Objective Improving the limb management strategy for patients undergoing coronary intervention through radial artery puncture and observe the application effect,and to provide scientific basis for the prevention of limb complications.Methods From March 2023 to February 2024,patients who underwent coronary intervention in the Cardiovascular Department of a Tertiary A hospital in Wenzhou City were selected.They were randomly di-vided into an experimental group of 241 cases and a control group of 236 cases using a random number table method.After surgery,they were all treated with a rotary hemostatic device for compression hemostasis.The experi-mental group implement improvement strategies for surgical limb management,specifically underwent a reverse Bar-beau test upon returning to the ward after surgery to achieve non-occlusive compression,followed by decompression every hour.The compression intensity of the control group was guided by palpating the distal radial artery pulsa-tion,and the first decompression was performed 1 hour after surgery,followed by decompression at intervals of every 2 hours.The main evaluation indicators are the incidence of surgical limb complications and simplified Chinese version of General Comfort Questionnaire(GCQ)scores in 2 groups of patients,while the secondary evaluation indi-cators are the duration of compression and the number of decompression times in both groups.Results The inci-dence of postoperative complications in the experimental group was 14.11%,which was statistically significant com-pared to 44.49%in the control group(χ2=53.308,P<0.001).The GCQ score of the experimental group was(77.71±5.43)points,which was higher than(74.66±5.83)points in the control group,and the difference was statistically signif-icant(t=-3.354,P=0.001).The compression duration of(172±52)minutes and decompression frequency of 2(2,3)in the experimental group were lower than(289±60)minutes and 4(3,4)in the control group,and the differences were statistically significant(P<0.001).Conclusion The non-occlusive compression method of the radial artery based on the reverse Barbeau test can significantly reduce limb complications in patients undergoing coronary inter-vention,shorten the duration of hemostatic compression,reduce the number of depressions,and improve patient com-fort.It provides objective basis for nursing staff to evaluate the intensity of hemostatic compression and the timing of decompression.

Objective: To improve the social skills of children with ASD by using Program for the Education and Enrichment of Relational Skills(PEERS ), and to reduce the uncertainty towards ASD and negative emotions for mothers of ASD children. Methods: From September to October 2017, 30 dyads of autistic mother and child were recruited and divided into intervention group and control group (15 mother child dyads each). Based on the content of PEERS social skill, cognitive behavior therapy was delivered in group format, through demonstration, role play and group exercise. At the same time, mother child dyads were trained using parallel social technology. Mothers and children with ASD were investigated using Parents Perception of Uncertainty Scale (PPUS), Patient Health Questionnaire 9 (PHQ-9), Chinese Version of the Beck Depression Inventory II(BDI-Ⅱ-C), Beck Anxiety Inventory (BAI), State Trait Anxiety Inventory(STAI-Form Y), and Autism Behavior Checklist (ABC), Cildhood Autism Rating Scale (CARS), and Social Communication Questionnaire (SCQ). Results: Changes in ASD symptom score in children and emotional score of mothers in the intervention group were less than 0. The total score of mother disease uncertainty(74.93±13.58, 90.40± 9.21 ), ambiguity(31.13±7.07, 38.93±4.73), lack of clarity information(11.93±2.09, 13.80±2.54), unpredictability(9.60±1.99, 12.07±2.89), significantly changed after intervention( t =-3.65, -3.55, -2.20, -2.72, P <0.05). Conclusion Social PEERS group intervention can enhance the social skills of children with ASD, reduce uncertainty of illness among mother of ASD children. Timely disease related information, guidance for mothers to actively participate in child care and training, might help to reduce cognitive bias, depressive and anxiety symptoms among mothers.

Background The contracted family doctor services are the embodiment of the implementation of the new medical reform policy, and the transformation of the grass-roots health service mode. Studies have proved that the occupational stress in medical staff was at a high level. The enhancement of professional identity will contribute to strengthen team building，alleviate job burnout, and reduce turnover intention of family doctors. Objective To investigate the current situation of occupational identity among family doctor teams in Chengdu, to examine potential influencing factors of occupational identity, and to provide a reference for promoting career development and team building of family doctor teams. Methods Multi-stage random cluster sampling was adopted to enroll study participants form 46 primary healthcare centers where family doctor contract services were implemented among 23 districts and counties in Chengdu between March 4 and 26, 2021. A total of 2 681 family doctors participated in this survey. A self-reported survey was conducted to collect participants' demographic and occupational data. The Effort-Reward Imbalance (ERI）questionnaire was implemented to assess occupational stress. The Professional Identity Scale was used to appraise occupational identity. Results A total of 2 327 valid questionnaires were collected, with a valid recovery rate of 86.80%, involving 1 715 females (73.7%) and 612 males (26.3%), with dominant age groups of 26−35 years (43.3%) and 36−45 years (30.4%), a high proportion of being married (82.8%), having college (36.0%) and undergraduate (47.3%) education, a high proportion of primary titles (66.0%) and informal work contract (66.1%). About 88.7% of family doctor team workers reported occupational stress. The average score of occupational identity was (3.68±0.62) points. There were significant differences in occupational identity scores among different professional title, work contract, working years in medical institutions, income, and effort/reward ratio (EER) groups (P < 0.05). ERR was negatively correlated with occupational identity (r_s=−0.495, P<0.05）. The multiple regression model showed that occupational identity score in the non-staffed participants was lower than the score in the staffed ones (OR=0.429, 95%CI: 0.299−0.825). The occupational identity score in the participants having associate senior title or above was higher than in without professional title (OR=1.424, 95%CI: 1.194−2.328). The longer the working years, the higher the occupational identity score among the participants. The score of the more than 20 working years group was 1.820 times that of the less than 5 working years group (95%CI: 1.342−2.543). The higher the income, the higher the occupational identity score. The score of the 9001−12000 yuan per month group was 1.977 times that of the 1000−3000 yuan per month group (95%CI: 0.811−9.696) , and the score of the more than 12000 yuan per month group was 2.283 times that of the 1000−3000 yuan per month group (95%CI: 1.199−10.267）. Conclusion The family doctor team workers generally report occupational stress, and their occupational identity is at a medium level in Chengdu. Relevant managers should implement intervention measures against the main influencing factors to reduce their work tension and improve their occupational identity.

Objective:To investigate clinical features, risk factors and prognostic effects of paradoxical response(PR)in children with tuberculous meningitis(TBM)during anti-tuberculosis treatment.Methods:The clinical and follow-up data of TBM children admitted to the Department of Pediatrics, the Affiliated Hospital of Zunyi Medical University between January 2013 and December 2018 were retrospectively analyzed.The children were divided into the PR group and the non-PR group.Influencing factors of PR were selected by the univariate analysis, and independent risk factors were screened from these influencing factors by using the multivariate Logistic regression model.The effect of PR on long-term prognosis (≥9 months) of TBM was evaluated. Results:There were 31 cases(35.6%)with PR among the 87 TBM children enrolled, including 16 boys and 15 girls, with median age of 92(8-168)months.The median time for PR occurrence during the anti-tuberculosis treatment was 33(15-180)days.PR could present dete-rioration or recurrence of original symptoms, cerebrospinal fluid(CSF)deterioration and neuroimaging deterioration, accounting for 71.0%(22/31 cases), 80.6%(25/31 cases)and 51.6%(16/31 cases), respectively.Univariate analysis showed that stage Ⅱ, limb paralysis, cranial nerve palsy, positive tests of tuberculosis infection(T-SPOT), an increased lactate dehydrogenase(LDH)level in CSF, basilar meningeal enhancement, and tuberculosis infection outside the central nervous system were the influencing factors of the PR(all P<0.05). Multivariate analysis showed that limb paralysis, cranial nerve palsy, an increased CSF-LDH level, and positive T-SPOT were independent risk factors of PR(all P<0.05). PR was not associated with prognosis( P=0.165). Conclusions:PR occurs in 35.6% of children with TBM.Limb paralysis, cranial nerve palsy, an increased CSF-LDH level and positive T-SPOT are independent risk factors of PR.PR does not adversely affect the outcome.Identifying PR is extremely important for the prevention of some clinical misunderstandings.

In Gram-negative bacteria, lipopolysaccharide transport (Lpt) protein LptA and LptC form a complex to transport LPS from the inner membrane (IM) to the outer membrane (OM). Blocking the interaction between LptA and LptC will lead to the defect of OM and cell death. Therefore, Lpt protein interaction could be used as a target to screen new drugs for killing Gram-negative bacteria. Here we used biolayer interferometry (BLI) assay to detect the interaction between LptA and LptC, with the aim to develop a method for screening the LptA/LptC interaction blockers in vitro. Firstly, LptC and LptA with or without signal peptide (LptAfull or LptAno signal) were expressed in E. coli BL21(DE3). The purified proteins were then labeled with biotin and the super streptavidin (SSA) biosensor was blocked with diluent. The biotin labeled protein sample was mixed with the sensor, and then the binding of the protein with a series of diluted non biotinylated protein was detected. At the same time, non-biotinylated protein was used as a control. The binding of biotinylated protein to a small molecule IMB-881 and the blocking of interaction were also detected by the same method. In the blank control, the biosensor without biotinylated protein was used to detect the serially diluted samples. The signal response constant was calculated by using steady analysis. The results showed that biotinylated LptC had a good binding activity with LptAfull and LptAno signal with KD value 2.9e⁻⁷±7.9e⁻⁸ and 6.0e⁻⁷±2.8e⁻⁸, respectively; biotinylated LptAno signal had a good binding activity with LptC, with a KD value of 9.6e⁻⁷±7.2e⁻⁸. All binding curves showed obvious fast binding and fast dissociation morphology. The small molecule compound IMB-881 can bind to LptA to block the interaction between LptA and LptC, but has no binding activity with LptC. In summary, we developed a method for detecting the LptA/LptC interaction based on the BLI technology, and confirmed that this method can be used to evaluate the blocking activity of small molecule blockers, providing a new approach for the screening of LptA/LptC interaction blockers.
Carrier Proteins ; Escherichia coli/metabolism* ; Escherichia coli Proteins/metabolism* ; Interferometry ; Membrane Proteins/metabolism*

Targeted programmed death-ligand 1 (PD-L1) and CXC chemokine receptor type 4 (CXCR4), gene sequences encoding anti-PD-L1 nanobody and anti-CXCR4 nanobody were cloned into the pET-22b (+) vector to construct recombinant expression plasmid of anti-PD-L1&CXCR4 bispecific nanobody, which was connected with 6 × His tag and transformed into E.coli BL21 (DE3). The expressed proteins were then found to exist as a soluble form in the supernatant of bacterial lysate after induction of IPTG.Three purification methods were used to obtain the target protein in order to improve the yield and purity of the bispecific nanobody.The bacterial supernatant was separated and purified by His Trap FF affinity chromatographic column.The target protein output could exceed 1 mg/L, and the product purity could reach up to 97%.Besides, the anti-PD-L1&CXCR4 bispecific nanobody shows a specific binding ability to two antigens on the cell surface, enhancing the cytotoxicity of IL-2 activated human peripheral blood mononuclear cells (PBMC) to tumor cell line AsPC-1, which lays the foundation for further evaluation of its drug efficacy in vivo.

Objective To investigate the efficacy and safety of enhanced external counter pulsation(EECP) in treatment of coronary heart disease in patients aged 80 years and older.Methods The 336 consecutive elderly patients with coronary heart disease were treated in our hospital from May 2016 to October 2017 in a prospective study.The 168 coronary heart disease patients aged 80 years and older were divide into a control group(n=84)receiving conventional therapy(anti-platelet drugs,angiotensin converting enzyme inhibitors,angiotensin-receptor blocker,β-blockers,calcium antagonists,nitrates,statins),and EECP group(n =84) receiving enhanced external counter pulsation 0.5 h/time,2 times per day,36 days as a course as add-on therapy to conventional therapy.And 168 patients aged 60～ 79 years received the same therapy as those in EECP group.After one course of EECP treatment,the therapeutic effects were evaluated by Seattle angina questionnaire (SAQ)and Pittsburgh sleep quality index(PSQI),and the incidence of major adverse cardiovascular events and readmission rate were compared between groups of the EECP vs.control group and group of patients aged 60-79 years.Results The scores of SAQ and PSQI were better in the EECP group than in the control group after one course of EECP treatment,as follows:physical limitation(35.7 ±10.7 vs.26.0 ± 12.1),anginal stability (53.3 ± 22.9 vs.33.9 ± 18.9),anginal frequency (39.6 ± 6.5 vs.30.4 ± 21.1),treatment satisfaction(38.0±6.0 vs.33.7±4.7),subjective sleep quality(1.0±0.7 vs.2.5± 0.7),sleep latency(1.3±0.6 vs.2.4±0.5),sleep duration(0.9±0.6 vs.2.3±0.6),habitual sleep efficiency(1.1 ± 0.6 vs.2.5 ± 0.6),sleep disturbances (1.2 ± 0.4 vs.2.2 ± 0.6),use of sleeping medication(1.2±0.6 vs.2.2±0.6),and daytime dysfunction score was 1.3 ±0.5 vs.2.3 ±0.6 (P ＜0.01).The incidence of major adverse cardiovascular events was lower in the EECP group(31.0％,26/84)than in the control group(66.7 ％,56/84)(P＜0.05).At 1,3,6,12 months after EECP treatment,there was no significant difference in the incidence of major adverse cardiovascular events and readmission rate between patients aged 60-79 years vs.80 years and over(P＞ 0.05).Conclusions EECP treatment is relatively safe and effective for coronary heart disease patients aged 80 years and older.

Gram-negative bacteria have become the main pathogens and cause serious clinical problems with increased morbidity and mortality. However, the slow discovery of new antimicrobial agents is unable to meet the need for the treatment of bacterial infections caused by drug-resistant strains. The interaction of L12 and L10 is essential for ribosomal function and protein synthesis. In this study, a yeast two-hybrid system was established to successfully detect the interaction between L12 and L10 proteins from gram-negative bacteria , which allows us to screen compounds that specifically disrupt this interaction. With this system, we identified two compounds IMB-84 and IMB-87 that block L12-L10 interaction and show bactericidal activity against . We used glutathione--transferase (GST) pull-down and surface plasmon resonance (SPR) assays to demonstrate that these compounds disrupt L12-L10 interaction and the target of compounds was further confirmed by the overexpression of target proteins. Moreover, protein synthesis and elongation factor G-dependent GTPase activities are inhibited by two compounds. Therefore, we have identified two antibacterial agents that disrupt L12-L10 interaction by using yeast two-hybrid system.

Objective To investigate the change of serum vitamin D level in the patients with lower respira-tory tract infection and its correlation with lower respiratory tract infection so as to provide reference for clini-cal treatment.Methods 156 cases of lower respiratory tract infection and 60 persons undergoing healthy phys-ical examination in this hospital were collected as the infection group and healthy control group.The sputum specimens from the infection group conducted the bacterial culture and identification.Then the serum vitamin D levels were analyzed in the two groups.Results The serum vitamin D levels in the infection group and healthy control group were 25.76 ng/mL and 49.81 ng/mL,and the difference was statistically significant (P<0.01).In the infection group,serum vitamin D levels in the patients with klebsiella pneumoniae,Acine-tobacter baumannii,Pseudomonas aeruginosa,staphylococcus aureus,Escherichia coli and fungal infection w ere(25.79 ± 7.17),(24.11 ± 10.27),(27.76 ± 9.27),(24.79 ± 6.95),(22.93 ± 3.71)and(30.59 ± 21.31) ng/mL respectively,the difference had statistical significance compared with the control group(P<0.05). Conclusion The level of serum vitamin D in the patients with lower respiratory tract infection is lower than that in healthy population.Its level is associated with lower respiratory tract infection,but the specific mech-anism remains to be further studied.

Objective To study the predictive validity of fidgety general movement assessment in pre-term twins and multiplets for motor development outcomes.Methods A total of 53 pre-term twins or multiplets delivered between July 2011 and February 2016 participated in this study.They were assessed using a general movements (GM) assessment and participated in the follow-up program until one year old.The motor development outcomes of the infants at one year old were determined according to clinical diagnoses and the Peabody developmental motor scale number two (PDMS-2) evaluation.The predictive validity of fidgety general movement assessment for motor development outcomes was calculated against the standard motor development of infants at one year old.Results There were 53 twins or multiplets who accepted the GM assessment of fidgety movement period.Of these,43 were assessed as normal (NF) and ten (19％) as lacking a normal level of fidgety movement (F-).All 53 cases were followed-up for the motor development outcome.Forty-three cases (81.1％) were assessed as normal at one year old,while ten (18.9％) were assessed as abnormal.All ten had cerebral palsy,and no motor development retardation was found.The predictive value of F-for cerebral palsy was 90.0％ in terms of sensitivity,97.7％ in terms of specificity,90.0％ in positive predictive value,and 97.7％ in terms of negative predictive value.Conclusions Among pre-term twins or multiplets,the fidgety general movement assessment can be a useful early indicator of motor development difficulties.