and the total CVwas O.69% (0.62% to 0.81%),respectively. Results on certified reference materials (SRM 1951 a Level Ⅰ and Level Ⅱ;GBW 09145 and GBW 09147) showed an average bias of 0.23% (0.14% to 1.00%) under MRM mode,and 0.24% (0.07% to 1.27%) under SIR mode.Conclusions An ID-LC/MS/MS method for serum cholesterol has been developed.It is specific and precise and may be used as a candidate reference method.