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 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 .

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.

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.

AIM: To investigate the protective effect and mechanism of Danlou tablet on retinal ischemia-reperfusion injury(RIRI)in mice.METHODS: A total of 40 ApoE-/- mice were fed with high fat diet for 6 wk, and the RIRI model was established by anterior chamber infusion of pressurized saline. The mice were divided into control group(normal saline for 8 wk), RIRI model group(normal saline for 8 wk), and low-, medium-, and high-dose Danlou tablets groups [1, 2, and 4 g/(kg·d), respectively, for 8 wk]. The morphological changes of retina were observed by hematoxylin-eosin(HE)staining, retinal cell apoptosis was detected by terminal-deoxynucleoitidyl transferase mediated Nick-End Labeling(TUNEL)staining. The Western-blot assay was used to detect the expression of retinal tissue sample Kelch-like ech-associated protein 1(Keap1), nuclear factor E2 related factor 2(Nrf2), heme oxygenase 1(HO-1), and superoxide dismutase(Sod2)proteins.RESULTS: Compared with the control group, the mouse retina was atrophic with thinning thickness and increasing cell apoptosis, down-regulation of Sod2 protein expression, and up-regulation of Keap1 protein expression in the RIRI model group(all P<0.01). Compared with the RIRI model group, the retinal thickness increased in the medium- and high-dose of Danlou tablets groups(all P<0.01), and the cell apoptosis of retina decreased in the low-, medium- and high-dose of Danlou tablets groups(all P<0.05). There were no significant differences in the expression of Keap1 and HO-1 proteins of mouse retina tissue in the low-dose of Danlou tablets group(P>0.05). The expression of Sod2, Nrf2 and HO-1 proteins up regulated, and the expression of Keap1 protein down regulated in the medium- and high-dose of Danlou tablets groups(all P<0.05).CONCLUSION: Danlou tablet can alleviate RIRI-induced atrophy and thinning of retina and retinal cell apoptosis by regulating Keap1-Nrf2/HO-1 signal pathway and reducing oxidative stress.

The main fundus changes of pathologic myopia (PM) are posterior staphyloma (PS) and myopic maculopathy (MM), which includes myopic atrophy maculopathy (MAM), myopic tractional maculopathy (MTM), myopic neovascular maculopathy (MNM) and so on. The clinical manifestations of PM-related fundus lesions are complex, and the classification of PM has been a research hotspot in recent years. The proposal of each classification shows an increasing understanding of PM, and each classification has its advantages but also imperfections. For MM, it is recommended to refine the MTM classification based on the ATN classification and adjust it according to the internal correlation between MAM and MNM. The rapid development of modern imaging technology will promote the continuous update of the classification, and its further improvement will also help to understand the development process of PM, which has important clinical value in preventing its occurrence and progression.

Developing new therapeutic agents for cancer immunotherapy is highly demanding due to the low response ratio of PD-1/PD-L1 blockade in cancer patients. Here, we discovered that the novel immune checkpoint VISTA is highly expressed on a variety of tumor-infiltrating immune cells, especially myeloid derived suppressor cells (MDSCs) and CD8⁺ T cells. Then, peptide C1 with binding affinity to VISTA was developed by phage displayed bio-panning technique, and its mutant peptide VS3 was obtained by molecular docking based mutation. Peptide VS3 could bind VISTA with high affinity and block its interaction with ligand PSGL-1 under acidic condition, and elicit anti-tumor activity in vivo. The peptide DVS3-Pal was further designed by d-amino acid substitution and fatty acid modification, which exhibited strong proteolytic stability and significant anti-tumor activity through enhancing CD8⁺ T cell function and decreasing MDSCs infiltration. This is the first study to develop peptides to block VISTA/PSGL-1 interaction, which could act as promising candidates for cancer immunotherapy.