Objective To explore the role of ripply1 in zebrafish dorsal-ventral development .Methods Using ze-brafish whole-mount in situ hybridization to examine the ripply1 expression pattern in early embryo development .To analyse the expression pattern changes of dorsal-ventral marker genes at shield stage and the morphological changes at 24 hpf (hours post-fertilization) after overexpression of ripply1 by injecting synthetic mRNA at 1-cell stage.Using Tol2 transposon technology to obtain a ripply1 promoter driven GFP transgenic fish and to identify promoter region that recapitulates endoge -nous ripply1 expression pattern .Results The in situ hybridization results revealed that ripply1 specifically expresses in the future dorsal region at shield stage .Overexpression of ripply1 caused an enhanced expression of dorsal marker genes and a reduction of ventral marker genes .Embryos overexpressing ripply1 also showed severely dorsalized phenotype , with enlarged head, reduced ventral yolk extension , and shortened posterior trunk and tail regions , and the formation of a secondary trunk axis.Transgenic fish revealed the maternal expression of ripply1 and suggested that a 1.2 kb promoter-driven GFP is able to recapitulate the endogenous gene expression pattern .Conclusion ripply1 may participate in the early development of dor-sal-ventral axis in zebrafish embryo .

Objective To develop a method for the determination of DBP and DEHP in cosmetics by GC-MS. Methods The samples was extracted by methanol or acetic ether, DBP and DEHP were separated by GC and determined by MS. Results The linear range, regression equation of calibration curve and correlation coefficient were 0.5-100.0 ?g/ml, y=2?106 x-2?106, 0.999 6 for DBP respectively and 5.5-110.0 ?g/ml, y=988 223 x-7?106, 0.995 9 for DEHP respectively. Based on threefold ratio of signal and noise, as the sample was 1.0 and 0.2 g respectively, the volume was 10.0 ml, the detection limits were 1.0 and 5.0 mg/kg respectively for both DBP and DEHP. The recovery rates were 90.8%-119.0% for DBP and 90.4%-115.3% for DEHP. Relative standard deviations were 4.8%-9.8% for DBP and 6.0%-8.6% for DEHP. Conclusion This method is sensitive, accurate and high reproducible, and was applicable to the determination of DBP and DEHP in the cosmetics.

Objective To construct a human endostatin adenovirus vector and investigate its inhibitory effect on pancreatic carcinoma in nude mice.Methods Animal model of pancreatic carcinoma bearing nude mice was established by subcutaneous injection of SW1990 cells.All mice were randomized into Ad-hEnd group,Ad-LacZ group and control group with 8 mice in each group.The endostatin gene recombinant adenovirus were intratumorally injected every two days for 4 times.The rate of tumor growth was observed.lmmunohistochemical staining was employed to investigate the expression of vascular endothelial growth factor (VEGF) and micro-vessel density (MVD).TUNEL in situ was used to examine tumor cell apoptosis.Results The tumor formation rate was 100%.4 weeks later,the volumes of the tumors were (921.9±279.7 )mm3,(2804.4±553.5 )mm3 and ( 3040.6±487.6 ) mm3 in Ad-hEnd group,Ad-LacZ group and control group,respectively;the weights of the tumors were (1.19±0.18 ) g,( 2.38±0.42 ) g and ( 2.41±0.47 ) g,respectively;the VEGF positive rates were (36.3±7.1 )%,(81.2±6.6)% and (79.4±6.2)%,respectively;the levels of MVD were 12±4,27±5 and 25±6,respectively;the apoptotic rates were (31.2 ±5.4) %,( 9.4±4.9 ) % and ( 8.5±3.7 ) %,respectively.Compared with Ad-LacZ group and control group,the parameters in Ad-hEnd group were statistically different (P ＜0.01 ).The difference betweon Ad-LacZ group and control group was not statistically different.Conclusions Human endostatin gene mediated by recombinant adenovirus could inhibit tumor growth,angiogenesis and promote cell apoptosis of pancreatic carcinoma and could be used as geue therapy for pancreatic carcinoma.

Objective To investigate the role of interleukin-17 (IL-17) in spinal dorsal horns in neuropathic pain (NP) in rats and its effect on activation of astrocytes.Methods In vivo experiment Sixty-four male SPF Sprague-Dawley rats,aged 6-8 weeks,weighing 180-200 g,were randomly divided into 3 groups using a random number table:control group (group C,n =16),sham operation group (group S,n =24) and group NP (n =24).The animals were anesthetized with intraperitoneal pentobarbital sodium,the L5,6 spinal nerves of the left side of the rat were gently separated and exposed,tightly ligated with 5-0 silk suture and transected.In group S,the L5,6 spinal nerves of the left side of the rat were only exposed.In group C,no operation was performed.Mechanical pain threshold was measured at day 1 before operation and days 1,3,5,7,10 and 14 after operation.The expression of IL-17,IL-6,IL-1β and tumor necrosis factor-alpha (TNF-α) mRNA in the spinal dorsal horn was determined using quantitative real-time PCR at day 7 and day 14 after operation.At day 7 after operation,the activation of astrocytes in the spinal dorsal horn was detected.In vitro experiment Primarily cultured astrocytes of neonatal rats were randomly divided into 4 groups using a random number table:control group (group C,n=22),10 ng/ml IL-17 group (I10 group,n=18),50 ng/ml IL-17 group (I50 group,n-18) and 100 ng/ml IL-17 group (I100 group,n=22).In I10,I50 and I100 groups,the astrocytes were incubated with the culture medium containing 10,50 and 100 ng/ml IL-17,respectively.The proliferation of astrocytes was detected by MTT at 24,48 and 72 h of incutation or culture.The expression of IL-6,IL-1β and TNF-α mRNA was determined using quantitative real-time PCR.Results In vivo experiment Compared with group C,the mechanical pain threshold was significantly decreased at 3-14 days after operation,the expression of IL-17,IL-6 and IL-1β mRNA in the spinal dorsal horn was up-regualted at 7 days after operation,and the activation of astrocytes was increased in group NP,and no significant change was detected in the mechanical pain threshold at each time point after operation in group S.In vitro experiment Compared with group C,the proliferation of astrocytes was significantly increased at 48 h of incubation in I10 and I50 groups,the proliferation of astrocytes was significantly increased at 48 and 72 h of incubation,and the expression of IL-6 and IL-1β mRNA was up-regulated in I100 group,and no significant change was found in the proliferation of astrocytes in group S.Conclusion Up-regulated expression of IL-17 in spinal dorsal horns may be involved in the maintenance of NP,and the mechanism is related to promoted activation of astrocytes and induced inflammatory responses in rats.

Objective:To explore the independent risk factors of poor healing of surgical incisions in patients with drainage tube removal after thoracic and abdominal surgery and establish a risk prediction model for poor healing of surgical incisions.Methods:Using the convenient sampling method, a total of 545 patients who underwent thoracic and abdominal surgery in the Affiliated Hospital of Qingdao University were selected from July to December 2020. The patients were divided into the poor wound healing group ( n=87) and the non-incision poor healing group ( n=458) according to whether they had poor wound healing. Logistic regression analysis was used to analyze the risk factors of poor healing of surgical incisions and build a risk prediction model. The receiver operating characteristic (ROC) area under the curve was used to test the model to predict the effect and 230 patients were selected to verify the model prediction effect. Results:In this study, 5 factors including duration of exudation, serum albumin, incision infection, the volume of exudation during catheterization and catheterization time were finally included to construct a risk prediction model. The model formula was Z=4.608+4.855× duration of exudation +3.173× serum albumin +3.739× infection of the incision +2.271×the volume of exudation during catheterization + 0.466× catheterization time. The area under ROC curve of this model was 0.773 (95% CI: 0.678 - 0.868). The maximum value of Youden index was 0.549, the sensitivity was 0.742 and the specificity was 0.807. Conclusions:The risk prediction model of poor incision healing after drainage tube removal for patients undergoing thoracic and abdominal surgery can better predict the risk of poor incision healing and provide a basis for clinical medical staff to take preventive management measures for high-risk patients in time.

Objective:To construct a non-invasive pre-hospital screening model and early based on artificial intelligence algorithms to provide the severity of stroke in patients, provide screening, guidance and early warning for stroke patients and their families, and provide data support for clinical decision-making.Methods:A retrospective study was conducted. The clinical information of stroke patients ( n = 53?793) were extracted from the Yidu cloud big data server system of the Second Affiliated Hospital of Dalian Medical University from January 1, 2001 to July 31, 2023. Combined with the results of single factor screening and the opinions of experts with senior professional titles in neurology, the input variable was determined, and the output variable was the National Institutes of Health Stroke Scale (NIHSS) representing the severity of the disease at admission. Python 3.7 was used to build DeepFM algorithm model, and five data mining models including Logistic regression, CART decision tree, C5.0 decision tree, Bayesian network and deep neural network (DNN) were built at the same time. The original data were randomly divided into 80% training set and 20% test set, which were used to train and test the models, adjust the parameters of each model, respectively calculate the accuracy, sensitivity and F-index of the six models, carry out the comprehensive comparison and evaluation of the model. The receiver operator characteristic curve (ROC curve) and calibration curve were drawn, compared the prediction performance of DeepFM model and the other five algorithms. In addition, the data of stroke patients ( n = 1?028) were extracted from Dalian Central Hospital for external verification of the model. Results:A total of 14?015 stroke patients with complete information were selected, including 11?212 in the training set and 2?803 in the testing set. After univariate screening, 14 indicators were included to construct the model, including gender, age, recurrence, physical impairment, facial problems, speech disorders, head reactions, disturbance of consciousness, visual disorders, abnormal cough and swallowing, high risk factor, family history, smoking history and drinking history. DeepFM model adopted the two-order crossover feature. The number of hidden layers in DNN layer was 3. Dropout was used to discard the neurons in the neural network. Rule was used as the activation function. Each layer used Dense full connection. The objective function was random gradient descent. The number of iterations was 15. There were 133?922 training parameters in total. Comparing the predictive value of the six models showed that the accuracy of DeepFM model was 0.951, the sensitivity was 0.992, the specificity was 0.814, the F-index was 0.950, and the area under the curve (AUC) was 0.916. The accuracy of the other five data mining models were between 0.771-0.780, the sensitivity were between 0.978-0.987, the F-index were between 0.690-0.707, and the AUC were between 0.568-0.639. The calibration curve of the DeepFM model was more aligned with the ideal curve than the other five data mining models. Suggesting that the prediction performance of DeepFM model was the best. External validation was conducted on the DeepFM model, and its accuracy was 0.891, indicating good generalization performance of the model.Conclusion:The pre-hospital non-invasive screening prediction model based on DeepFM can accurately predict the severity grading of stroke patients, and has potential application value in rapid screening and early clinical decision-making of stroke.

Objective To analyze the composition of the aqueous extract of Polygonatum Cyrtonema Hua(PCHE)and evaluate its antitumor activity in vitro and in vivo.Our aim is to provide a theoretical basis for the further development and utilization of Polygonatum Cyrtonema Hua.Methods(1)PCHE was prepared by aqueous extraction,and the chemical composition of PCHE was analyzed by UPLC-Q-TOF/MS and phenol-sulfuric acid method.The inhibitory activity on tumor cells proliferation of PCHE was detected by CCK-8 assay.Cell cycle and apoptosis were detected by flow cytometry,and the expression of apoptosis-related proteins Bcl-2 and Bax was detected by Western Blot.The inhibitory activity of PCHE-containing serum on cell proliferation was detected.(2)A B16 tumor-bearing mice model was constructed and model mice were randomly divided into the model group(saline),the positive drug group(CTX:50 mg·kg-1),and PCHE low-,medium-,and high-dose groups(55.9,111.8,223.6 mg·kg-1),and treated by gavage for 7 days.Changes in body weight and tumor volume of mice were observed during the treatment period.The mice were executed after the treatment,and the histopathological changes of heart,liver,spleen,lung,kidney and tumor were observed by hematoxylin-eosin(HE)staining.The protein expression of Bcl-2 and Bax in tumor tissues was detected by immunohistochemistry(IHC).Results The polysaccharide content of PCHE reached(10.07±1.3)%,and the flavonoid content was(0.044±0.004)%,and thirty-nine components were detected by UPLC-Q-TOF/MS,which contained antitumor components such as flavonoids(baicalein,quercetin,luteolin and rutin),organic acids(ferulic acid)and polyphenols(gallic acid),etc.PCHE exhibited the inhibitory effects on Hela,A549,4T1,B16,MFC and HepG2 cells,among which the inhibitory effect on B16 cells was the most significant(P＜0.001),and PCHE induced cell cycle arrest at G0/G1 phase in B16 cells(P＜0.001).The results of double-staining flow cytometry and Western Blot showed that PCHE significantly promoted apoptosis of B16 cells,decreased the expression of Bcl-2,and promoted the expression of Bax(P＜0.01,P＜0.001).and PCHE constituents absorbed into blood also had an inhibitory effect on B16 cells(P＜0.001).In addition,the results of in vivo activity assay showed that different doses of PCHE could inhibit tumor growth,induce tumor cell necrosis,reduce Bcl-2 expression,and increase Bax expression compared with the model group.Conclusion The ingredients in PCHE are abundant.It contains a variety of antitumor active ingredients,which can inhibit tumor growth,induce tumor cell apoptosis,show strong anti-tumor effects and be worthy of in-depth study.

OBJECTIVE：To construc t the evaluation system of the research quality of medical insurance budget impact analysis （BIA），and to provide feasible evaluation tool for related departments as medical insurance department. METHODS ：Based on BIA guidance documents and relevant empirical literatures of ISPOR ，Canada，Poland，the United States and other countries ， combined with expert interview ，the relevant elements of medical insurance negotiation BIA material were confirmed （including key elements and adjuctive elements ）. The scale and system was established to calculate total score of BIA research quality evaluation. RESULTS ：Key elements included three data blocks as target population ，market situation and treatment cost ，involving 14 key elements such as total population ，new drug scenario market share ，treatment cost ，etc.. According to the degree of compliance，0-3 points were assigned and the lowest score after normalization was taken as the basic score of BIA research quality. The adjunctive elements included five data blocks as title & abstract ，research background ，analysis framework ，result presentation and other ，including 23 adjunctive elements such as title ，abstract，research angle ，research time limit ，etc.. According to whether there is quality grade difference ，the elements were divided into type A and type B ；the grade score （0-4 points）and 0/1 score（1 point for yes and 0 point for no ）were used respectively ，and the additional score of BIA research quality was obtained after calculation and addition. According to the addition of different weights （0.67 and 0.33）of basic score and additional score ，the total score system of BIA research quality evaluation could be calculated. CONCLUSIONS ：This study successfully constructed a new BIA quality evaluation system ，which can be used for the quality evaluation of BIA research submitted by medical insurance drug negotiation.

With the increasing proportion of human papilloma virus(HPV)infection in the pathogenic factors of oro-pharyngeal cancer,a series of changes have occurred in the surgical treatment.While the treatment mode has been im-proved,there are still many problems,including the inconsistency between diagnosis and treatment modes,the lack of popularization of reconstruction technology,the imperfect post-treatment rehabilitation system,and the lack of effective preventive measures.Especially in terms of treatment mode for early oropharyngeal cancer,there is no unified conclu-sion whether it is surgery alone or radiotherapy alone,and whether robotic minimally invasive surgery has better func-tional protection than radiotherapy.For advanced oropharyngeal cancer,there is greater controversy over the treatment mode.It is still unclear whether to adopt a non-surgical treatment mode of synchronous chemoradiotherapy or induction chemotherapy combined with synchronous chemoradiotherapy,or a treatment mode of surgery combined with postopera-tive chemoradiotherapy.In order to standardize the surgical treatment of oropharyngeal cancer in China and clarify the indications for surgical treatment of oropharyngeal cancer,this expert consensus,based on the characteristics and treat-ment status of oropharyngeal cancer in China and combined with the international latest theories and practices,forms consensus opinions in multiple aspects of preoperative evaluation,surgical indication determination,primary tumor re-section,neck lymph node dissection,postoperative defect repair,postoperative complication management prognosis and follow-up of oropharyngeal cancer patients.The key points include:① Before the treatment of oropharyngeal cancer,the expression of P16 protein should be detected to clarify HPV status;② Perform enhanced magnetic resonance imaging of the maxillofacial region before surgery to evaluate the invasion of oropharyngeal cancer and guide precise surgical resec-tion of oropharyngeal cancer.Evaluating mouth opening and airway status is crucial for surgical approach decisions and postoperative risk prediction;③ For oropharyngeal cancer patients who have to undergo major surgery and cannot eat for one to two months,it is recommended to undergo percutaneous endoscopic gastrostomy before surgery to effectively improve their nutritional intake during treatment;④ Early-stage oropharyngeal cancer patients may opt for either sur-gery alone or radiation therapy alone.For intermediate and advanced stages,HPV-related oropharyngeal cancer general-ly prioritizes radiation therapy,with concurrent chemotherapy considered based on tumor staging.Surgical treatment is recommended as the first choice for HPV unrelated oropharyngeal squamous cell carcinoma(including primary and re-current)and recurrent HPV related oropharyngeal squamous cell carcinoma after radiotherapy and chemotherapy;⑤ For primary exogenous T1-2 oropharyngeal cancer,direct surgery through the oral approach or da Vinci robotic sur-gery is preferred.For T3-4 patients with advanced oropharyngeal cancer,it is recommended to use temporary mandibu-lectomy approach and lateral pharyngotomy approach for surgery as appropriate;⑥ For cT1-2N0 oropharyngeal cancer patients with tumor invasion depth＞3 mm and cT3-4N0 HPV unrelated oropharyngeal cancer patients,selective neck dissection of levels ⅠB to Ⅳ is recommended.For cN+HPV unrelated oropharyngeal cancer patients,therapeutic neck dissection in regions Ⅰ-Ⅴ is advised;⑦ If PET-CT scan at 12 or more weeks after completion of radiation shows intense FDG uptake in any node,or imaging suggests continuous enlargement of lymph nodes,the patient should undergo neck dissection;⑧ For patients with suspected extracapsular invasion preoperatively,lymph node dissection should include removal of surrounding muscle and adipose connective tissue;⑨ The reconstruction of oropharyngeal cancer defects should follow the principle of reconstruction steps,with priority given to adjacent flaps,followed by distal pedicled flaps,and finally free flaps.The anterolateral thigh flap with abundant tissue can be used as the preferred flap for large-scale postoperative defects.