To clone and express 27 kDa cysteine protease (CP) gene of Spirometra erinacei plerocercoid,and analyze the biology characteristics,a total of RNA was extracted from the plerocercoids and reversely transcribed into cDNA.The 27kDaCP gene was amplified by PCR and cloned into pM-19T vector for sequencing.The accurate sequence was subcloned into the expression vector pET-28a (+).The recombinant plasmid was transformed into Transetta (DE3) and the expression protein induced by IPTG.The recombinant protein was purified by Ni2 + affinity chromatography,and analyzed by SDS-PAGE and Western blotting.The 27 kDa CP gene and its expression protein were predicted and analyzed by bioinformatics analysis tools such as NCBI and ExPASy.Results showed that the ORF length of 27 kDa CP gene sequence was 1 011 bp,and the removed signal peptide sequence was 954 bp with the submission number of ANA52569 in GenBank.The whole sequence of 27 kDa CP (Mr 35 669.9,pI 5.92) was 317 amino acids conferred from cDNA,which belongs to the Peptidase_C39_like superfamily.The pET-28a (+)-27kDa-CP was expressed under the induction of IPTG.Western blotting analysis showed that the purified protein reach expectancy,and had better response with positive serum of Spirometra erinacei plerocercoid infection.In conclusion,the 27 kDa CP gene of Spirometra erinacei plerocercoid is successfully cloned and expressed and knowing coded sequences and bioinformatic.

Objective To investigate the effect of intravenously transplanted bone marrow mononuclear cells (BMMNCs) on brain injury and inflammation after intracerebral hemorrhage(ICH) in rats.Methods Experimnental ICH models were performed by stereotaxic injection collagenase Ⅳ into caudate putamen,rats that underwent ICH were randomly divided into four groups:sham operation group,ICH group,PBS group,BMMNC-treated group.The BMMNCs were injected intravenously into rats after ICH.The neurobehavioral function was evaluated on days 1,3,7,14 by the modified neurological severity score,and the brain edema was examined by wet-dry weighting method on day 3 after cell transplantation.Immumofluorecence staining was used to identify the number of activation of microglia and infiltration of neutrophils in the brain after ICH.Results The neurological score in BMMNCtreated group on days 7,14 was significantly improved compared with those in ICH group and PBS group(P＜O.05).Compared to the ICH group ((81.09 ± 0.83) ％) and PBS group ((80.99 ± 0.79) ％),there was a significant decreasc in thc brain water content in BMMNC-treated group((78.62±0.97) ％) (P＜0.05).The number of activation of microglia and infiltration of neutrophils were both significantly lower in BMMNC-treated group ((55.8±22.1)/mm2,(49.6± 12.9)/mm2) compared to ICH group and PBS group (respectively (125.0 ± 20.7) /mm2,(86.8±13.6/mm2))(P＜0.01).Conclusion Administration of BMMNCs can significantly reduce edema and improve neurologic function by inhibiting the activation of microglia and infiltration of neutrophils.

Objective To develop a portbable incubator for on-site cultivation of microorganisms in foods and drinking water.Methods Cultivation temperature was set as required by the temperature for various microorganisms and PID was controlled via the single chip microcomputer and configuration screen .Then, the framework of the incubator was designed and assembled.Finally, the cultivation effect was tested .Results The incubator was compact and portable .The deviation of the temperature was in the range of 1℃.The hold time of self-contained power could exceed 8 h.In addition, the cultivation effect of our fabricated incubator was not significantly different from that of the commercial electro-heating standing-temperature cultivator used in laboratories .Conclusion The incubator is suitable for on-site detection of microorganisms in foods and drinking water , which is significant for spotting and removing the hidden dangers from microorganism contaminations in foods and drinking water in order to protect the health of soldiers .

bjective　To analyze the drug resistance screening status and drug resistance influencing factors of high-risk groups of drug-resistant pulmonary tuberculosis in Qingdao, and to understand the inclusion of rifampicin patients in treatment, so as to provide a reference for the prevention and treatment of drug-resistant pulmonary tuberculosis. Methods　The medical records of 726 cases of drug-resistant pulmonary tuberculosis among high-risk populations registered in Qingdao from 2018 to 2022 were obtained from the National Health Insurance Information System of the China Center for Disease Control and Prevention. The drug resistance to five anti-tuberculosis drugs, namely isoniazid (INH), rifampicin (RFP), ethambutol (EMB), levofloxacin (Lfx), and amikacin (Am), in the high-risk populations of drug-resistant pulmonary tuberculosis was analyzed. Univariate and multivariate logistic regression were used toidentify factors influencing rifampicin resistance, and the detection and inclusion of treatment for rifampicin-resistant patients were evaluated. Results　Of the 726 subjects, 278 were drug-resistant, with a total drug resistance rate of 38.29%. The drug resistance for the five anti-tuberculosis drugs in descending order was: INH 25.90%(188/726), RFP 22.87%(166/726), Lfx 14.19%(103/726), EMB 11.29%(82/726), Am 2.48%(18/726). Analysis of the drug resistance spectrum showed that among those resistant to one drug, RFP was most common, accounting for 13.67% (38/278); among those resistant to two drugs, INH+RFP was predominant, accounting for 15.83% (44/278); among those resistant to three drugs, INH+RFP+Lfx was most frequent, at 7.19% (22/278); and among those resistant to four drugs, INH+RFP+EMB+Lfx was highest, at 6.12% (17/278). Multivariate logistic regression analysis of rifampicin resistance showed that compared with patients under 25 years of age, the risk of developing rifampicin resistance was lower in the groups aged 45 to under 65 and those aged 65 and above (OR=0.356, 95%CI: 0.181-0.700; OR=0.352, 95%CI: 0.170-0.729). Compared with migrant patients in other provinces, local patients from within the same county or district had a lower risk of developing rifampicin resistance (OR=0.599, 95%CI:0.383-0.962). Compared with patients who were smear-positive at the end of the second month of initial treatment, the risk of developing rifampicin resistance was higher in patients with relapse/return, failure of retreatment/chronic, and other categories of patients (OR=9.380, 95%CI:3.717-23.671;OR=25.749, 95%CI:8.037-82.490; OR=36.651, 95%CI:8.438-159.201). Conclusions　The situation of drug-resistant pulmonary tuberculosis in Qingdao cannot be ignored. Individuals under 25 years old, migrants from other provinces, and patients with relapse/return, failure of retreatment/chronic, and other categories are significant risk factors for developing rifampicin resistance in the high-risk groups of drug-resistant pulmonary tuberculosis.

This article aims to summarize the significance of the establishment of human milk banks, the status of human milk banks in mainland China, analyze the relevant factors that affect the development of human milk banks, and propose corresponding countermeasures to provide a reference for improving the status of human milk banks in mainland China and promoting sustainable development.

Objective: To investigate the epidemic status and influencing factors of colorectal polyps among urban residents in Lanxi. Methods: A representative urban community was selected from Lanxi. A total of 935 local residents aged 18-80 years received questionnaire survey,physical examination,laboratory tests and colonoscopy. Multivariate Logistic regression analysis was applied to exam the potential influencing factors for colorectal polyps. Results : Eventually 880 residents were involved into the statistical analysis. Colorectal polyps were detected in 167 participants with a crude prevalence of 18.98% and adjusted prevalence for age and gender of 13.28%. The prevalence of colorectal polyps in men（28.70%）was significantly higher than that in women（12.92%,P<0.05）. The results of logistic regression analysis showed that age（OR=1.038,95%CI：1.018-1.057）,males（OR=1.919,95%CI：1.303-2.826）,the middle school level of education（OR=2.443,95%CI：1.507-3.961）and constitution of Yin-Asthenia（OR=0.203,95%CI：0.057-0.720）were influencing factors for colorectal polyps;among male participants,hypertension（OR=1.721,95%CI：1.018-2.908）was a risk factor for colorectal polyps;among female participants,age（OR=1.076,95%CI：1.042-1.111）,BMI（OR=1.099,95%CI：1.001-1.207）,the middle school level of education（OR= 3.507,95%CI：1.794-6.856）and constitution of Yin-Asthenia（OR=0.160,95%CI：0.033-0.770）were influencing factors for colorectal polyps. Conclusion Age,sex,education level and constitution of Yin-Asthenia were associated with the risk of colorectal polyps among residents in Lanxi;hypertension was associated with the risk of colorectal polyps in males,while age,BMI,education level and constitution of Yin-Asthenia were associated with the risk of colorectal polyps in females.

BACKGROUND:With the application and development of 3D printing technology in medicine,orthopedic internal fixation surgery has become precise and individualized.The equal-scale fracture model obtained by 3D printing technology was simulated and planned before surgery,realizing the leap from traditional 2D images to more vivid and detailed three-dimensional objects.It allows the surgeon to understand the fracture type in advance and rehearse the reduction sequence,so as to realize the individualized implementation of fracture surgery,optimize the surgical process,bring better postoperative recovery and less surgical complications. OBJECTIVE:To compare clinical efficacy of three 3D printed models combined with computerized virtual repositioning technology to assist incision reduction bone plate internal fixation and traditional incision reduction bone plate internal fixation in the treatment of Robinson II B2 clavicle fracture. METHODS:Eighty patients with Robinson II B2 clavicle fracture were randomly divided into trial group(n=40)and control group(n=40).In the trial group,three kinds of 3D printing models(affected clavicle fracture model,computer simulation clavicle fracture reduction model,clavicle mirror model of healthy side)combined with computer virtual reduction technology were used for preoperative in vitro surgery rehearsal.Finally,3D printing was used for clavicle mirror model of healthy side to advance bending and select bone plates for internal fixation.In the control group,open reduction plate internal fixation was applied.The time from admission to surgery,intraoperative blood loss,operation time,frequency of fluoroscopy,number of bends of the bone plate,fracture healing time,complications,and visual analog scale score and Constant score before and after surgery were compared between the two groups. RESULTS AND CONCLUSION:The time from admission to operation in the trial group was greater than that in the control group(P<0.05).Operation time,intraoperative fluoroscopy frequency and bending times of the bone plate in the trial group were lower than those in the control group(P<0.05).The trial group had faster fracture healing and fewer complications(P<0.05).There was no significant difference in intraoperative blood loss between the two groups(P>0.05).Constant score of the two groups had an increasing trend with time(F=613.50,P<0.001),but the difference between the groups was not statistically significant(F=0.08,P=0.78),and there was no interaction between the measurement times and the group assignment(F=0.27,P=0.66).The visual analog scale score decreased with time(F=1 149.55,P<0.001),but there was no significant difference between groups(F=0.02,P=0.88),and there was no interaction between the number of measurements and the group assignment(F=1.02,P=0.36).The results show that the use of 3D printed model combined with computer virtual reduction technology for preoperative rehearsal can shorten the operation time,reduce the number of intraoperative fluoroscopy frequency and the times of bone plate bending,and have the advantages of faster fracture healing,fewer complications,and similar functional recovery to the traditional incision reduction bone plate internal fixation.

Objective To investigate the effect of transplantation of bone marrow mononuclear cells (BMMNCs) on neuron-like cell differentiation and glial cell-derived neurotrophie factor (GDNF) expression in rats with intracerebral hemorrhage (ICH).Methods ICH was induced by stereotactic injection of type Ⅳ collagenase into the striatum of rats,and the successful models were randomly divided into model group (ICH group) and BMMNCs transplantation group (BMMNCs group); BMMNCs group was injected with 1 mL cell suspension containing 3 ×107 cells marked with 5-Bromo-2-deoxyuridine (Brdu).The neurological functions of the rats were observed by modified neurological severity scale (mNSS) test 1,7,14 and 28 days after the injection; the GDNF expression in cerebral tissue was observed by Western blotting 1,7,14 days after the injection; double immunofluorescent labeling was utilized to trace the differentiation of BMMNCs in the rat brains 28 days after the injection.Results As compared with those in the ICH group,the mNSS scores in the BMMNCs group were significantly lower (P＜0.05) and the GDNF expression was significantly increased at all time points (P＜0.05); Brdu and neuron specific nuclear protein (NeuN) double-labeled positive cells were found in the hemorrhage focus in the BMMNCs group 28 d after injection.Conclusion BMMNCs transplantation can improve the neural behavioral function,increase the GDNF expression,and induce BMMNCs differentiating into neuron-like cells.

Compared with other drug loading methods,PLGA drug-loaded microspheres have the advantages of sustained and con-trolled release ability,biodegradability,drug release stability,and targeted delivery function,which have become a research hotspot in pharmaceutics.With the improvement of the preparation technology and the quality of microspheres,they have been gradually promoted in clinical practice,and have achieved remarkable results in the treatment of tumors,osteoarthritis,diabetes,eye diseases and so on.However,there is still a lack of systematic review on the application of PLGA microspheres in stomatology.This article aims to intro-duce the clinical application characteristics of PLGA microspheres,and review their application prospect in oral and maxillofacial tumors,temporomandibular joint diseases,periodontitis,caries and alveolar bone defects,so as to provide reference for their applica-tion in stomatology.

Objective:To investigate the clinical, pathologic and radiologic features and molecular alterations in patients with primary cardiac leiomyosarcoma (PCLMS).Methods:Five cases of PCLMS were collected in Beijing Anzhen Hospital from January 2016 to December 2020. The clinical, pathologic and radiologic data, and molecular alterations were analyzed, and the patients were followed up.Results:All five patients were female, and had no history of leiomyosarcoma in other parts of the body. The age of patients ranged from 37 to 62 years (median 47 years). The main clinical symptoms were chest pain and dyspnea, one also presented with palpitation and lower limb weakness and one with dizziness. Two tumors were located in the left atrium, two in the right atrium, and one in the right ventricle, and they maximal diameter ranged from 2.5 to 14.0 cm (mean 6.2 cm). The neoplasms presented as medium-echo masses with a broad base in the echocardiography, and as a low-density, solid mass when detected by contrast-enhanced CT. Histologically, two tumors were well-differentiated and three were moderately and poorly differentiated, and two included extensive, loose myxoid stroma. Immunohistochemical staining showed that PCLMS was positive for SMA, desmin, MDM2, and epidermal growth factor receptor. Fluorescence in situ hybridization showed ALK gene rearrangement in two cases, and COL1A1-PDGFB fusion in three cases. All cases received surgical excision and two cases received chemotherapy. Three patients died within 0-11 months (mean survival of 7.7 months) and two patients were alive.Conclusions:PCLMS is a malignant tumor with a high recurrence rate and poor prognosis. These cases may provide useful information to improve the diagnosis and management of PCLMS.