s:To resolve the problem of the accuracy and standardization of STR PCR typing in forensic science practice,we have designed a new method to produce standard D12S375 allelic ladder.Seven different PCR amplified D12S375 allelic fragments were isolated from the gel,eluted into the distilled water and reamplified by PCR.The purified allelic fragments were then blunt end subcloned individually into the pUC plasmid vectors and transfected into competent E.coli DH5? TM cells.The sequencing results confirmed that the size and the constructure of the inserts were correct.The recombinant plasmids DNA with 7 inserts were then used as templates for reamplification to generate D12S375 standard ladder, with which the genetic polymorphisms of D12S375 locus in Chinese Han population in Chengdu,Hui population in Gansu and Wei population in Xinjiang were studied.

Objective To explore the efficacy of endoscopic therapy for common bile duct stones in the elderly. Methods From July 2003 to June 2006,421 elderly patients with common bile duct stones were treated in the Danyang People's Hospital(112 cases) and the Second Affiliated Hospital of Nanjing Medical University(309 cases).The clinical data of the patients were retrospectively analyzed Results ERCP was completed in all the 421 patients,of which 27 was converted to a pre-cut sphincterotomy because of difficulties in the deep cannulation of the bile duct.The diagnosis of common duct stones was confirmed during the operation in all the cases,and 46 of them were found complicated with cholecystolithiasis.Of the patients,266 patients had one stone,107 had two,and 48 had three or more.The calculi were removed using basket directly in 283 patients,balloon catheter in 122,lithotrity in 10,and plastic stent placement without calculus removal in 6.The stones were removed completed at the first operation in 392 cases,and the other 23 cases received a second operation after endoscopic nasobiliary drainage.Errhysis occurred in 20 patients during the pre-cut sphincterotomy,and was healed by locally sprinkling or injecting noradrenaline(1:10 000),or electric coagulation.Six patients had hemorrhage from the incision on the duodenal papilla 48 hours after the operation,and were cured by using submucous injection of noradrenaline(1:10 000,4 cases) or titanium clipper(2 cases).34 patients had a transient increase of serum amylase without abdominal pain;10 patients developed mild pancreatitis and then was cured by fasting,inhibiting pancreatic secretion and pancreatic enzyme activities,anti-inflammation therapy,and rehydration;4 patients developed cholangitis,and was cured by anti-inflammation therapy.All the patients were discharged without complications.342 of the patients were followed up for 6-12 months,4(1.2%) of them had recurrence of stones and was treated with endoscopy,3(0.9%) had cholangitis and received anti-inflammation therapy.In the 6 patients with plastic stent,the stones were dissolved in 2 of them in half a year.Conclusions Endoscopic therapy is safe and efficient for common bile duct stones in the elderly and is associated with a low rate of complications.

Objective To analyze the importance of procalcitonin (PCT) and high-sensitivity Creactive protein (hsCRP) in assessing the severity of pneumonia and sepsis patients as well as prognostic evaluation.Methods A total of 77 patients with pneumonia complicated with sepsis were randomly (random number) selected from May 2013 to May 2016 in our hospital and 50 patients with simple pneumonia were enrolled as control group.The sepsis pneumonia patients were divided into three groups,namely sepsis group,severe sepsis group and septic shock group.The sepsis patient were further divided into survival group and death group according to the death of patient within 2 weeks.Statistics was employed to study the roles of PCT and hsCRP in evaluating the severity of pneumonia and sepsis patients as well as prognostic evaluation.Results Compared with control group,the levels of PCT and hsCRP were higher in patients of sepsis groups (P ＜ 0.05).The levels of PCT and hsCRP were gradually increased as the severity of the patient getting worse (P ＜ 0.05).The levels of PCT and hsCRP in the death group were higher than those in the survival group.The areas under ROC curve of PCT and hsCRP for diagnosis of sepsis and septic shock as the optimal cut-off point at ≥ 2 ng/mL and at ≥ 75 mg/L,had the sensitivity of 62.1％ and 81.2％,respectively,and the specificity of 89.2％ and 68.2％,respectively.Conclution PCT and hs CRP levels have a certain value in assessing the severity of pneumonia and sepsis patients as well as prognostic evaluation.

Objective To explore the relationship between left atrial diameters (LAD) and prothrombotic state in senile patients with hypertension (HT) and atrial fibrillation (AF). Mcthods Totally 105 patients with cssential hypertension were divided into 65 patients with atrial fibrillation and 40 cases without atrial fibrillation,and then patients with atrial fibrillation were subgrouped into paroxysmal and persistent atrial fibrillation groups.30 healthy people without hypertension and atrial fibrillation were used as control group.LAD was determined by M type ultrasound cardiogram.Fibrinogen (Fg),D-Dimer(D-Dimer),von willebrand (vwF) and haematocrit (HCT) were also measured as prothrombotic state and compared among the groups. Results In groups of HT with AF versus HT without AF versus control,LAD[(43.56 ± 6.72) mm vs.(36.28 ± 5.83) mm vs.(31.63±4.32)mm],Fg[(4.24±0.59)g/L vs.(3.09 ±0.49)g/L vs.(2.80± 0.46)g/L],D-Dimer [(0.43±0.13)mg/L vs.(0.28±0.]0)mg/L vs.(0.18±0.08)mg/L],vwF[(290.44±29.02)％ vs.(101.32±21.36)％ vs.(84.15±20.26) ％],HCT[(0.46±±0.07)vs.(0.37±0.05)vs.(0.34±0.03)]were significantly higher in HT patients with atrial fibrillation than those without atrial fibrillation and control ( all P＜ 0.05),and there were differences in LAD and D-Dimet (P＜＜0.05),but not in Fg,vwF and HCT (all P＞0.05) between patients without atrial fibrillation and control.LAD[(46.75±7.32)mm vs.(40.82±6.21)mm],Fg [(4.68±0.65)g/L vs.(3.85±0.53)g/L],D-Dimer [(0.48±0.16)mg/L vs.(0.40±0.12)mg/L],vwF [(384.96±29.75)％ vs.(209.43±28.63)％] and HCT [(0.49±0.08)vs.(0.43±0.06)] in persistent atrial fibrillation group were increased than those in paroxysmal atrial fibrillation group ( P ＜ 0.05 ).Fg ( r =0.683 ),D-Dimer ( r =0.735 ),vwF ( r=0.763) and HCT(r=0.759)were corrclated with LAD (all P＜0.01). Conclusions Increased LADmight he one of the elevated risks of atrial fibrillation and a higher prothrombotic state is increasing with larger LAD in senile hypertension.

BACKGROUND:Monofocal and multifocal Toric intraocular lens that have been widely used in clinic exhibit xcelent biological and optical characteristics and have good safety and stability after implantation. OBJECTIVE:To compare the outcomes and rotation stability in patients with cataract and astigmatism after implantation of monofocal and multifocal intraocular lens. METHODS:A total of 210 patients with cataract and astigmatism who received phacoemulsification and intraocular lens implantation were included in this study. Of them, 105 patients were assigned to monofocal intraocular lens implantation and the other 105 patients to multifocal intraocular lens implantation. Uncorrected visual acuity, best corrected visual acuity, residual astigmatism were reexamined at 1, 3 weeks and 1 month after surgery. The rotation of Toric intraocular lens was determined. The incidence of complications and spectacles- independent rate were recorded. RESULTS AND CONCLUSION:Visual acuity and residual astigmatism in each group were significantly improved after 1 week of intraocular lens implantation (P < 0.05); furthermore, these two indicators became better over time. Improvement of visual acuity and residual astigmatism in multifocal intraocular lens group was more obvious than that in monofocal intraocular lens group. Postoperative intraocular lens rotation at < 5° occurred in both groups. The intraocular lens rotation degree in multifocal intraocular lens group was higher than that in monofocal intraocular lens group at different time points (P < 0.05). There were no significant differences in incidence of complications and spectacles-independent rates between two groups at 1 month after surgery. These results demonstrate that multifocal Toric intraocular lens provides better visual acuity and residual astigmatism improvement, while monofocal Toric intraocular lens provides better rotation stability.

Objective To evaluate the effects of morphine preconditioning on the expression of microRNAs (miRNAs) during hypoxia-reoxygenation (H/R) in isolated cardiomyocytes in rats with heart failure.Methods Healthy adult male Sprague Dawley rats,w eighing 200-220 g,were used in this study.Adriamycin 2.0 mg/kg was injected once a week for 6 weeks via the tail vein to induce heart failure.The cardiomyocytes were isolated from the failing hearts of rats and seeded in 24-well plates or in 60 mm diameter dishes.The cells were then randomly divided into 3 groups (n =16 each) using a random number table:control group (group C); group H/R;morphine preconditioning group (group MP).The cells were cultured in normal culture atmosphere in group C.After being exposed to hypoxic air (5％ CO2-95％ N2) for 90 min,the cells were returned to the high-glucose DMEM supplemented with 10％ newborn bovine serum and were then cultured for 120 min in H/R and MP groups.In group M,the cells were cultured in morphine culture medium (final concentration of morphine 0.3 μmol/L) for 10 min and then were returned to the culture medium without morphine and cultured for 30 min immediately before hypoxia.At 120 min of reoxygenation,the cells of 8 wells in each group were chosen to detect the cell viability and lactate dehydrogenase (LDH) activity (by Typan blue staining).All the RNAs were extracted from the cardiomyocytes of the left 8 wells in each group and subjected to miRNA microarray to screen differentially expressed miRNAs.Results The cell viability was significantly lower,the activity of LDH was higher,the expression of miR-6216 and let7e-5p was higher,and the expression of miR-133b-5p was lower in H/R and MP groups than in group C (P ＜ 0.05).Compared with H/R group,the cell viability was significantly increased,the activity of LDH was decreased,the expression of miR-133b-5p was up-regulated,and the expression of miR-6216 and let-7e-5p was down-regulated in MP group (P ＜ 0.05).Conclusion Morphine preconditioning reduces H/R injury to isolated cardiomyocytes in rats with heart failure through regulating the expression of miRNAs such as miR133b-5p,miR-6216 and let-7e-5p.

During the process of China’s healthcare reform, the concept of “costly access to health care” has been discussed for quite a long time. However, big data indicates that a small number of patients (5%) account for disproportionate costs ( approximately 50%) . “Costly access to health care” may not be a ubiquitous perceived prob-lem for all residents. In this paper, the concept“high-need patient” will be introduced and the characteristics of the population will be analyzed. According to the healthcare reform experience from the international perspective, this paper proposed healthcare strategy for Chinese high-need patients and policy recommendations for China’s healthcare reform.

Objective: To investigate the effect and mechanism of helix B surface peptide (HBSP) on myocardial ischemia reperfusion injury (MIRI) in experimental mice.
Methods: The MIRI model was established by ligation of anterior descending coronary artery of the mice for 45 min and followed by corresponding treatment at 5 min before reperfusion. A total of 64 male ICR mice were randomly assigned to 4 group:①Sham group,②MIRI group, the mice received normal saline at 5 min before reperfusion,③HBSP group, MIRI mice received HBSP at 5 min before reperfusion and④HBSP+PD98059 group, MIRI mice received PD98059 (a speciifc blocker of ERK1/2) at 20 min before reperfusion and followed by HBSP at 5 min before reperfusion.n=16 in each group, all animals were treated for 2 hours. The area of myocardial infarction (MI) was detected by TTC-EB double staining method, the myocardial apoptosis rate was examined by TUNEL method, the levels of protein expression of ERK1/2 and phosphorylation of ERK1/2 were measured by Western blot analysis.
Results: Compared with MIRI group, HBSP group presented decreased MI area, decreased myocardial apoptosis rate and increased phopsphorylation level of ERK1/2, allP<0.05. Compared with HBSP group, HBSP+PD98059 group showed decreased phopsphorylation level of ERK1/2, increased myocardial apoptosis rate and increased MI area, allP<0.05.
Conclusion: HBSP may reduce the MI area via inhibiting myocardial apoptosis and therefore, protecting the experimental mice from MIRI; the mechanism might be related to the activation of ERK1/2 pathway.

Objective: To explore the points selection pattern of acupuncture for sleep apnea syndromes by data mining technique. Methods: Clinical literature about acupuncture therapy for sleep apnea syndromes was derived from China National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP), PubMed and Science Direct between the time that databases were created and March 25th,2017. Relevant excel database was established and descriptive studies and association rules were analyzed. Results: The most frequently used point was Lianquan (CV 23) and the most frequently used meridian was the Stomach Meridian. The analysis of association rules showed that the clinical choice of acupuncture points was highly correlated, among which the combination of the highest degree of confidence and the highest degree of support was Shenmen (HT 7) and Sishencong (EX-HN 1); Lieque (LU 7), lianquan (CV 23) and Zhaohai (KI 6). Conclusion: Acupuncture treatment of sleep apnea syndromes has specific selection rules of points, providing certain references for clinical and scientific research.

Objective:To explore the safety and efficacy of Castor branched stent-graft exclusion in the treatment of aortic dissection and aneurysm involving left subclavian artery.Methods:The clinical and imaging data of 88 patients with aortic dissection or aneurysm involving left subclavian artery diagnosed by CTA or DSA in our hospital from December 2017 to December 2019 were collected retrospectively, including 67 aorta dissections, 7 thoracic aortic aneurysms and 14 aorta penetrating ulcer. All patients were treated with branched stent-graft under the guidance of DSA. The postoperative curative effect and complications were observed. The paired t test was used to compare the maximum aortic diameter of the lesion before and 6 months after the operation. Results:Eighty-eight patients were successfully treated with integrated stent, the success rate of operation was 100%, and the disease-related symptoms of all patients were basically or completely relieved. The mortality rate within 1 month after the operation was 2.7% (2/88). The two deaths were complicated with other serious diseases before the operation, and the cause of death was not related the operation. All patients were followed up except 4 patients who lost contact after discharge. During the follow-up, there were 1 case of retrograde type A dissection, 1 case of new aneurysm of aortic arch, 2 cases of in-stent stenosis of left subclavian artery branch, 3 cases of mild stroke, no persistent endoleak and no death or other serious complications. The mean maximum aortic diameter at 6 months after operation [(34±4)mm] was significantly lower than that before operation [(38±6 mm)] ( t=6.63, P<0.05). Conclusion:Castor branched stent-graft is simple, mini-invasive and effective in the treatment of aortic dissection and aneurysms involving the left subclavian artery.