Objective To develop a method for rapidly identifying Clostridioides difficile(C.difficile)and de-termining high-producing toxin strains,conduct clinical evaluation.Methods The loop-mediated isothermal amplifi-cation(LAMP)method was used to identify C.difficile based on the tcdC,tcdA,and tcdB genes.The sensitivi-ty,specificity,and overall consistency of the detection method were evaluated.Results Feces specimens from 499 hospitalized patients suspected of C.difficile-associated diarrhea were detected,with C.difficile detection rate of 12.8％(64/499),out of which the detection rate of toxin-producing C.difficile was 10.8％(54/499).The sensi-tivity,specificity,positive predictive value,and negative predictive value of the detection method for tcdA were 87.2％,98.9％,89.1％,and 98.6％,respectively,and 88.2％,99.6％,90.0％,and 98.73％for tcdB,respec-tively.The total toxin levels of different strains were different,but the average toxin production level of A+B+strains(1.79 μg/mL)was higher than those of A-B+strains(0.72 μg/mL)and A-B-strains(＜0.10 μg/mL).Conclusion The portable high-throughput LAMP detection method can rapidly and efficiently identify C.difficile and determine high-producing toxin strains.

Thoracic anesthesia with spontaneous respiration represents a form of precision anesthesia meticulously customized to individual patients.Considering the more stringent requirements this anesthesia approach imposes on the regulation of respiratory function,the writing group of the"Consensus of Experts on the Management of Thoracic Anesthesia with Spontaneous Respiration"has formulated elaborate guidelines regarding indications and contraindications,preoperative evaluation,anesthesia implementation,common complications,and treatment strategies.This was accomplished by referencing relevant domestic and international literature and integrating it with actual clinical requirements.The objective is to standardize the rational application of this anesthesia method.

Objective To develop a method for rapidly identifying Clostridioides difficile(C.difficile)and de-termining high-producing toxin strains,conduct clinical evaluation.Methods The loop-mediated isothermal amplifi-cation(LAMP)method was used to identify C.difficile based on the tcdC,tcdA,and tcdB genes.The sensitivi-ty,specificity,and overall consistency of the detection method were evaluated.Results Feces specimens from 499 hospitalized patients suspected of C.difficile-associated diarrhea were detected,with C.difficile detection rate of 12.8％(64/499),out of which the detection rate of toxin-producing C.difficile was 10.8％(54/499).The sensi-tivity,specificity,positive predictive value,and negative predictive value of the detection method for tcdA were 87.2％,98.9％,89.1％,and 98.6％,respectively,and 88.2％,99.6％,90.0％,and 98.73％for tcdB,respec-tively.The total toxin levels of different strains were different,but the average toxin production level of A+B+strains(1.79 μg/mL)was higher than those of A-B+strains(0.72 μg/mL)and A-B-strains(＜0.10 μg/mL).Conclusion The portable high-throughput LAMP detection method can rapidly and efficiently identify C.difficile and determine high-producing toxin strains.

Thoracic anesthesia with spontaneous respiration represents a form of precision anesthesia meticulously customized to individual patients.Considering the more stringent requirements this anesthesia approach imposes on the regulation of respiratory function,the writing group of the"Consensus of Experts on the Management of Thoracic Anesthesia with Spontaneous Respiration"has formulated elaborate guidelines regarding indications and contraindications,preoperative evaluation,anesthesia implementation,common complications,and treatment strategies.This was accomplished by referencing relevant domestic and international literature and integrating it with actual clinical requirements.The objective is to standardize the rational application of this anesthesia method.

Objective:To explore the correlation of distribution pattern of TCM pathological evidence of hypertension with the related indexes of cardiopulmonary function.Methods:From January 2019 to October 2021, the relevant data of 149 patients with grade 3 hypertension who underwent CPET examination in the Department of Cardiology, Qingdao Hospital of Traditional Chinese Medicine were retrospectively collected, and the pathogenic factors were determined according to the relevant content of "Dialectic of Evidence". The Chi-square test was used to analyze the difference of cardiopulmonary function between the pathological factors, and the Spearman correlation analysis was used to explore the correlation between pathological factors and cardiopulmonary function.Results:The pathogenic factors of 149 hypertensive patients from high to low, are as follows were phlegm, yin deficiency, dampness, yang hyperactivity, blood deficiency, blood stasis, qi deficiency, yang deficiency, qi stagnation, heat, food accumulation, moving wind, pus. There was no statistical significance among the pathological factors ( P>0.05). Spearman correlation analysis showed that Yin deficiency syndrome was negatively correlated with Peak O 2 pulse ( r=-0.213, P=0.009), blood stasis was negatively correlated with Peak VO 2 ( r=-0.170, P=0.039) and Peak O 2 pulse ( r=-0.192, P=0.019), and phlegm syndrome hormone was negatively correlated with anaerobic threshold (AT) ( r=-0.192, P=0.019). Conclusions:TCM pathogenic factors in hypertension mainly focus on phlegm, Yin deficiency, dampness and yang hyperactivity. There is a certain correlation between pathogenic factors and cardiopulmonary function-related indicators, which can provide reference for the clinical treatment of hypertension in TCM.

Objective To explore changes of invasion capability of cancer stem cells loading miR30a derived from enlarging and proliferative axillary lymph node of patients with breast cancer in nude mice.Methods MiR30a oligonucleotide fragment loaded by adenovirus vector was transfected into breast cancer stem cells isolated from enlarging and proliferative axillary lymphnodes of patients with breast cancer,MDA-MB-231 cell lines as the control.The cancer cells were injected into axillary subcutaneous fat of the nude mice for several weeks,and then,the expression of Vimentin or N-Cadherin in tumor tissues of each group of the mice was determined by immunohistochemistry and western blot.Results The average transfection rate of cancer stem cells loaded by miR30a was 62.5％,while it was 78.2％ for MDA-MB-231.The tumor volume was larger in adenovirus vector groups or in control groups of nude mice than in experimental groups induced by mir30a.Vimentin or N-cadherin in tumor tissues was significantly downexpressed in experimental groups with mir30a ((13.1±1.7)％,(15.3％±2.1)％)compared with that in adenovirus vector groups or in control groups((21.1±1.4)％,(25.3±1.6)％,P＜0.05),respectively.The difference between adenovirus vector groups and in control groups had no significant difference (P＞0.05).After inoculated for 6 weeks,except the subcutaneous plantations,no distant metastasis in nude mice was found.MDA-MB-231 cell lines groups had similar results.Conclusion The proliferative and invasive capability of cancer stem cells can be inhibited by miR30a,suggesting a new therapy for breast cancer.

Objective To compare the biomechanical characteristics of new assembly of locking compression plate (NALCP) and locking compression plate (LCP) in internal fixation of femoral shaft comminuted fractures.Methods The preparation of a femoral shaft wedge fracture model (AO type 32-C2.1),six pairs of (12) femoral specimens were collected and divided into two groups randomly,with six in each group.The Group A was made up of the new assembly of locking compression plate fixation model (NALCP),and Group B the locking compression plate fixation model (LCP).The biomechanical properties of steel plates in two groups were tested by axial loading and torsional loading tests.The relative maximum displacement of fracture blocks in two groups on the X,Y and Z axes (the coronal axis was set as X axis,through the medial and lateral femur;the transverse axis was set as Z axis,through the femoral intercondylar fossa,perpendicular to the X axis;the sagittal axis was set as Y axis,perpendicular to the X and Z axis),the maximum strain,and the average strain of the steel plate were recorded.Strain distribution nephogram was produced,and the axial loading fatigue test results of Group A were recorded.Results Axial loading test:the relative maximum displacement of fracture in Group A on X,Y and Z axis were smaller than those in Group B (P ＜0.05 or 0.01);the main strain of Group A was greater than that of Group B (P ＜0.01);there was no significant difference in the average strain between Group A and Group B (P ＞ 0.05).Torsional loading test:The relative maximum displacement of fracture in Group A on X and Z axis was smaller than that of Group B (P ＜ 0.01);there was no statistically significant difference between the two groups on the Y axis (P ＞ 0.05);the main strain of plate in Group A was greater than that in Group B (P ＜ 0.01);there was no statistically significant difference in the average strain between Group A and Group B (P ＞ 0.05).There was no obvious difference in strain distribution between the two groups.In Group A,the fatigue test of axial cyclic loading was performed for 1 million times,and the NALCP was intact without deformation,loosening,or rupture.Conclusion NALCP can provide strong mechanical stability for comminuted femoral fracture.The design of bridge steel plate is reasonable,which can effectively avoid stress concentration,reduce the stress shielding of steel plate,and facilitate bone healing.

Objective To compare the stress and its distribution between our self-designed new assembly of locking compression plate (NALCP) of low elastic modulus versus conventional locking compression plate (LCP) in fixation of femoral comminuted fractures.Methods Six pairs of cadaveric femur were used to create models of middle femoral comminuted fracture.The femoral fracture models were fixated respectively by NALCP of Ti2448 with low elastic modulus (E =30 Gpa) (NALCP group) and conventional LCP of Ti-6Al-4V with high elastic modulus (E =110 Gpa) (LCP group).Axial and torsion loads were applied on the models in the 2 groups to simulate those on one leg when a person slowly walks.The relative maximum displacements on the X,Y and Z axes of fracture fragments,and the maximum and average strains of the plate were recorded in the 2 groups.Nephograms of strain distribution were made for the 2 groups.The results of fatigue test under axial loads were recorded for NALCP group.Results Both the axial and torsion loading tests showed significantly larger principal and average strains in NACLP group than in LCP group (P ＜ 0.05).However,there were no significant differences between the 2 groups in the relative maximum displacements of fracture fragments on X,Y or Z axis (P ＞ 0.05).The plate strain nephograms for the 2 groups showed consistent strain distributions.The plates in NALCP group survived 1,000,000 fatigue tests under axial loads,without any deformation,loosening or breakage.Conclusion As our NALCP of low elastic modulus may be better in stress transmission and distribution,it can effectively reduce the effect of stress-shielding and promote bone healing.

Objectives To detect the expression level of miR-155 in tissues of colon cancer,and to investigate its influence on the occurrence,development and metastasis of colon cancer.Methods 69 cases with primary colon cancer treated with surgical resection in our hospital were chosen.Tissue samples of the tumor and tissue in the side of tumor(control)were collected and the expression level of miR-155 in the samples was detected by fluorescence quantitative polymerase chain reaction.The results were statically analyzed.Results The expression level of miR-155 was 0.471±0.100,0.238 ±0.108,0.719 ± 0.135 in three groups of colon cancer patients with tumor sizes ≥ 5 cm,high differentiation of tumor,and occurrence of lymphatic metastasis,respectively,and 0.623 ± 0.134,0.142 ±0.094,0.390±0.121 in their controls.And the level of miR-155 was 0.395±0.101 in patients with T1 + T2 tumor local infiltration,0.466±0.128 in T3 + T4 local infiltration,0.404±0.108 in Ⅰ + Ⅱ stage of TNM and 0.457±0.119 in Ⅲ± Ⅳ stage.The miR-155 was relatively higherly expressed in patients with tumor sizes≥5 cm,lower degree of differentiation,higher tumor infiltration degree,Ⅲ and Ⅳ stage of TNM,(t=3.002,12.531,20.425,2.576,1.908,P ＜ 0.05).Conclusions The expression level of miR-155 is correlated with tumor sizes,differentiation degrees,tumor infiltrating degrees,TNM and lymphatic metastasis,which could be used as diagnostic indexes for occurrence,development and metastasis of colon cancer.

Research-oriented study is a comprehensive learning method focusing on the experimental process deriving from a main issue.Given our clinical teaching experience in years,we incorporated the research-oriented studies into the undergraduate Clinical Medicine course for grade 2009,integrating the trait of the clinical skills education and our local professional training objectives.In the studies,students selected their own topic,designed and did their experiments,searched for the information,processed and analyzed the experiment result and finally completed the thesis.Practice showed that after the strict implementation of research learning plan,students' learning interest,innovative thinking,practical skills,research ability and cooperation spirit all significantly enhanced.