OBJECTIVE To standardize the main processes and related technical links of the clinical comprehensive evaluation of drugs， and provide guidance and reference for improving the quality of comprehensive evaluation evidence and its transformation and application value. METHODS The construction of Guideline for the Workflow of Clinical Comprehensive Evaluation of Drugs was based on the standard guideline formulation method of the World Health Organization （WHO）， strictly followed the latest definition of guidelines by the Institute of Medicine of the National Academy of Sciences of the United States， and conformed to the six major areas of the Guideline Research and Evaluation Tool Ⅱ. Delphi method was adopted to construct the research questions； research evidence was established by applying the research methods of evidence-based medicine. The evidence quality classification system of the Chinese Evidence-Based Medicine Center was adopted for evidence classification and evaluation. The recommendation strength was determined by the recommendation strength classification standard formulated by the Oxford University Evidence-Based Medicine Center， and the recommendation opinions were formed through the expert consensus method. RESULTS & CONCLUSIONS The Guideline for the Workflow of Clinical Comprehensive Evaluation of Drugs covers 4 major categories of research questions， including topic selection， evaluation implementation， evidence evaluation， and application and transformation of results. The formulation of this guideline has standardized the technical links of the entire process of clinical comprehensive evaluation of drugs， which can effectively guide the high-quality and high-efficient development of this work， enhance the standardized output and transformation application value of evaluation evidence， and provide high-quality evidence support for the scientific decision-making of health and the rationalization of clinical medication.

Objective:To investigate the clinical efficacy of labial mucosa graft coupled preputial island flap urethroplasty for recurrent penile curvature after hypospadias repair.Methods:The data of 35 patients with recurrent penile curvature after hypospadias repair who underwent labial mucosa graft coupled preputial island flap urethroplasty in four hospitals from January 2021 to March 2023 were retrospectively reviewed. The median age of the patients was 15(13, 18)years old, five of them had penile dehiscence, and four had urethrocutaneous fistula. All patients received labial mucosa graft coupled preputial island flap urethroplasty, the penile flexion was confirmed by artificial erection test, and the urethra is obliquely cut at the most obvious curvature of the penis. By removing the ventral fibrotic scar tissue, the length of the urethral defect was measured. The urethral plate was reconstructed with the same length of labial mucosa graft and an adjacent preputial island flap was transplanted to cover the reconstructed urethra plate to form a new urethral lumen. The inner urethral stent tube was extracted three days after the operation, and the outer urethral stent tube was removed three weeks after the surgery. Diagnostic urethral exploration was performed 5 weeks post-operation, uroflowmetry examination was performed seven weeks after the procedure, and the perioperative and postoperative follow-up data were analyzed.Results:All patients underwent a successful surgery, with the average duration of the procedure being (114.7±20.1)minutes. After mean follow-up of (15.5±5.9)months, no penile curvature occurred, and 32 patients were successfully repaired. Uroflowmetry examination was achieved in 21 patients at the seventh week after surgery showed their mean maximum flow rate was (18.4±3.7)ml/s. Three cases (8.6%, 3/35) developed urethrocutaneous fistula, who were repaired successfully 6 months after surgery.Conclusions:Utilizing the combination of labial mucosa grafts and preputial island flaps for urethroplasty has been proven to be a reliable and safe option for individuals with recurrent penile curvature following hypospadias repair, with a high success rate and few complications.

Nerve regeneration in adult mammalian spinal cord is poor because of the lack of intrinsic regeneration of neurons and extrinsic factors - the glial scar is triggered by injury and inhibits or promotes regeneration. Recent technological advances in spatial transcriptomics (ST) provide a unique opportunity to decipher most genes systematically throughout scar formation, which remains poorly understood. Here, we first constructed the tissue-wide gene expression patterns of mouse spinal cords over the course of scar formation using ST after spinal cord injury from 32 samples. Locally, we profiled gene expression gradients from the leading edge to the core of the scar areas to further understand the scar microenvironment, such as neurotransmitter disorders, activation of the pro-inflammatory response, neurotoxic saturated lipids, angiogenesis, obstructed axon extension, and extracellular structure re-organization. In addition, we described 21 cell transcriptional states during scar formation and delineated the origins, functional diversity, and possible trajectories of subpopulations of fibroblasts, glia, and immune cells. Specifically, we found some regulators in special cell types, such as Thbs1 and Col1a2 in macrophages, CD36 and Postn in fibroblasts, Plxnb2 and Nxpe3 in microglia, Clu in astrocytes, and CD74 in oligodendrocytes. Furthermore, salvianolic acid B, a blood-brain barrier permeation and CD36 inhibitor, was administered after surgery and found to remedy fibrosis. Subsequently, we described the extent of the scar boundary and profiled the bidirectional ligand-receptor interactions at the neighboring cluster boundary, contributing to maintain scar architecture during gliosis and fibrosis, and found that GPR37L1_PSAP, and GPR37_PSAP were the most significant gene-pairs among microglia, fibroblasts, and astrocytes. Last, we quantified the fraction of scar-resident cells and proposed four possible phases of scar formation: macrophage infiltration, proliferation and differentiation of scar-resident cells, scar emergence, and scar stationary. Together, these profiles delineated the spatial heterogeneity of the scar, confirmed the previous concepts about scar architecture, provided some new clues for scar formation, and served as a valuable resource for the treatment of central nervous system injury.
Mice ; Animals ; Gliosis/pathology* ; Cicatrix/pathology* ; Spinal Cord Injuries ; Astrocytes/metabolism* ; Spinal Cord/pathology* ; Fibrosis ; Mammals ; Receptors, G-Protein-Coupled

【Objective】 To verify the key performance parameters of Roche Cobas S201 multiplex NAT for blood donors, so as to provide data for the conformation of multiplex NAT performance and establish a scientific and feasible verification scheme. 【Methods】 Comprehensive performance verification on four key parameters of ROCHE COBAS S201 multiplex NAT system, including the 95% detection limit, compliance rate, anti-interference ability and operator comparison were conducted. The data were compared with the specifications provided by the manufacturer to evaluate the performance of laboratory testing system and ensure the quality of blood testing. 【Results】 2 sets of COBAS S201 multiplex NAT system in our laboratory were used to perform 5 times of the 95% detection limit value declared by the manufacturer for individual tests. The experimental results showed that the target NAT can be detected 100% and the lower limit of determination was verified. Single-virus NAT was performed to detect 5 different HBV DNA concentrations (25 ~400 IU/mL), 5 different HCV RNA concentrations (25 ~400 IU/mL), 5 different HIV RNA Concentration (100~1 000 IU/mL), and 5 negative tubes. The experimental results showed that all detected values of 20 tubes were 100% consistent with the true value, and the performance parameter "coincidence rate" was verified. Samples containing lipoemia, hemolysis, with ALT>50 mL/L and syphilis-positive endogenous interfering substances with 3 times of the 95% detection limit value were tested for 3 times, and results showed both response rate (3/3) and yielding rate reached 100%. In the control group (with none or in the normal range of interfering substances), reactivity was detected in samples with extremely low concentration of 3 times of the 95% detection limit, showing no significant difference (P>0.05). Above results fully showed that the existence of interference substances (lipidemia, hemolysis, ALT ≥ 50 and syphilis positive) in the blood of donors had no significant effect on the yielding of HBV/HCV/HIV virus target nucleic acids. Different operators performed the sample loading tests with same instrument, reagent and specimen showed 100% consistency in the results. It could be preliminarily assessed that there was no difference in the operations between the operators in this verification. 【Conclusion】 The verification scheme of the multiplex NAT system for methodology performance index (95% detection limit, coincidence rate, anti-interference ability, and operator comparison) established in this study, showed simple, flexible, scientific and feasible characteristics and could provide reference and data support for domestic blood transfusion services in acid detection.

Objective To evaluate the efficacy and safety of chloral hydrate combined with dexmedetomidine for sedation during echocardiography in pediatric patients with Williams-Beuren syndrome.Methods Eighteen pediatric patients diagnosed with Williams-Beuren syndrome by genetic testing,aged 5-58 months,scheduled for elective echocardiography under sedation,received oral chloral hydrate 50 mg/kg.Vital signs were measured every 5 min,and sedation was assessed using Ramsay sedation score.When Ramsay sedation score＜4 points 20 min later,intranasal dexmedetomidine 1 μg/kg was given as rescue sedative.Medicine used,vital signs,onset time,moderate and deep sedation duration and emergence time were recorded.Results The success rate of sedation with chloral hydrate alone was 38.9％ and with chloral hydrate and dexmedetomidine 61.1％.The onset time,sedation duration and emergence time were (15.7+1.9) min,(75+26) min and (52+25) min,respectively,in pediatric patients received chloral hydrate alone.The onset time,sedation duration and emergence time were (33.2±3.4) min,(83±49) min and (61±46) min,respectively,in pediatric patients received chloral hydrate and dexmedetomidine.The onset time was significantly prolonged in pediatric patients received chloral hydrate and dexmedetomidine than in pediatric patients received chloral hydrate alone (P＜0.05).Heart rate,respiratory rate and SpO2 were stable during sedation in all pediatric patients,and nausea and mild vomiting were found in 3 pediatric patients received chloral hydrate and in 6 pediatric patients received chloral hydrate and dexmedetomidine,and no other adverse reactions were observed.Conclusion Oral chloral hydrate 50 mg/kg combined with intranasal dexmedetomidine 1 μg/kg provides reliable sedative efficacy and exerts less influence on respiratory and circulatory function with higher safety when used for echocardiography in pediatric patients with Williams-Beuren syndrome.

Objective To evaluate the accuracy of ultrasonographic measurement of the subglottic airway diameter in selecting the cuffed endotracheal tube (ETT) size for the pediatric patients with congenital heart disease.Methods Sixty pediatric patients of both sexes with congenital heart disease,of American Society of Anesthesiologists physical status Ⅱ-Ⅳ,aged 1 month-7 yr,undergoing elective open heart surgery,were included in this study.The subglottic airway diameter was measured by bedside ultrasonography after induction of anesthesia.The outer diameter of the cuffed ETT was determined according to the subglottic airway diameter.The air leak test was performed after intubation to determine whether or not the ETT size selected based on ultrasonography was appropriate.When the difference between the inner diameter of the finally selected ETT and the result calculated by the Motoyama formula was less than 0.2 mm,the ETT size selected based on the Motoyama formula was considered appropriate.The agreement and correlation between the subglottic airway diameter measured by age-based formula and ultrasonography and the outer diameter of the actually selected ETT were analyzed using Bland-Altman plot and Passing-Bablok regression analysis.Results The accurate rate of the ETT selected based on the ultrasonic measurement and Motoyama formula were 80％ and 55％,respectively,and there was significant difference (P＜0.05).Conclusion Ultrasonographic measurement of the subglottic airway diameter produces higher accuracy than the classical Motoyama formula in selecting the cuffed ETT size for the pediatric patients with congenital heart disease.

Objective:To detect the effect of microRNA-7 ( miR-7 ) knockdown on pathology in murine acute lung injury ( ALI) model,and preliminarily explore its significance.Methods:Murine ALI model was performed by intraperitoneal injection of Li-popolysaccharide (LPS) (10 mg/kg) into miR-7KD mice and wild-type (wild type,WT) mice respectively.Then,the pathologic injury of lung tissue were observed by HE staining.And total cell count of bronchoalveolarlavage(BAL) was calculated.The relative expression of related cytokines in lung tissue was analyzed by Real-time PCR assay.Furthermore,the changes on proportion of innate immune cells (γδT cell and F4/80 macrophages cell) and adaptive immune cell ( CD4+T cell and CD8+T cell) were analyzed by FACS.Meanwhile, the expression of CD62L and CD69,as well as the absolute number,in CD4+T cell were also analyzed.Results: Compared with WT mice,pathological damage in lung tissues was significantly alleviated in miR-7KD mice.Real-time PCR analysis showed that the relative expression of IL-6 was obviously reduced (P<0.01),conversely,relative expression of IL-4 and TGF-βwere obviously increased (P<0.05).Furthermore,the total cell number in BAL also reduced significantly (P<0.05).Importantly,FACS analysis showed that the proportion and the absolute number of F4/80+Mφcells obviously reduced (P<0.05);however,the proportion of γδT cells increased (P<0.05).Moreover,the proportion and the absolute number of CD4+T cells and CD8+T cells were significantly reduced (P<0.05). Finally, the proportion and the absolute number of CD62L+in CD4+T cells were upregulated vigorously,contrastly,the proportion and the absolute number of CD69+in CD4+T cells were notably up-regulated (P<0.05).Conclusion:miR-7 defeciency could significantly ameliorate the pathology of murine ALI,suggesting that it may play an important regulatory role in the development of ALI.

Objective:To detect the change of composition of immune cells in the spleen of miR-126 knockdown(miR-126KD) mice and preliminarily explore its significance .Methods: The expression level of miR-126 in spleens of miR-126KD mice was deter-mined by Realtime PCR.And the total number of splenocytes was calculated.The pathologic morphology change of the spleen was observed by HE staining.And the changes on proportion of DCs ,macrophages cells ,γδ T cells and NK T cells,CD3+T cells and its subgroup ,as well as CD19+B cells in spleens of miR-126KD mice were analyzed by Flow cytometry and calculated respectively.The level of phosphorylated AKt and NF-κB was analyzed by Western blot assay.Results:Compared with those of WT mice ,the expression level of miR-126 decreased obviously ( P<0.05 ) and the total number of cells increased obviously in spleen of miR-126 KD mice ( P<0.05).Moreover,the pathologic morphology of miR-126KD mice was significantly changed.The proportion and number of NK T cells in the inherent cells were significantly increased ( P<0.05 ) , but the proportion of Mφcells were significantly decreased ( P<0.05 ) . Meanwhile,the proportion and number of CD3+T cells and CD4+T cells in the adaptive immune cells were significantly increased (P<0.05),while the total number of CD19+B cells were significantly decreased (P<0.05).Last,the level of phosphorylation Akt and NF-κB increased obviously in spleen of miRNA-126 knockdown mice.Conclusion: Change obviously on the composition of immune cells subsets in the spleen of miRNA-126 knockdown mice ,which it may be related to the altered level of phosphorylated AKt and NF-κB and provides a preliminary experimental basis for the further exploring the roles of miR -126 in immune response.

Objective:To detect the change of proportions of αβT lymphocytes in mesenteric lymph nodes of miR-7 knockdown (miR-7KD)mice,and preliminarily explore its importance.Methods: The volume,weight index and total cells number of mesenteric lymph node in miR-7KD mice were measured.The pathologic morphology change of mesenteric lymph nodes was observed by HE stai -ning.And the changes on proportion of αβT lymphocytes in mesenteric lymph nodes of miR-7KD mice were analyzed by Flow cytometry.Results:Compared with those of WT (wild type) mice,the volume,weight index,and the total cells number of mesenteric lymph node were significantly increased ( P<0.05 ).Moreover ,the pathologic morphology was significantly changed.The proportion and numbers of T lymphocyte were significantly increased;however,the proportion of B lymphocyte were significantly decreased (P<0.05). Notably,the proportion and number of CD4+T cells and CD8+T cells were significantly increased (P<0.05).Meanwhile,CD62L+T cell proportion were vigorously reduced and CD 69+T cell and IFN-γ+T cell proportion were notably up-regulated,which belong to CD4+and CD8+T cell(P<0.05).Conclusion:There was significant influence in the proportion of αβT lymphocytes in mesenteric lymph nodes of miR-7KD mice,suggesting that miR-7 might play an important role in the composition and function of lymphocytes in mesenteric lymph nodes.