The comprehensive capacity of medical students means mainly the medical operating skill, and the ability toresolve the practical troubles by integrating the knowledge of multisubject, and creative capability. For a long time, inthe teaching of many madical colleges, importance has been attached to imparting the basic theory and knowledge,rather than to fostering the comprehensive capacity of medical students. In 1999, the Medical College of ShandongUnivesity incorporated the physiological, phamacological, pathophisiological and psycological experiment and inte-grated them as a independent experimental curriculum-the functional science experiment. Many years practice showsthat the functional science experiment is a good platform or carrier to foster the comprehensive capability of madicalstudents.

Objective: To propose updates and amendments for the nitroglycerin tablets quality in the Chinese pharmacopoeia 2020 due to the failure of effective separation of 4 known impurities and nonseparation of free nitrate ion and excipients.
Methods: Related substances were analyzed using gradient elution by HPLC, and free nitrate ion was determined on SAX column by different HPLC method.
Results: Using the improved method to test the related substances and free nitrate ion of nitroglycerin tablets,the content of the maximum individual impurity were not more than 0.5%, the total content was not more than 2.4% and the content of free nitrate ion was not more than 6.3%.
Conclusion: The improved method is accurate and feasible. It provided a reference for the updates and amendments of the quality standard of nitroglycerin tablets in the Chinese Pharmacopoeia 2020.

Objective:To investigate the serum hepcidin level and risk factors associated with peripheral arterial disease (PAD) and foot ulcer in type 2 diabetic patients.Methods:From January 2019 to June 2019, 70 patients with type 2 diabetes in Department of Endocrinology of Xiangya Third Hospital were selected, including 21 newly diagnosed patients with type 2 diabetes (DM group), 23 patients with lower extremity vascular disease (PAD group) and 26 patients with foot ulcer (DF group). Serum hepcidin was determined by enzyme linked immunosorbent assay (ELISA). The serum levels of hepcidin in different groups were compared, and the correlation between diabetic lower extremity vascular disease and foot ulcer was analyzed.Results:⑴ The hemoglobin, albumin, triglycerides and total cholesterol were significantly lower in DF group compared with PAD and DM groups ( P<0.05), while the DF group patients were with higher white blood cell (WBC) count and high sensitivity C reactive protein (hs-CRP) than patients in PAD and DM groups ( P<0.05). DF group also showed significantly higher WBC, hs-CRP and neutrophil ratio level (NEUT%) than DM group ( P<0.05). The inflammatory indicators of WBC, hs-CRP and NEUT% showed no significant difference between DM group and PAD group ( P>0.05). ⑵ The levels of hepcidin in DF and PAD groups were higher than those in DM group, while that in DF group were higher than those in PAD group ( P<0.05); Hepcidin was positively correlated with systolic blood pressure, WBC count, NEUT% and ferritin ( P<0.05), and negatively correlated with hemoglobin, glycosylated hemoglobin, albumin and 25-hydroxyvitamin D ( P<0.05). ⑶ Binary multivariate logistic regression analysis showed that elevated hepcidin level was an independent risk factor for diabetic foot ulcer [ OR=1.755, 95% CI: 1.063-2.897, P=0.028]. Conclusions:The fluctuation of serum hepcidin level in diabetic patients is related to the stimulation of inflammation, the degree of anemia and the nutritional status, which means it might be an early indicator of inflammation in diabetic patients with peripheral arterial disease. Moreover, the increase of hepcidin is an independent risk factor for diabetic foot ulcers in our study.

Objective To evaluate the influence of the single and multiple intervertebral space radiofrequency ablation nucleoplasty on the sheep cervical spine stability.Methods Twenty healthy adult sheep were randomly divided into single intervertebral space operation group (A group )and multiple intervertebral space operation group ( B group ),each group was subdivided into postoperative 24 hours group ( A1,B1 group ) and post- operative 3 months group ( A2,B2 group),each group had five sheep.Radiofrequency ablation nucleoplasty on sheep C3/4 in the single intervertebral space operation group or C3/4,C4/5 in multiple intervertebral space operation group.Preoperative cervical vertebrae roentgenograms from all samples in neutral,lateral,hyperextension and hyperflexion positions were collected,and 5F pipe was used as the survey mark of ieonography.For the postoperative 24 hours group( A1,B1 group ),the roentgenograms were collected postoperatively 24 hours later,while for post- operative 3 months group (A2,B2 group),they were collected postoperatively 3 months later.The height of operative intervertebral space,horizontal and angular displacement of neighboring vertebral body were measured respectively.Results The roentgenograms showed no any obvious decrease in the height of intervertebral disc and no any increase in horizontal and angular displacement of neighboring vertebral body were observed in single intervertebral space operation group (A group) and multiple intervertebral space operation group (B group ).There was no significant difference between pre - operation and post - operation in them ( P ＞ 0.05 ).Conclusions The stability of the sheep cervical spine had not been affected by the single or multiple intervertebral space radiofrequency ablation nucleoplasty on the basis of X-ray results.The radiofrequency ablation nucleoplasty had no influences on the stability of sheep cervical vertebrae.

Objective To evaluate the feasibility and clinical effectiveness of arthroscopic femoral osteoplasty for cam-type femoroacetabular impingenent(FAI).Methods From October 2007 to April 2009,31 patients with cam-type FAI underwent hip arthroscopy,including 12 males and 19 females with an average age of 34.1 years(range,18-45).Physical examination,X-ray,3D-CT scans were carried out preoperatively to make the definite diagnosis.All patients were treated with hip arthroscopic debridement and femoral osteoplasty.The patients were followed-up at 1 month,3 months,6 months,1 year,and every year post operation.Outcomes were measured with pain visual analogue scale(VAS),ROM of hip,impingement tests,α angle and modified Harris hip score at final follow-up.Results All patients were followed up for average 22.7 months(range,18-36).At final follow-up,impingement tests among all patients were negative,the range of flexion increased from 101.0°±14.2° to 121.0°±11.7°,the range of internal rotation in a position of 90° flexion increased from 5.0°±9.8° to 30.1°±12.7°,α angle decreased from 74.2°±10.7° preoperatively to 44.7°±8.3°,VAS pain score decreased from 6.1±1.1 preoperatively to 0.9±0.7,modified Harris hip score improved from 41.2±5.7 preoperatively to 73.6±4.1,all differences were statistically significant.Conclusion Arthroscopic femoral osteoplasty for cam-type FAI can win the satisfactory achievements for complete excision of bony prominence at anterior-lateral femoral head-neck junction,restoration of the femoral offset,significant relief of clinical symptoms and improvement of hip range of motion.

Aim To observe the damages induced by hydrogen peroxide in cultured bovine cerebral microvascular endothelial cells (BCMEC) and evaluate the protective effects of hydroxyethylpuerarin on hydrogen peroxide-injured BCMEC. Methods BCMEC were cultured and transferred into modified Eagle medium (MEM). The viability of cells was detected by MTT assay. Cell injury was determined by lactate dehydrogenase (LDH) activity in the extracellular medium. Flow cytometry was employed to observe the occurrence of apoptosis. Morphologic changes of cells were visualized under phase contrast and electron occurrence of apoptosis. Hydroxyethylpuerarin was shown to increase the survival rate and decrease the activity of LDH of BCMEC damaged by hydrogen peroxide. Hydroxyethylpuerarin was also found to protect BCMEC against apoptosis induced by hydrogen peroxide. Conclusion Hydrogen peroxide induces BCMEC injury either by apoptosis or through necrosis. Hydroxyethylpuerarin protects BCMEC against hydrogen peroxide-induced injury in a concentration-dependent manner. Its antioxidant effects might be involved as the mechanism protection.

Objective To investigate the enterovirus ( EV)-carrying status and the circulating se-rotypes in healthy children from inner and border areas of Yunnan Province in 2014 and 2015 and to analyze the genetic characteristics of echovirus 6 (ECHO6), ECHO25 and ECHO11 strains. Methods Stool sam-ples were collected from children less than 15 years old living in 6 to 7 counties of 3 inner prefectures/cities and 8 to 9 counties of border prefectures/cities. Altogether 921 samples were collected including 453 sam-ples in 2014 (213 samples in inner counties and 240 samples in border counties) and 468 samples in 2015 (195 samples in inner counties and 273 samples in border counties). Viruses were isolated from the stool samples and their serotypes were identified by gene sequencing. Results The numbers of EV strains isola-ted from the samples collected in inner counties and border counties in 2014 were 20 ( isolating rate:9.39%, 20/213) and 16 (isolating rate: 6. 67%, 16/240), respectively. The overall isolating rate for 2014 was 7. 95% (36/453). The predominant species was enterovirus B, accounting for 88. 89% of all iso-lated strains (32/36), followed by enterovirus A species (11. 11%, 4/36). No strains of enterovirus spe-cies C (including poliovirus) and D was detected in 2014. In total, 46 EV strains were isolated in 2015 with an overall isolating rate of 9. 83% (46/468), including 13 strains in inner counties (isolating rate:6. 67%, 13/195) and 33 strains in border counties (isolating rate:12. 09%, 33/273). Most of the strains were enterovirus B species, accounting for 78. 26% (36/46), followed by enterovirus C species (19. 57%, 9/46) and enterovirus A species (2. 17%, 1/46). Altogether 82 EV strains were isolated in 2014 and 2015 with an isolating rate of 8. 90% (82/921), of which 33 strains were isolated in inner counties (8. 09%, 33/408) and 49 strains were isolated in border counties (9. 55%, 49/513). Among the 82 EV strains, 9 strains were polioviruses (0. 98%, 9/921) and all of them were Sabin-like polioviruses. The rest of the strains were non-polio enterovirus (7. 93%, 73/921). Conclusion In 2014, the EV isolating rate in inner counties (9. 39%) was higher than that in border counties (6. 67%). However, the EV isolating rate in border counties (12. 09%) was higher than that in inner counties (6. 67%) in 2015. Enterovirus B was the predominant species in both 2014 and 2015. No wild type polioviruses and enterovirus D species were detec-ted. Polio-free status was maintained well in Yunnan Province.

Objective:To express human epididymis protein 4 (HE4) in prokaryotic cells,purify the expressed product and de-termine its activity by immunoassay kit.Methods: The gene encoding HE 4 was cloned using RT-PCR technique from total RNA of ovarian carcinoma cells ES-2,the amplified HE4 gene was cloned into prokaryotic expression vector pGEX-4T-1 and PET26b respective-ly.The recombinant plasmid pGEX-4T-1-HE4 and PET26b-HE4 were constructed and transformed into E.coli BL21 cells respectively, and protein ( GST-HE4 and His-HE4 ) expressions were induced by IPTG and identified by SDS-PAGE and commercial ELISA kit.Results:Restriction analysis and sequencing proved that recombinant plasmid pGEX -4T-1-HE4 and PET26b-HE4 were constructed correctly.The expressed recombinant proteins ,with the relative molecular mass of about 38 000 and 12 000 ,showed specific binding to monoclonal antibody against HE 4.Conclusion:Two kinds of recombinant HE 4 protein are successfully expressed in prokaryotic cells , which laid a foundation of preparation of immunoassay reagents.

Objective To evaluate the significance of electronic gastroscopy examination combined with miR-21 in the diagnosis of gastric carcinoma.Methods 96 gastroscopy cases in Guangzhou General Hospital of Guangzhou Military Command were adopted from June,2011 to December,2012.The patients received gastric endoscopy examination,and the diseased tissue samples were taken out at the same time.To calculate the diagnosis rate and missed rate with gastroscopy,by comparing the gastroscopy diagnosis with the pathology diagnosis.To evaluate the expression level of miR-21 of the tissue by applying quantitative real time PCR.Results The diagnosis rates of gastric cancer,gastric ulcer,chronic superficial gastritis,gastroesophageal reflux gastritis,chronic atrophic gastritis under gastroscopy were 28.12 ％ (27/96),30.21％ (29/96),18.75 ％ (18/96),10.42 ％ (10/96),12.50 ％ (12/96).The diagnosis rate of gastric cancer was 100.00 ％ and the missed diagnosis rate was 6.90 ％ (2/29),the relative transcript level of miR-21 in the gastric cancer group was 54.41±6.66,which was obviously higher than the gastric ulcer group and the other groups (P ＜ 0.01).Conclusions Electronic gastroscopy is a clinically practical examination method in screening gastric cancer cases,evaluating the expression level of miR-21 in the diseased tissue under gastroscopy can enhance the accuracy of screening gastric cancer.Combining the two methods is useful in the diagnosis of gastric cancer.

Objective To investigate cartilage oligomeric matrix protein( COMP) gene mutation in a three-generation pedigree with two cases of pseudoachondroplasia, and to definitize genotype-phenotype correlation. Methods The clinical data and peripheral blood were collected from the patients with pseudoachondroplasia and their family members. All the 19 exons and their flanking sequences of COMP gene in two patients and three unaffected family numbers and 50 unrelated individuals were analyzed by PCR amplification and direct sequencing. Results The proband, a 6-year-old girl presented with typical clinical features of pseudoachondroplasia, including disproportionate short limb dwarfism, staggering gait, double genu varus deformity, and wider clinical and imaging long bone metaphysis. The 33-year-old father showed a similar manifestation including disproportionate short limb dwarfism and double genu varus deformity, and was performed correcting operation on lower limbs for double genu varus at the age of 10 years. DNA sequencing analysis of the COMP gene revealed a del mutation ( c. 1417 1419delGAC)in exon 13 in two patients with pseudoachondroplasia, but not in the other unaffected members from the pedrgree and 50 control subjects. Conclusion A del mutation c. 1417 1419delGAC of COMP gene may contribute to the disease in the pedigree.