Robust and efficient control of therapeutic gene expression is needed for timing and dosing of gene therapy drugs in clinical applications. Ribozyme riboswitch provides a promising building block for ligand-controlled gene-regulatory system, based on its property that exhibits tunable gene regulation, design modularity, and target specificity. Ribozyme riboswitch can be used in various gene delivery vectors. In recent years, there have been breakthroughs in extending ribozyme riboswitch's application from gene-expression control to cellular function and fate control. High throughput screening platforms were established, that allow not only rapid optimization of ribozyme riboswitch in a microbial host, but also straightforward transfer of selected devices exhibiting desired activities to mammalian cell lines in a predictable manner. Mathematical models were employed successfully to explore the performance of ribozyme riboswitch quantitively and its rational design predictably. However, to progress toward gene therapy relevant applications, both precision rational design of regulatory circuits and the biocompatibility of regulatory ligand are still of crucial importance.

BACKGROUND:Vertebral column resection is the frequently mentioned spinal orthopaedic concept. Due to the high requirement of the operation skil , difficulty and more complications, the previous studies have reported from different aspects, and many researchers have focused on the analysis of complications, that may be related with the procedure and manner. OBJECTIVE:To analyze the efficacy and complications of posterior vertebral column resection combined with titanium screw rod fixation for the treatment of severe spinal deformity. METHODS:We retrospectively analyzed 48 patients with severe spinal deformity who treated with posterior vertebral column resection and titanium screw rod fixation, with an average removal of 1.6 vertebral. The patients were fol owed-up for 15-64 months. The Cobb angle (coronal plane and sagittal plane) of the patients before treatment, after treatment and in the final fol ow-up was analyzed, and the relative complications of the surgery were analyzed. RESULTS AND CONCLUSION:The patients with spinal deformity were divided into five categories, included kyphoscoliosis (n=11), severe scoliosis (n=20), congenital spinal deformity (n=4), spherical kyphosis (n=3), and angular kyphosis (n=10). The average coronal plane deformity angle of the patients was corrected from 84° preoperation to 35° postoperation, with the total correction rate of 54%. The average sagittal plane deformity angle was corrected from 90° preoperation to 42° postoperation, and the sagittal plane Cobb angle was decreased for 48°. The mean operation time was 545 minutes (204-1 355 minutes), the intraoperative blood loss was 1 610 mL (50-8 244 mL), and the average blood loss was 65%. After treatment, 31 cases had complications, including 13 cases of intraoperative neurological dysfunction (observed through electrophysiological monitoring and wake), permanent neurological dysfunction did not occur after timely treatment. The posterior vertebral column resection and titanium screw rod fixation can obtain better effect in the treatment of severe spinal deformity, but the procedure has high complications and is difficult for operation.

Background and purpose:Due to the lack of cost-effective pre-treatment predictors for advanced cervical squamous cell carcinomas treated with concurrent chemoradiotherapy (CCRT), both baseline circulating CD4+CD25+CD127Low/- regulatory T cell (Treg) count and serum squamous cell carcinoma antigen (SCC-Ag) level were measured for this feasibility study. Methods: Peripheral blood samples were collected from 44 patients with stageⅡB-ⅣA cervical squamous carcinomas before CCRT. Flow cytometry immunophenotyping and enzyme-linked immunosorbent assay were used for circulating CD4+CD25+CD127Low/-Treg count and serum SCC-Ag level testing,respectively. Clinical and pathological characteristics were retrospectively reviewed to analyze the predictive value of the 2 indexes. Results:The baseline circulating CD4+CD25+CD127Low/-Treg count was lower in the patient group with positive treatment response than in the group with negative response [(8.78±2.80)%vs (10.95±2.56)%, P<0.05], and the serum SCC-Ag level showed no signiifcant difference between the 2 groups. No correlation was detected between these 2 markers (Spearman’rho=-0.093, P=0.540). Determined by plotting receiver operating characteristic curves, the best cut-off points were 9.76%for circulating CD4+CD25+CD127Low/-Treg count and 9.50 ng/mL for serum SCC-Ag level, respectively. Univariate analysis showed that pretherapeutic circulating CD4+CD25+CD127Low/-Treg count (OR=1.901, 95%CI:1.112-3.219, P=0.017), but not serum SCC-Ag level (OR=0.998, 95%CI:0.001-4.253, P=0.897), was predictive of clinical response to CCRT. Multivariate Logistic regression analysis revealed that pre-treatment CD4+CD25+CD127Low/-Treg count was an independent predictor for clinical response to CCRT (OR=3.115, 95%CI:1.253-7.742, P=0.014). Conclusion:Pretherapeutic circulating CD4+CD25+CD127Low/-Treg count is a feasible method to predict clinical response to CCRT in patients with advanced cervical squamous cell carcinomas.

Objective To construct a prokaryotic expression plasmid pET-28a(+) encoding the multiple transferable resistance C (mtrC) gene of N. gonorrhoeae and express it in E.coli DE3, in order to provide a model to study the pathogen's resistance mechanisms to antimicrobial hydrophobic agents. Methods The mtrC gene of N. gonorrhoeae was amplified by polymerase chain reaction from reference strains,cleaved with restriction endonuclease, and then cloned into the prokaryotic expression plasmid pET-28a (+) to construct the recombinant pET-mtrC. This was confirmed by cleavage of restriction endonuclease and DNA sequencing. The recombinant pET-mtrC was transformed into E.coli DE3 to express the protein MtrC with induction by IPTG. Results The mtrC gene in the recombinant pET-mtrC showed 99.5% homology with the reference sequence in GeneBank (U14993). A 48.5 kD fusion protein was identified by SDS-PAGE. Conclusions The successful construction of a prokaryotic plasmid encoding the mtrC gene of N. gonorrhoeae and its expression in E.coli may facilitate the development of a monoclonal antibody to the MtrC protein and help to investigate the mechanism of the mtr efflux system of N. gonorrhoeae.

Objective To measure the expressions of programmed death?1(PD?1)and programmed death ligand?1(PD?L1)on peripheral blood T lymphocytes of patients with condyloma acuminatum(CA), and to investigate their role in cellular immunity in these patients. Methods Peripheral blood samples were obtained from 30 patients with CA(CA group)and 20 healthy human controls (control group). Flow cytometry was conducted to detect the expressions of PD?1 and PD?L1 on the surfaces of peripheral blood CD4+and CD8+T lymphocytes, and to determine the counts of CD4+and CD8+T lymphocytes. Enzyme?linked immunosorbent assay(ELISA)was performed to measure the levels of serum interleukin?2(IL?2)and interferon?γ(IFN?γ). Statistical analyses were carried out to compare the above parameters between the two groups, and to assess the relationship of PD?1 and PD?L1 expressions with the counts of CD4+and CD8+T lymphocytes as well as with the serum levels of IL?2 and IFN?γ. Results There was a significant increase in the expression rates of PD?1 and PD?L1 on CD4+T lymphocytes(PD?1:9.48%± 3.31%vs. 7.12%± 2.16%, t=2.81, P<0.01;PD?L1:4.40%± 1.46%vs. 3.26%± 1.13%, t=3.16, P<0.01)and CD8+T lymphocytes(PD?1:12.52%± 3.17%vs. 9.95%± 2.17%, t=3.16, P<0.01;PD?L1:7.07%± 2.23%vs. 5.39%± 1.69%, t=2.88, P<0.01)in the CA group compared with the control group. Moreover, the CA group showed significantly lower counts of CD4+T lymphocytes(727.43 ± 138.59/μl vs. 804.25 ± 92.83/μl, t=2.17, P<0.05)and CD4/CD8 ratio(1.23±0.35 vs. 1.46 ± 0.34, t = 2.24, P < 0.05) than the control group, while no significant difference was observed in CD8 + T lymphocyte counts between the CA group and control group(613.60 ± 121.60/μl vs. 572.45 ± 103.08/μl, t=1.24, P>0.05). The levels of serum IL?2 and IFN?γwere both lower in the CA group than in the control group(t=2.12, 2.16, respectively, both P < 0.05). In the CA group, PD?1 and PD?L1 expression levels on peripheral blood CD4 + T lymphocytes were both negatively correlated with CD4+T lymphocyte counts, the CD4/CD8 ratio, as well as IL?2 and IFN?γserum levels(all P<0.05), and those on peripheral blood CD8+T lymphocytes were also negatively correlated with the CD4/CD8 ratio(all P<0.05), but uncorrelated with CD8+T lymphocyte counts(both P>0.05). Conclusion PD?1 was highly expressed on peripheral blood T lymphocytes from patients with CA, which may inhibit T lymphocyte?mediated immune response, decrease CD4+T lymphocyte counts, the CD4/CD8 ratio as well as IL?2 and IFN?γserum levels by interacting with its ligand PD?L1 and forming the PD?1/PD?L1 signaling pathway.

Objective:To investigate the wear characteristics of bovine enamel and lithium disilicate glass ceramic under simulated oral environment.Methods:18 cylindrical lithium disilicate glass-ceramic specimens with the length of 8 mm and diameter of 3 mm were randomly divided into 2 groups (n =9),9 lithium disilicate glass-ceramic specimens and 9 bovine enamel specimens were served as the antagonists respectively.The specimens were then loaded in a wear simulator and subjected to friction force of 10 N for 540 000 cycles in artificial saliva and room temperature(speed 100 r/min,turning radius of 2.5 mm,uniform circular motion) condition.During the testing,10 checkpoints were applied to measure the height loss of the specimens with 3D profilometer,then wear curves were plotted.Scanning electron microscopy were applied to investigate the worn surfaces at different wear stages.Results:At every checkpoints,bovine enamel wear height loss was larger than the lithium disilicate specimens (P ＜ 0.05);bovine enamel wear curve exhibits a runningin period,steady wear period and severe wear period 3 stages of wear,while wear curves of lithium disilicate glass ceramics exhibit onlyrunning-in period and steady wear period 2 wear stages.Both groups had the corresponding micro-morphological features in different periods.Conclusion:Bovine enamel and lithium disilicate glass ceramics show a phase dynamic evolution law under the simulated oral environment.Bovine enamel is more susceptible to wear than lithium disilicate,suggesting that clinical attention should be paid to prevent the excessive wear of natural teeth caused by lithium disilicate glass ceramic restorations.

The expressions of p-STAT3 and osteopontin in 22 cases of normal nevi and 43 cases of malignant melanoma were immunohistochemically detected,and the correlation between p-STAT3 and osteopontin in malignant melanoma and the correlations of p-STAT3 (or osteopontin) with invasion,metastasis and thickness of malignant melanoma were examined.The results showed p-STAT3 was expressed in 2 of 22 cases of normal nevi and 30 of 43 cases of malignant melanoma,while osteopontin was expressed in 3 cases of normal nevi and 29 cases of malignant melanoma.The expressions of p-STAT3 and osteopontin in melanoma were significantly higher than that in benign nevi.There existed significant correlations between the expression of p-STAT3 and that of osteopontin in melanoma.Furthermore,the expression rates of p-STAT3 were significantly higher in invasive or metastatic melanomas than that their non-invasive or non-metastatic counterparts,and the expression rates of osteopontin were significantly higher in invasive melanomas than that in non-invasive ones.It is concluded that p-STAT3 and osteopontin may play important roles in the pathogenesis of malignant melanoma.

In order to investigate the susceptibility of mixed infection of Ureaplasma Urealyticum (UU) and Mycoplasma Hominis (MH) to 7 kinds of antimicrobial agents and comparison with that of UU infection in NGU patients, the in vitro susceptibility was determined by using microdilution method. The positive results were analyzed. The results showed that the sequence of susceptibility to 7 kinds of antimicrobial agents for both UU infection group and UU-MH mixed infection group was almost the same from the highest susceptibility to the lowest accordingly: Josamycin, Doxycycline, Minocycline, Sparfloxacin, Roxithromycin, Ofloxacin and Azithromycin. The total drug resistance rate for UU-MH mixed infection group (97.67%) was significantly higher than that for UU infection group (44.67%, P < 0.01). The highest drug resistance rate in UU group and UU-MH mixed infection group was 31.33% (Ofloxacin) and 90.48% (Azithromycin) respectively. UU-MH mixed infection showed an increased drug resistance and changes of drug resistance spectrum.
Adult ; Anti-Bacterial Agents ; pharmacology ; Azithromycin ; pharmacology ; Doxycycline ; pharmacology ; Drug Resistance, Bacterial ; Female ; Humans ; Josamycin ; pharmacology ; Male ; Middle Aged ; Minocycline ; pharmacology ; Mycoplasma Infections ; complications ; microbiology ; Mycoplasma hominis ; drug effects ; isolation & purification ; Ofloxacin ; pharmacology ; Superinfection ; Ureaplasma Infections ; complications ; microbiology ; Ureaplasma urealyticum ; drug effects ; isolation & purification

In order to investigate the expression of endothelin receptor B (ETR-B) in human malignant melanoma (MM) cells A375 and SK-mel-1 and the proliferative effects of endothelin 3 (ET3) on A375 cells, RT-PCR was applied to detect the expression of ETR-B gene in human MM cells A375 and SK-mel-1. MTT method was used to evaluate the growth enhancing effects of ET3 on A375 cell line in vitro. The results showed that ETR-B gene was expressed in both MM A375 and SK-mel-1 cells. ET3 had stronger ability to enhance the proliferation of A375 cells in vitro in a concentration-dependent manner. It was suggested that ET3/ETR-B might play an important proliferative role in MM.
Cell Line, Tumor ; Cell Proliferation/*drug effects ; Endothelin-3/*pharmacology ; Melanoma/*metabolism ; Melanoma/*pathology ; Receptor, Endothelin B/*metabolism ; Reverse Transcriptase Polymerase Chain Reaction

A complete hospital security system is essential for the life and property safety of medical workers, patients and their families alike. The successful operation of the system depends on a reasonable staffing of the security department.From 2019, a tertiary hospital has explored and practiced the staffing management of security guards based on the actual needs of the hospital. The first job was to classify its security posts setup, and determine the staffing requirements of front-line security posts based on such indicators as scale, risk and people flow. The management range theory was called into play, to set the staffing coefficient of front-line security posts and their corresponding frontline administrators as 7∶1, and set that each management post needs one middle manager. The next job was to calculate the number of security guards per workload/post. The calculations estimated that the number of guards required for the four front-line security posts, namely, gate keeper post, public area patrol post, security and fire control post, and emergency response unit post, was 37, 46, 26 and 26 respectively. The corresponding management posts, namely, the office of the security department, the security management team, the fire management team, and the order maintenance team, required 7, 8, 5 and 5 management personnel respectively, totaling 160. Based on the calculations, the hospital optimized its staffing, and increased its total security personnel from 150 to 160, including the number of gate keeper posts were reduced by 17, and the number of emergency response unit posts was increased by 22. This optimization has effectively empowered the security department in dealing with medical disputes and emergencies, as well as identifying fire hazards, which serves a reference for the rational staffing of hospital security human resources.