Objective: To adjust the distance between SD sequence and ATG in the same expressive plasmid pLX1 to enhance expression of heterologous bFGF gene in E. coli. Methods: Adjusting the different distance between SD sequence and ATG by Klenow and Mung Bean Nuclease. SDS PAGE and Western blot showed the expressed protein bFGF in E.coli. bFGF proteins were purified by HPHIC, HPGFC and HAC. Biological activity was examined by MTT. Results: Recombinant plasmids pLX2, pLX3 were obtained and the expressive levels were 8.03%, 9.9% respectively. Also the purified bFGF was obtained by HPHIC, HPGFC, HAC and its ED 50 was 2.29 ng/ml. Conclusion: Increasing the bFGF gene dosage by adjusting the distance between SD sequence and ATG could increase the expression level of a desired protein.

Objective To identify the relationship among serum 25-hydroxyvitamin D[25(OH)D] level, chronic inflammation state and malnutrition in maintenance hemodialysis (MHD) patients. Methods The serum 25 (OH)D level of 119 patients on MHD was detected. All the patients were divided into three groups according to the serum 25(OH)D level. Vitamin D deficiency: 25(OH)D ≤ 15 ng/mL, vitamin D insufficiency: 15 ng/mL <25(OH)D ≤ 30 ng/mL, vitamin D normal: 25(OH)D > 30 ng/mL. Inflammatory factor and nutritive index were compared among the three groups. Result The average of serum 25 (OH)D level was 3.4~45.3 ng/mL (22.5~14.8 ng/mL), and the prevalence of 25 (OH)D deficiency and insufficiency was 88.2%. Significant differences existed in age, inflammatory factor (IL-6, TNF-α, CRP) and nutritive index (Alb, PA, SGA) between 25(OH)D deficiency, insufficiency groups and normal group (P<0.05). Conclusion The serum 25(OH)D level is possibly related to the malnutrition in MHD patients. It may inhibit chronic inflammation state , in which it can promote state of nutrition of MHD patients.

Objective:To explore the curative effect of acupuncture combined with huanglian wendan decoction on the mild cognitive impairment after stroke.Methods:100 patients with mild cognitive impairment after stroke were enrolled in our hospital from June 2013 to August 2016 and randomly divided into two groups.The control group (n=50) accepted nimodipine,and the study group (n=50)accepted huanglian wendan decoction combined with acupuncture treatment based on the control group.The MMSE and MSSS scores were evaluated and compared,and the improvement of cognitive dysfunction and nervous dysfunction,quality of life were compared and analyzed.Results:After treatment,the MMSE,ADL scores of patients were remarkably decreased compared with those before treatment (P＜0.05),and the decreasing degree of study group was significantly higher than those of the control group (P＜0.05).The MSSS,MBI scores of both groups were significantly increased compared with those before treatment (P＜0.05),and the increasing degree of study group was significantly higher than those of the control group (P＜0.05).The effective rates of improvement in cognitive dysfunction and nervous dysfunction,quality of life of study group were significantly higher than those of the control group (P＜0.05).Conclusions:Huanglian wendan decoction combined with acupuncture treatment was effective for the mild cognitive impairment after stroke and could improve the quality of life.

Objective To compare complications incidence rates of 8-core and 12-core prostate biopsies guided with transrectal ultrasound retrospectively,and to study intervention ways to decrease complications.Methods The data of 260 consecutive patients undergoing first-time transrectal ultrasound-guided biopsy were analyzed.132 patients underwent 8-core biopsy and 128 patients underwent 12-core biopsy.Results In 8-core group,there were 7 cases of infections,15 cases of gross hematuria,7 cases of rectal bleeding and 6 cases of acute urinary retention,and in 12-core group,5 cases of infections,19 cases of gross hematuria,9 cases of rectal bleeding and 7 cases of acute urinary retention in 12-core group.There are no statistical differences of complications incidence rate between two groups (P ＞ 0.05).when prostate volume ＞ 45 ml,detection rate of prostate cancer in 12-core group was significantly higher than that in 8-core group.Conclusions There are no statistical differences of complications incidence rate between 8-and 12-core prostate biopsy.Homologous measure must be taken to reduce complications.12-core prostate biopsy is recommended as prostate volume ＞ 45 ml.

To strengthen the pediatric residents' communication ability in rediatric residency standardized training is very important for improving quality of medical provider-patient relationship and prevent medical disputes. At present, in China's pediatric medical units at all levels of medical care for the hospitalized children groups there exist such characteristics as only child, parents doting over their children, excessive stress and fear in diagnosis and treatment and so on. From the analysis of current status, it can be seen that in clinical practice, pediatric physicians' doctor-patient communi-cation is affected by three aspects: children, parents, physicians , and there exist such problems as children's lack of coordination, parents' lack of medical care knowledge, residents' lack of clinical and communication experience. Therefore, it is necessary to strengthen the special training of pediatric residents' doctors and patients in the standardized training period, establish the feedback mechanism of communication between doctors and patients and. make targeted improvements in view of the weak link in the communication between doctors and patients, so as to improve the training program of the pediatric residents' doctor-patient communication ability.

Objective To research the functional polymer liposome-trapped gold nanoparticles as molecular probes used as CT contrast agents.Methods Gold nanoparticles were entrapped by ultrasonic micro-emulsion method using self-developed linoleic acid-modified dextran polymer liposome.Under the same concentrations,clinical imaging was performed with Iohexol as control for the analysis of CT value differences.Then,the particle suspensions at appropriate concentration(6 mg/mL)were intravenously injected into Balb/c nude mice for real-time observation of CT imaging.ResultsIn vitro results showed that nanoparticles at greater than 4 mg/mL could present much higher CT value than Iohexol at the same conditions,while in vivo studies indicated nanoparticles could clearly show different organ contrast with prolonged circulation time.Conclusion Compared to clinical contrast agent Iohexol,functional polymer liposome entrapped gold nanoparticles could possess a great application prospect to be used as a new type of molecular probes for CT enhanced imaging.

In order to meet the market demand for the fast and accurate genetic detection of African swine fever virus(ASFV),a new method for EIS-qPCR detection was established with a rapid,high sensitivity,and pollution prevention.Primers and probes for a duplex qPCR were designed based on the conserved region of ASFV virus p72 gene and the endogenous internal standard(EIS)cytb gene sequence in pigs.An anti-contamination system was established with uracil DNA N-gly-cosylase enzyme in the reaction system.The results showed that the method can finish the rapid qPCR detection of ASFV within 30 min with a minimal detection limit of 4.12 copies/μL.Moreo-ver,the method only detected the ASFV p72 gene,and no amplifications of classical swine fever vi-rus(CSFV),pseudorabies virus(PRV),porcine parvo virus(PPV),porcine reproductive and re-spiratory syndrome virus(PRRSV)and porcine circovirus type 2(PCV2)were observed.Repeti-tive results showed a coefficient of variation below 2％.With strong anti-pollution capacity,the method can effectively eliminate false-positive amplification caused by low-dose aerosol pollution.Detection results of 146 clinical samples showed a 100％consistence with the results of the com-mercial ASFV detection kit.Compared with similar technologies,the EIS-qPCR established in this study was faster,sensitive,and suitable for the rapid diagnosis of ASFV infection in the early stage,which provided the tool for the monitoring and precise prevention and control of African swine fever.

Objective:To investigate the value of chromosomes 7 and 8 polysomy in circulating tumor cells (CTCs) for the diagnosis of non-small cell lung cancer, and the correlation of CTCs with clinical pathological characteristics and epidermal growth factor receptor (EGFR) mutations in cancer tissue.Methods:Fifty-seven patients with non-small cell lung cancer and 21 patients with benign lung diseases were enrolled at Beijing Chaoyang Hospital, Capital Medical University, Beijing, China from November 2017 to October 2020. Negative enrichment combined with immunofluorescence in situ hybridization (imFISH) was used to identify CTCs polysomy on chromosomes 7 and 8. EGFR mutations in 56 lung cancer patients was detected using ARMS-PCR.Results:CTCs were detected in 93.0% (53/57) of non-small cell lung cancers and 28.6% (6/21) benign lung lesions. The difference between lung cancer patients and the control cohort was statistically significant ( P<0.01). Receive operator curve (ROC) analyses showed that, when the cut-off value was 1 cell/3.2 mL, Youden index had the highest sensitivity of 93.0% and specificity of 71.4% (AUC=0.906, 95% CI:0.833-0.980, P<0.01). The positive rate of CTCs in stage Ⅲ-Ⅳ cancers was significantly higher than that in stage Ⅰ-Ⅱ ( P=0.023). No significant correlation was observed between positive rate of CTCs or chromosome polysomy and age, gender, smoking status, pathologic types and EGFR mutation status. The number of CTCs in EGFR mutated group was higher than that in the non-mutated group (6.5±1.1 vs. 3.7±0.7, P=0.045). The detection rate for CTCs ≥5 in the EGFR mutated group was also higher than the EGFR non-mutated group (52.0% vs. 19.4%, P=0.010). Conclusion:Detection of CTCs with chromosomes 7 and 8 polysomy has potential value in auxiliary diagnosis of non‐small cell lung cancer, and the number of CTCs is correlated to TNM stage and EGFR gene mutation status.

Objective:To study the application effect of the Benner's theory stratified training based on the "Protect the World" platform for nurses specialized in neurological rehabilitation.Methods:A total of 70 nurses who underwent on-the-job training for neurorehabilitation nursing specialists in 2023 were divided into control group ( n=35) and observation group ( n=35) based on random odd and even numbers to receive different training methods. The control group received routine nursing training, while the observation group received Benner's theory stratified training based on the "Protect the World" platform. The theoretical and skill assessment scores, job competency, and satisfaction scores of two groups of nurses were compared before and after training. SPSS 25.0 was used for t-test and chi-square test. Results:Before training, there were no significant differences between the two groups of nurses in terms of theory scores [(88.35±4.41) vs. (89.43±4.07)] and skill assessment scores [(89.22±3.27) vs. (88.43±3.16)]. After training, the theoretical and skill assessment scores were significantly higher in the observation group as compared with the control group [(95.51±5.01) vs. (90.24±4.99) and (95.15±4.24) vs. (91.13±4.33), both P<0.05]. After training, the competency scores and total scores of education guidance, management function, diagnostic function, assistance role, and intervention treatment were significantly higher in the observation group than in the control group ( P<0.05). The satisfaction survey scores were significantly higher in the observation group than in the control group ( P<0.05). Conclusions:The Benner's theory stratified training based on the "Protect the World" platform can improve the theoretical and skill assessment scores of nurses specialized in neurological rehabilitation. This approach significantly boosts their overall professional competence and holds considerable potential for broader adoption.

In recent years, new orthopaedic implantable devices continue to emerge, which require higher requirements for technical evaluation. Animal study is an important part of the research and development process for the new orthopedic implantable devices, which provides relevant evidence for product design and stereotyping. By introducing the purpose of animal study, and the application of 3R principle (replacement, reduction, refinement) in this field, we summarize the concern on the animal study, in order to provide reference for the development and research of new orthopedic implantable devices and biomaterials. At the same time, the application of evidence-based research methods such as systematic review in the field is introduced, which provides new tools and approaches for the technical review and regulatory science.
Animals ; Orthopedics ; Biocompatible Materials ; Prostheses and Implants ; Research Design