Objective To construct yeast eukaryotic expression vector carrying human endostatin (ES) cDNA. Methods The functional fragment of endostatin gene in human hepatic tissue was amplified by using RT-PCR technology, and cloned into yeast pPIC9 expression vector. The positive clone was sequenced by using automatized sequencer. Results The endostatin cDNA was successfully cloned. The positive ES clone gene in pPIC9 expression vector was sieved, and its coding sequence was identified to be as same as the previously reported sequence. Conclusion The successful construction of ES gene in pPIC9 expression vector using molecular biological method maybe helpful for the high expression of ES protein, which may lay the foundation for the treatment of malignant tumor through anti-angiogenesis appoach.

To obtain an ideal recombinant C-terminal fragment of the merozoite surface protein of Plasmodium falciparum in the Pichia pastoris expression system, the major surface protein-119 (MSP-119) gene sequence bearing the 6-his gene was inserted into expression vector pPIC9k and the target gene was transformed to the susceptible yeast cells GS115 by using electroporation. The multiple inserts were screened and the successfully expressed MSP-119 protein with the relative molecular weight of 12kDa in the supernatants of cell cultures could be detected by SDS-PAGE. Meanwhile, Western blot analysis also demonstrated that this protein reacted with mouse anti-MSP-119 monoclonal antibody, and the expression level of MSP-119 was more than 1.0 g/L. It is concluded that this recombinant protein expressed in the Pichia pastoris expression system resembles the native proteins existed.

By means of literature review,expert interviews and experts sorting,the authors identified three approaches of medical humanities care,and applied them to three clinical departments of top ten incidence of medical disputes of the hospital in 2012.These departments have similar number of beds,and covered both internal medicine and surgery departments.The purposes of the study are to evaluate the effects of medical humanities care measures in clinical departments,and to make relevant thoughts and suggestions.

Using the method of semi -structure interviews to pre-interview 6 experts and officially interview10 experts , then the record of the interview was compiled into the paper .Using the grounded theory method to encode material of the interview , a summary was made for each case base on data collation and analysis .This paper ana-lyzed the humanities concern the connotation of the application in the construction of a harmonious doctor -patient relationship and the specific measures .On the basis of the application of humanistic care in the construction of a harmonious doctor -patient relationship , significance and limitation is discussed .

Objective:To study the metabolic mechanism of protective effect of waternut herb extract on primary Aβ SAMP8 damage of mouse hippocampal neurons by metabolic footprinting.Methods:MTT assay was used to determine the proliferation of primary hippocampal neurons in SAMP8 mice with Aβ damage,the effect for the first time on the basis of metabolic footprinting evaluation waternut herb extract.Focus on key metabolic pathways and related metabolic targets,mechanism of primary Aβ SAMP8 damage of mouse hippocampal neurons and pathogenesis of watemut herb extract.Results:MTT assay was used to measure the rate of cell proliferation.The results showed that the cell viability of the primary hippocampal neurons was significantly decreased in the Aβ SAMP8 mice.The study found that metabolic footprinting,compared with littermate wild-type mice,neuronal cell metabolism Aβ SAMP8 damage of mouse anomalies mainly concentrated in the metabolism of folic acid and taufine metabolism associated with nerve cells,by high-throughput mass spectrometric analysis and literature database retrieval to determine the 3 differential metabolites,respectively is L-disodoum alanine (L-Cysteic acid),dihydrofolate (Dihydrofolate),acid (Chorismate),the branch of small molecule metabolites through extract intervention after Amakusa callback trend obviously.Conclusion:the therapeutic effect of watemut herb extract on Aβ SAMP8 damage of mouse primary hippocampal neurons to a certain extent,3 biomarkers of this discovery may be a potential target of Aβ SAMP8 damage of mouse primary hippocampal neurons in the pathogenesis of waternut herb extract,given after these markers were callback trend in different degree,suggesting that watemut herb extract could regulate metabolism related enzymes and metabolic pathways to protect the purpose,to provide the experimental basis for the treatment of Alzheimer's disease watemut herb extract.

Objective To explore the imaging findings and diagnostic values of X-ray, CT, MR,and ultrasonography in traumatic knee joints hemarthrosis and lipohemarthrosis. Methods Traumatic knee joints hemarthrosis (12 knees) and lipohemarthrosis (18 knees) proved by operation (27 knees) or puncturation (3 knees) were included in the study. Horizontal-beam plain radiographs (16 knees), CT (30 knees), MRI (30 knees) and ultrasonography (24 knees) in supine position were investigated. Results (1)supine position horizontal-beam plain radiographs: Fat-liquid layer was found in 8 cases of lipohemarthrosis. Dense supragenual bursa was found in 1 case of lipohemarthrosis and 7 cases of hemarthrosis. Fracture (13 knees) was diagnosed correctly. (2) CT findings: double fluid-fluid layer was found in 11 of all 18 cases, and single fluid-fluid layer was found in 7 of 11 cases of lipohemarthrosis. Single fluid-fluid layer was found in 3 of 12 cases of hemarthrosis. Isodensity was detected in 9 cases, and high-density blood clot was found in 4 cases. Fracture (30 knees) was diagnosed correctly. (3) MRI findings: in 12 of 18 cases of lipohemarthrosis, double fluid-fluid layer was shown including supernatant layer as short T1, long T2signal and low signal after fat-suppression, middle layer as long T1, long T2 signal and high signal after fat-suppression, and dependent layer as iso-T1, iso-T2 and slight high signal after fat-suppression. Single fluid-fluid layer was seen in 6 cases, only had aforementioned upper and under layer.Only aforementioned supernatant layer and dependent layer were seen in 12 cases of hemarthrosis. 4 cases showed entire blood clot in fluid, T1WI showed middle signal or center iso-signal accompanied with peripheral high signal ring, and fat-suppression imaging showed high signal. T2WI and fat-suppressionimaging showed middling or high signal accompanied with peripheral low signal ring. Fracture (30 knees) was diagnosed correctly. (4) Ultrasound findings: In 10 of 14 cases of lipohemarthrosis, double fluid-fluid level was shown, supernatant layer as equal echo, middle layer as echoless, and dependent layer as cloudy echo. Four cases with single fluid-fluid level only showed aforementioned upper and under layer. Three of 10 cases of hemarthrosis showed single fluid-fluid level, only showing aforementioned upper and under layer,and 7 cases showed cloudy echo and float. In 3 cases the fluid blood clot showed irregular shape low-equal echo bolus. No fracture hne was found. Conclusions CT can clearly detect fracture line, hemarthrosis and lipohemarthresis, and can substitute plain radiography. MRI is the best way to diagnose hemarthresis and lipohemarthrosis. Ultrasonography can be used in diagnosing hemarthresis and lipohemarthrosis but not helpful in the diagnosis of fracture.

Objective: To investigate the incidence of small intestinal bacterial overgrowth (SIBO) and systemic low-grade inflammation in patients with irritable bowel syndrome (IBS). Methods: From June to October in 2017, 50 cases of IBS patients who met Rome Ⅳ criteria were consecutively collected at Outpatient Department of Gastroenterology of Shanghai Huashan Hospital. The incidence of SIBO was detected by hydrogen lactulose breath test (LBT) and methane LBT. The incidence of systemic low-grade inflammation in IBS patients was determined by fractional exhaled nitric oxide(FeNO) breath test. Chi-square test was used for statistical analysis. Results: Among 50 IBS patients, the positive rate of FeNO was 70%(35/50), and the number of FeNO positive cases in diarrhea-predominant (n=28), constipation-predominant (n=14) and mix-type (n=8) IBS paitents was 18, 11 and six, respectively, and the difference was not statistically significant among three groups (χ²=1.020, P=0.600). The incidence rate of SIBO was 60% (30/50), with 20 cases (40%) being only positive for hydrogen LBT, seven cases (14%) being methane LBT, and three cases (6%) being both positive. The numbers of hydrogen LBT and methane LBT in diarrhea-predominant, constipation-predominant, and mix-type IBS patents were 17, three, three and two, six, two, respectively. There were statistically significant differences in positive rates of hydrogen LBT and methane LBT among three groups (χ²=6.076 and 6.392, both P<0.05). The positive rate of FeNO in IBS patients with SIBO was higher than that of IBS patients without SIBO (90%, 27/30 vs. 40%, 8/20), and the difference was statistically significant (χ²=14.286, P<0.01). Conclusions Combination of hydrogen LBT and methane LBT has a higher detection rate of SIBO than traditional single hydrogen LBT. There is a correlation between SIBO and systemic low-grade inflammation in IBS patients.

Objective: To investigate the efficacy of low dose and short-term oral rifaximin in patients with small intestinal bacterial overgrowth (SIBO) related irritable bowel syndrome (IBS). Methods: From June 2017 to June 2018, at the Department of Gastroenterology of Huashan Hospital, Fudan University in Shanghai, a total of 37 patients with SIBO related IBS were sequentially enrolled and divided into three groups: diarrhea type, constipation type and mixed type. All the patients received rifaximin 200 mg each time, three times per day for 14 days. The clinical efficacy before and after treatment were compared by the scores of irritable bowel syndrome symptom severity scale (IBS-SSS) and irritable bowel syndrome associated quality of life (IBS-QoL). The efficacy of rifaximin on SIBO clearance and SIBO related chronic low-grade inflammation was evaluated by lactulose breath test (LBT) and exhaled nitric oxide (eNO). T test and variance analysis were used for statistical analysis. Results: Among 39 patients with SIBO related IBS, 24 patients were diarrhea type, seven were constipation type and six were mixed type. Except one patient quitted the study because of chest tightness and palpitation, the IBS-SSS score of the left 36 patients before treatment was (250.83±55.10), and decreased to (151.11±33.96), and the difference was statistically significant (t=13.686, P<0.01). Before treatment the score of IBS-QoL was (28.03±16.16), and decreased to (14.39±9.31) after treatment, and the difference was statistically significant (t=6.867, P<0.01). There was no significant difference in IBS-SSS and IBS-QoL scores among the diarrhea type, constipation type and mixed type groups (all P>0.05). After treated by rifaximin, the negative conversion rate of SIBO was 52.8%(19/36). The negative conversion rate of hydrogen LBT was 54.5%(12/22) and among 11 methane LBT positive patients, six cases turned negative; and one of three patients with both positive hydrogen LBT and methane LBT turned negative. The negative conversion rate of eNO was 41.7%(15/36). Conclusions Low dose and short term rifaximin treatment can improve the severity of clinical symptoms and quality of life in SIBO-related IBS patients, and the efficacy is not related with the subtypes of IBS.

With the rapid development and widespread application of artificial intelligence （AI） technology， it has brought convenience to people’s lives and work， while bringing a series of ethical challenges such as lack of transparency， privacy leakage， limited autonomy in decision-making， and blurred responsibility attribution. By comparing the ethical risk governance of the application of AI in the medical field， this paper systematically analyzed 30 policy regulations issued by 10 countries and international organizations， revealing the consensus and differences in ethical governance of medical AI internationally. The results showed that transparency， fairness and justice， non-harm， privacy protection， freedom and autonomy， responsibility， and people-oriented principles were seven internationally recognized ethical principles， providing important guidance for building a safe， trustworthy， and humane-respecting medical AI ecosystem. In addition， this paper deeply compared the risk assessment methods in the guidelines of China， the United States， and the European Union， as well as found differences in risk types， risk grading， and risk assessment process standards. These differences not only reflect diversities in different cultural， legal backgrounds， and regulatory concepts， but also highlight the necessity of strengthening international cooperation in the context of globalization. Therefore， it is recommended to develop ethical risk governance standards and norms for medical AI that applicable to China’s national conditions. Based on drawing on international experience， proactively carry out risk assessments， reduce ethical challenges， and promote the healthy and rational application of medical AI technology.

Objective To investigate the expression of serum long non-coding RNA MEG3 and microRNA(miR)-195-5p in patients with severe necrotizing pancreatitis and their relationship with the severity and prognosis of severe necrotizing pancreatitis.Methods A total of 122 patients with acute pancreatitis admitted to Cangzhou Central Hospital from October 2020 to January 2023 were selected as the research objects.Ac-cording to the severity of the disease,the patients were divided into severe necrotizing pancreatitis(severe group,53 cases)and non-severe necrotizing pancreatitis(non-severe group,69 cases).According to the prog-nosis of alternate ending with severe necrotizing pancreatitis can be divided into good prognosis group(38 ca-ses)and poor prognosis group(15 cases).At the same time,50 healthy people who underwent physical exami-nation in the hospital during the same period were selected as the control group.The clinical data and serum levels of lncRNA MEG3 and miR-195-5p in each group were compared.Spearman correlation analysis was used to analyze the relationship between serum levels of lncRNA MEG3,miR-195-5p and acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)and Ranson scores.Multivariate Logistic regression was used to analyze the influencing factors of poor prognosis in patients with severe necrotizing pancreatitis.The receiver operating characteristic(ROC)curve was used to analyze the value of lncRNA MEG3 and miR-195-5p in eval-uating the prognosis of patients with severe necrotizing pancreatitis.Results There was no significant differ-ence in age,gender,body mass index,underlying disease and etiology between severe group and non-severe group(P>0.05).Compared with the non-severe group,APACHEⅡ and Ranson scores were significantly in-creased in the severe group(P<0.05).Compared with the control group,the serum levels of lncRNA MEG3 and miR-195-5p in the non-severe group and the severe group were decreased(P<0.05),and the serum levels of lncRNA MEG3 and miR-195-5p in the severe group were lower than those in the non-severe group(P<0.05).Spearman correlation analysis showed that serum levels of lncRNA MEG3 and miR-195-5p in AP pa-tients were negatively correlated with APACHEⅡ and Ranson scores(P<0.05).Multivariate Logistic re-gression analysis showed that APACHEⅡ and Ranson scores and serum levels of lncRNA MEG3 and miR-195-5p were independent risk factors for poor prognosis in patients with severe necrotizing pancreatitis(P<0.05).ROC curve results showed that the area under the curve(AUC)of lncRNA MEG3 and miR-195-5p for evaluating the poor prognosis of patients with severe necrotizing pancreatitis was 0.767 and 0.777,respectively,the sensitivity was 86.7%and 80.0%,and the specificity was 49.9%and 45.8%,respectively.The AUC of combined e-valuation was 0.982,and the sensitivity and specificity were 86.7%and 78.8%,respectively.Conclusion The serum levels of lncRNA MEG3 and miR-195-5p are related to the severity and prognosis of severe necrotizing pancreatitis,which can evaluate the severity and predict the prognosis of severe necrotizing pancreatitis.