Large Cell Neuroendocrine Carcinoma Presenting as Adult Intussusception.

Large cell neuroendocrine carcinoma (LCNEC) is an extremely rare malignant tumor of the colon, presenting with more severe clinical outcomes in comparison to colonic adenocarcinoma. There are very few reported cases in the literature. We hereby add our voice to the incidence of this disease by presenting the first report of a patient with ileocolic intussusception secondary to a large cell neuroendocrine cancer of the cecum. The patient was a 48-year-old woman who presented with acute onset of generalized abdominal pain and leukocytosis. CT scan revealed an ileocecal intussusception and multiple liver metastases suggestive of a malignant bowel lesion. She underwent emergency surgery, and an extended right hemicolectomy with ileo-transverse anastomosis was performed. Histology of the resected lesion revealed large cell neuroendocrine carcinoma of the cecum with invasion through the muscularis propria into peri colorectal tissues. The tumor retained mismatch repair (MMR) proteins with low potential for microsatellite instability (MSI). With a clinical diagnosis of stage IV LCNEC, the patient began platinum doublet chemotherapy with carboplatin and etoposide; however, her disease progressed, and the patient expired within a few months after her diagnosis. Clinical diagnosis of adult intussusception should prompt clinicians to rule out malignant etiology. This patient had a large cell neuroendocrine carcinoma of the colon, a rare and extremely aggressive malignancy. Patients with LCNEC will benefit from a multidisciplinary approach to treatment.

Explore of the Red Doctor’s Spirit Integrate into the Ideological and Political Construction of Medical Humanities Course / 中国医学伦理学

The red doctor’s spirit was formed during the medical and health practice led by the Communist Party of China during the Soviet Area of the Communist Party of China. The red doctor’s spirit has distinct characteristics, which is consistent with the value orientation of medical humanities. At the same time, there are some problems in the ideological and political construction of red doctor’s spirit and medical humanities course. It is necessary to explore ways to better integrate red doctor’s spirit into the construction of medical humanities course, so as to promote the improvement and development of medical humanities course. Through the combination of red doctor’s spirit and medical humanities course, this paper developed the ideological and political elements in medical humanities course, expanded the breadth and depth of medical humanities education, so that the majority of medical students can be actively guided in medical humanities course, form firm ideals and beliefs, good moral quality, and achieve the good effect of curriculum ideological and political construction. Guide medical students’ thoughts to develop in the right direction and to lay a solid foundation for medical work in the future.

Spontaneous Pneumothorax in a Young Female With Lymphangioleiomyomatosis.

Lymphangioleiomyomatosis (LAM) is a rare disorder of abnormal proliferation of smooth muscle-like cells which results in the formation of thin-walled cysts and progressive lung destruction. It commonly presents with progressive dyspnea that is often associated with a history of pneumothorax or chylothorax particularly among females of reproductive age. In this report, we present a case of hydropneumothorax as the initial presentation of LAM in a 33-year-old woman, a rather rare presentation. We also discuss the pathogenetic mechanisms, the diagnosis, and treatment strategies using mTOR inhibitors like sirolimus.

Exploring the impacts of economic policy uncertainty, natural resources, and energy structure on ecological footprints: evidence from G-10 nations.

The role of energy and natural resources rent has been studied in the literature but very few research works investigated the impacts of economic policy uncertainty and energy structure on ecological footprints (EFs) in the group of ten G-10 countries. Therefore, this research is designed to probe the impacts of economic policy uncertainty (EP), natural resources rent, energy structure, economic growth, and total factor productivity on EF. The annual data for 1995-2018 is analyzed by adopting second-generation methods. The empirical results from the CS-ARDL unveiled that EP is significantly and positively related to EF, indicating that EP is not environmentally friendly in these nations. A 1% increase in EP contributes to EF by 0.025% in the long run. In addition, the results show that natural resource rents and energy structure positively contribute to mitigating EF by 0.012% and 0.095% respectively, while economic growth increases EF by 0.237%. Based on the empirical results, this work suggests addressing the economic policy uncertainty and economic progress to lower ecological deprivation can be a viable solution to attain sustainable development goals. This work recommends adopting feasible economic policies with sound efforts. For this purpose, the economic sector has to be strong enough with sustainable production. They should ensure the supply of continuous renewable energy to the economic sector.

Exploration of the Path of Integrating Wu Liande’s Spirit into Professional Spirit Education for Medical Students / 中国医学伦理学

Wu Liande’s spirit was formed during the development of modern public health in China. It is a unity of the spirit of patriotism, humanitarianism, scientific exploration, and dedication, and a valuable resource and vivid textbook for medical students to carry out professional spirit education. Promoting Wu Liande’s spirit in the new era is not only conducive to inspiring the patriotism of medical students, bravely undertaking the mission of the times, and devoting themselves to the cause of human health, but also conducive to guiding medical students to refine benevolence and skill, and fulfill the sacred oath of medical students. To cultivate the professional spirit of medical students with Wu Liande’s spirit, it is necessary to achieve the "integration of specialized courses and ideological and political education" , promote the collaborative development of ideological and political courses and curriculum ideological and political education, innovate teaching methods and use modern information technology to empower Wu Liande’s spirit to be visualized and expressed, and take discipline practice as the starting point to expand the new path of professional spirit practice education for medical students.

An Exceptional Case of Diplopia and Ptosis: Extramedullary Plasmacytoma of the Clivus With Multiple Myeloma.

Intracranial plasmacytoma is an exceedingly rare presentation of plasma cell neoplasms. Usually presenting late in the course of the disease, progression from the presentation can be abrupt. Hence, a low threshold to biopsy the lesion should be maintained during diagnostic evaluation. Multiple myeloma workup should also be sent and treated concomitantly along with local treatment. Here, we present a case of extramedullary plasmacytoma of the clivus leading to progressive visual deficits with undiagnosed multiple myeloma requiring pulse steroids, intracranial irradiation, and high-dose chemotherapy with improvement in symptoms.

Research on the Integration of Red Doctor’s Spirit into Medical Ethics Education through Information Technology / 中国医学伦理学

Red doctor’s spirit, an ideological system formed in the revolutionary war era, includes four dimensions of ideal and faith, professional technology, professional value and work style. It is a valuable resource for medical students to carry out medical ethics and medical style education. In the new era, the use of information technology to integrate red doctor’s spirit into medical ethics education not only provides an important guarantee for medical students to "classes suspended but learning continues" under the normalization of the epidemic situation, but also effectively responds to the cognitive needs and expectations of medical students in the context of education informationization, and has cultivated a continuous stream of high-quality medical talents for the implementation. The red doctor’s spirit should be integrated into medical ethics education through technology to ensure proper selection and actively implement the principle of intelligent teaching; adhere to the principle of "content is the most important" and enhance the presentation and ideological guidance of red doctor’s spirit; focus on the innovation of methods to realize the activation presentation and visual expression of red doctor’s spirit.

Improvement and application of radiation work permit software in nuclear power plants / 中国辐射卫生

Objective To improve the efficiency of radiation work preparation, promote the comprehensive and whole-process control of radiation risk, strengthen the accumulation of experience feedback data, and improve the timeliness of approval. Methods According to the application of radiation work permits in Tianwan Nuclear Power Plant and other nuclear power plants, the radiation work permit software in Tianwan Nuclear Power Plant was improved in terms of work order connection, examination and approval process integration, approval of radiation work at different levels, unification of level classification standard, radiation risk database establishment and maintenance, and closed loop management and mobile approval of radiation work permit. Results A radiation work permit software for nuclear power plant was developed according to the requirement of radiation risk control. Conclusion The radiation work permit software effectively improves the radiation work regarding work preparation efficiency, risk control, experience feedback data accumulation, and timeliness of approval. Subsequently, information transmission efficiency and experience sharing can be improved through connection and communication with other software in nuclear power plants.

Advanced Squamous Cell Carcinoma of Gall Bladder Masquerading as Liver Abscess With Review of Literature Review on Advanced Biliary Tract Cancer.

Advanced biliary tract carcinoma (ABC) tends to have a poor prognosis, with trials done having limited data from oncologists' perspectives. Squamous cell variant of gallbladder cancer (GBC) is one of the rarest forms of cancer known in the literature, with a very aggressive course and dismal prospects. Herein, we present a case of a 67-year-old man who got diagnosed with squamous cell carcinoma, which initially masqueraded as liver abscess and was associated with severe hypercalcemia, pyrexia, jaundice, and submassive pulmonary embolism.

Investigation of morphological changes of HPS membrane caused by cecropin B through scanning electron microscopy and atomic force microscopy

Background@#Antimicrobial peptides (AMPs) have been identified as promising compounds for consideration as novel antimicrobial agents. @*Objectives@#This study analyzed the efficacy of cecropin B against Haemophilus parasuis isolates through scanning electron microscopy (SEM) and atomic force microscopy (AFM) experiments. @*Results@#Cecropin B exhibited broad inhibition activity against 15 standard Haemophilus parasuis (HPS) strains and 5 of the clinical isolates had minimum inhibition concentrations (MICs) ranging from 2 to 16 μg/mL. Microelectrophoresis and hexadecane adsorption assays indicated that the more hydrophobic and the higher the isoelectric point (IEP) of the strain, the more sensitive it was to cecropin B. Through SEM, multiple blisters of various shapes and dents on the cell surface were observed. Protrusions and leakage were detected by AFM. @*Conclusions@#Based on the results, cecropin B could inhibit HPS via a pore-forming mechanism by interacting with the cytoplasmic membrane of bacteria. Moreover, as cecropin B concentration increased, the bacteria membrane was more seriously damaged. Thus, cecropin B could be developed as an effective anti-HPS agent for use in clinical applications.

Investigation of morphological changes of HPS membrane caused by cecropin B through scanning electron microscopy and atomic force microscopy

Background@#Antimicrobial peptides (AMPs) have been identified as promising compounds for consideration as novel antimicrobial agents. @*Objectives@#This study analyzed the efficacy of cecropin B against Haemophilus parasuis isolates through scanning electron microscopy (SEM) and atomic force microscopy (AFM) experiments. @*Results@#Cecropin B exhibited broad inhibition activity against 15 standard Haemophilus parasuis (HPS) strains and 5 of the clinical isolates had minimum inhibition concentrations (MICs) ranging from 2 to 16 μg/mL. Microelectrophoresis and hexadecane adsorption assays indicated that the more hydrophobic and the higher the isoelectric point (IEP) of the strain, the more sensitive it was to cecropin B. Through SEM, multiple blisters of various shapes and dents on the cell surface were observed. Protrusions and leakage were detected by AFM. @*Conclusions@#Based on the results, cecropin B could inhibit HPS via a pore-forming mechanism by interacting with the cytoplasmic membrane of bacteria. Moreover, as cecropin B concentration increased, the bacteria membrane was more seriously damaged. Thus, cecropin B could be developed as an effective anti-HPS agent for use in clinical applications.

Clinical study on the treatment of anorexia in children by Tui Na therapy combined with self-made Jianpi-Tiaogan Decoction / 国际中医中药杂志

Objective:To evaluate the efficacy of Tui Na therapy combined with self-made Jianpi-Tiaogan Decoction in the treatment of anorexia in children. Methods:A total of 92 patients with pediatric anorexia in our hospital from January 2018 to June 2019 who met the inclusion criteria were divided into 2 groups by random number table method, with 46 patients in each group. The control group was treated with the Tui Na therapy mainly by manipulating on the abdomen and pushing the spleen, and the treatment group was given the self-made Jianpi-Tiaogan Decoction on the basis of the treatment of the control group. Both groups were treated for 4 weeks. The TCM syndrome scores were performed, serum Leptin and Ghrelin levels were measures by enzyme linked immunosorbent assay, serum neuropeptide Y (NPY) was measured by radioimmunoassay, contents of blood zinc and salivary amylase ptyalase were measured by flame atomic absorption spectrometry and Western blot, and D-xylose absorption test was carried out respectively before and after treatment. And the clinical efficacy was evaluated. Results:The total effective rate was 91.3% (42/46) in the treatment group and 73.9% (34/46) in the control group. The difference between the two groups was statistically significant ( χ2=4.821, P=0.028). After treatment, the scores of anorexia, antifeedance, irritability, agitation, hiccup and belching, fullness of chest and flank, thin shape, sleep difficulty and total score in the treatment group were significantly lower than those in the control group ( t values were 6.782, 7.524, 9.100, 5.643, 4.796, 6.066, 4.069, 3.197, 5.073, respectively, all Ps<0.01). After treatment, the serum NPY (133.7 ± 12.4 ng/L vs. 105.6 ± 10.8 ng/L, t=11.589) and Ghrelin (329.6 ± 26.1 ng/L vs. 275.4 ± 25.0 ng/L, t=12.211) levels in the treatment group were significantly higher than those in the control group ( P<0.01), but Leptin (21.7 ± 4.0 g/L vs. 25.2 ± 4.3 g/L, t=4.042) was significantly lower than that of the control group ( P<0.01). After treatment, blood zinc (9.9 ± 2.5 mol/L vs. 8.7 ± 2.3 mol/L, t=2.396), salivary amylase ptyalase content (23.5 ± 4.2 U/L vs. 17.9 ± 3.6 U/L, t=6.866), D-xylose absorption test (0.9 ± 0.4 ng/L vs. 0.7 ± 0.3 ng/L, t=2.713) were significantly higher than those in the control group ( P<0.05 or P<0.01). Conclusions:Tui Na therapy combined with self-made Jianpi-Tiaogan Decoction can improve TCM syndromes of children, regulate the levels of NPY, Leptin and Ghrelin, and improve clinical efficacy.

Discovery of 1,2,3triazolo4,5-pyrimidine derivatives as highly potent, selective, and cellularly active USP28 inhibitors

Ubiquitin specific peptidase 28 (USP28) is closely associated to the occurrence and development of various malignancies, and thus has been validated as a promising therapeutic target for cancer therapy. To date, only few USP28 inhibitors with moderate inhibitory activity have been reported, highly potent and selective USP28 inhibitors with new chemotypes remain to be discovered for pathologically investigating the roles of deubiquitinase. In this current study, we reported the synthesis and biological evaluation of new [1,2,3]triazolo[4,5-]pyrimidine derivatives as potent USP28 inhibitors. Especially, compound potently inhibited USP28 (IC = 1.10 ± 0.02 μmol/L,  = 40 nmol/L), showing selectivity over USP7 and LSD1 (IC > 100 μmol/L). Compound was cellularly engaged to USP28 in gastric cancer cells. Compound reversibly bound to USP28 and directly affected its protein levels, thus inhibiting the proliferation, cell cycle at S phase, and epithelial-mesenchymal transition (EMT) progression in gastric cancer cell lines. Docking studies were performed to rationalize the potency of compound . Collectively, compound could serve as a new tool compound for the development of new USP28 inhibitors for exploring the roles of deubiquitinase in cancers.

Development of the triazole-fused pyrimidine derivatives as highly potent and reversible inhibitors of histone lysine specific demethylase 1 (LSD1/KDM1A)

Histone lysine specific demethylase 1 (LSD1) has been recognized as an important modulator in post-translational process in epigenetics. Dysregulation of LSD1 has been implicated in the development of various cancers. Herein, we report the discovery of the hit compound (IC = 3.93 μmol/L) and further medicinal chemistry efforts, leading to the generation of compound (IC = 49 nmol/L, and = 16 nmol/L), which inhibited LSD1 reversibly and competitively with H3K4me2, and was selective to LSD1 over MAO-A/B. Docking studies were performed to rationalize the potency of compound . Compound also showed strong antiproliferative activity against four leukemia cell lines (OCL-AML3, K562, THP-1 and U937) as well as the lymphoma cell line Raji with the IC values of 1.79, 1.30, 0.45, 1.22 and 1.40 μmol/L, respectively. In THP-1 cell line, significantly inhibited colony formation and caused remarkable morphological changes. Compound induced expression of CD86 and CD11b in THP-1 cells, confirming its cellular activity and ability of inducing differentiation. The findings further indicate that targeting LSD1 is a promising strategy for AML treatment, the triazole-fused pyrimidine derivatives are new scaffolds for the development of LSD1/KDM1A inhibitors.

Distribution and risk factors of multi drug resistant bacteria in nosocomial infection in Department of Neurosurgery / 中国神经精神疾病杂志

Objective To investigate the clinical features and risk factors of multidrug－resistant bacteria (multi drug resistant organisms, MDROs) infection in Department of Neurosurgery, and to provide evidence for the prevention and control of MDRO infection. Methods Data from 437 cases of infection in hospitalized patients on January 2012－2016 year in December Third Affiliated Hospital of Guizhou Medical University were retrospectively analyzed. Patients were divided into MDROs group and non MDROs group based on the results of MDROs detection. Multi factor Logistic regression analysis model was used to analyze risk factors. Results The infection rate of MDROs was 35.51%, and the detection rate of MDROs was 33.23% . ESBLs, CR－AB and MRSA were the most common bacterial species, and the infection of respiratory tract, urinary tract and wound infection were the main infection sites. Multivariate logistic regression analysis showed that hospitalization time ＞20 d, level of consciousness (coma), occupancy of ICU ≥7 d, ventilation (invasive), number of antibiotics used≥3, combined use of antibiotics≥3, mechanical ventilation Time≥7 d were possible risk factors for MDROs infection in neurosurgical patients (P＜0.05). Conclusion The situation of MDROs infection in neurosurgery is severe. To reduce MDROs infection, it is important to shorten unnecessary hospitalization time, promptly assess and transfer out of ICU as soon as possible, improve microbial examination, avoid frequent change of antibiotics or unnecessary use of use of broad－spectrum antibiotics, reduce unnecessary mechanical ventilation time, change to non－invasive ventilation as far as possible when the condition permits, focus on patients with poor consciousness, and prevent aspiration by mistake.

The construction of recombinant Lactobacillus casei expressing BVDV E2 protein and its immune response in mice.

Bovine viral diarrhea virus (BVDV) is the etiological agent of BVD causes substantial economic losses and endemic in world-wide cattle population. Mucosal immunity plays an important role in protection against BVDV infection and Lactobacillus casei is believed as an excellent live vaccine vector for expressing foreign genes. In this study, we have constructed a novel recombinant L. casei/pELX1-E2 strain expressing the most immunogenic E2 antigen of BVDV; using growth phage dependent surface expression system pELX1. The expression of E2 protein was verified by SDS-PAGE, Western blotting, and Immunofluorescence microscopic analysis. The immune responses triggered by the E2 producing recombinant L. casei were evaluated in BALB/c mice revealed that oral and intranasal (IN) administration of the recombinant strain was able to induce a significantly higher level of specific anti-E2 mucosal IgA and serum IgG as well as the greater level of cellular response by IFN-γ and IL-12 than those of intramuscular (IM) and control groups of mice. However, IN inoculation was found the most potent route of immunization. The ability of the recombinant strain to induce serum neutralizing antibody against BVDV and reduced viral load after viral challenge indicated better protection of BVDV infection. Therefore, this recombinant L. casei expressing E2 could be a safe and promising mucosal vaccine candidate against BVD.

Observation of clinical effect and adverse reaction of hydromorphone for continuous epidural analgesia after cesarean section / 中国基层医药

Objective To learn the clinical effect and adverse reaction of hydromorphone for continuous epidural analgesia after cesarean section,and to provide effective anesthesia method for clinical.Methods 116 cases of cesarean section were randomly divided into observation group(65 cases) and control group(51 cases),continuous epidural analgesia was used in the two groups after operation.Among them,hydromorphone + ropivacaine was used for the observation group,morphine + ropivacaine was used for the control group.The analgesic effect and sedation degree of the two groups were evaluated.The adverse drug reactions were observed,and satisfaction of the patients after the operation was investigated.Results 6,9,24 hours after operation,the VAS pain scores of the observation group were lower than those of the control group,the differences were statistically significant(t=3.236,4.970,2.547,all P0.05).The difference of postoperative first exhaust time between the two groups was not statistically significant(t=0.730,P>0.05).The satisfaction degree of the observation group was significantly higher than that of the control group,the difference was statistically significant(t=2.301,P0.05).Conclusion For continuous epidural analgesia after cesarean section,hydromorphone can effectively relieve the pain after operation,the safety of the process is high,and it has good clinical application value.

Application of ultrasound in evaluating degree of xerostomia in patients with nasopharyngeal carcinoma receiving radiotherapy / 中华放射肿瘤学杂志

Objective To investigate the effectiveness of ultrasound in evaluating the degree of xerostomia in the patients with nasopharyngeal carcinoma ( NPC) receiving intensity?modulated radiotherapy (IMRT). Methods A total of 30 NPC patients who were admitted to our hospital from May 2013 to December 2014 were enrolled in this study. The degree of xerostomia in these NPC patients was scored according to the Radiation Therapy Oncology Group scoring criteria. Color Doppler ultrasound was used to measure the peak systolic blood flow velocity of the parotid gland and submandibular gland and the changes in vascular diameter of the parotid gland in the NPC patients before, during, and after IMRT. The correlation between each parameter and the degree of xerostomia was determined by analysis of variance. Results In the 28 NPC patients included in the statistical analysis, the degree of xerostomia during IMRT was significantly higher than that before IMRT ( P=0024 ) , and the degree of xerostomia at 3 months after IMRT was significantly lower than that during IMRT ( P=0035) . The peak systolic blood flow velocity of the parotid gland and submandibular gland and the proportion of patients with decreases in vascular diameter of the parotid gland during IMRT were significantly higher than those before IMRT ( P=0001 and 0003;P=0001);the above parameters at 3 months after IMRT were significantly lower than those during IMRT ( P=0008 and 0012;P=0001) . During IMRT and after IMRT, the degree of xerostomia was significantly correlated with the peak systolic blood flow velocity of the parotid gland ( r=0563, P=0026;r=0409, P=0031) . Conclusions Ultrasound can be used as a noninvasive detection for the hemodynamic changes in the parotid gland, and it has a certain clinical reference value for evaluating the degree of xerostomia in NPC patients during and after IMRT.

Effects of strut numbers on biomechanical properties of Z-shaped stent-grafts / 医用生物力学

Objective To analyze the influence from strut numbers on biomechanical properties of Z-shaped stentgrafts,especially on their radial support performance and flexibility,so as to provide theoretical support for the design and clinical selection of Z-shaped stent-grafts.Methods Z-shaped stent-grafts with 5,8,10,12 struts were established by finite element method to simulate the process of compression and bending behavior.Radial displacements were applied on surface of the stent-graft and opposite rotations were applied around the z-axis,until a bending angle of 40° was reached.Then parameters such as stent stress,radial support force,strains in graft,cross-section deflection,bending torque were extracted to evaluate the performance of different stentgrafts.Results Compared with stent-grafts with more strut numbers,5-strut stent-graft showed a better radial support performance with the maximum radial force during compression,and it also generated the smallest 68 N · mm torque and 67.5％ cross-section deflection.The tensile strain in graft of 5-strut stent-graft during the process of bending was relatively smaller.Conclusions The strut number of Z-shaped stent-grafts has a significant influence on biomechanical properties of the stent-grafts.Especially the Z-shaped stent-graft with relatively fewer strut numbers shows an advantage in evaluating radial support performance and flexibility such as the bending torque and the cross-section deflection,and is more suitable to be anchored and fit to the tortuous blood vessel,which will play a positive role in decreasing the complication such as endoleaks and thrombosis.The research findings can guide structure design of Z-shaped stent-grafts and optimal selection for surgery.

Establishment and identification of pancreatic stem cell strain derived from islets of Kunming mice under feeder layer conditions / 中国组织工程研究

BACKGROUND:Until now, little has been reported on establishment of pancreatic stem cell strains and lines,and the purification of pancreatic stem cells is difficult since the cell line establish rate is low.OBJECTIVE:To explore a more rational and effective technique of in vitro separation and continuous passage of pancreatic stem cells, with the hope to establish cell strains and even cell lines and to lay the foundation for the follow-up study of pancreatic stem cells in the treatment of diabetes.METHODS:Firstly, Percoll discontinuous density gradient centrifugation method was applied to separate the mouse pancreatic endocrine portion from the exocrine portion, then to obtain cell strains with highly proliferative ability and low differentiation from pancreatic endocrine portion-the islet. We used mouse embryonic fibroblasts treated with mitomycin C as a feeder layer, for in vitro continuous culture of islet-derived pancreatic stem cells under feeder layer conditions until they were transferred to the 30th passage to establish cell lines. Then pancreatic stem cell line derived from pancreatic islet was detected and identified by a series of tests including growth characteristic test, morphological observation, related molecular marker identification and differentiation characteristic identification.RESULTS AND CONCLUSION:In the continuous process of passage, pancreatic stem cells showed active proliferative ability, and maintained the typical morphological characteristics of stem cells and expression of pancreatic stem cell marker-Nestin. After induction, pancreatic stem cells showed insulin gene expression,reflecting their differentiation potential. Therefore, under the condition of feeder layer, the pancreatic stem cell line derived from Kunming mice was successfully established and the related identification was completed,which lays the foundation for the following research.