Objective To study the development of conus medullaris by quantitative three-dimensional ultrasound in the second and third trimesters of pregnancy,and to analyze the value of terminal caudal vertebral ossification center count in diagnosis of closed spina bifida.Methods A total of 108 normal fetuses in the second and third trimesters of pregnancy who were admitted to Baoding No.1 Central Hospital from January 2019 to March 2024 were selected and assigned to the normal group.The distance between the fetal conus medullaris and the first sacral vertebra,the distance between the end of the conus medullaris and the end of sacrum,and the number of corresponding vertebrae were measured by three-dimensional ultrasound at different gestational weeks.Meanwhile,54 fetuses who were confirmed to have closed spina bifida after delivery were enrolled into the abnormal group.The number of ossification centers on the median sagittal section of the caudal spine was counted and compared between the two groups.The receiver operating characteristic(ROC)curve was drawn to analyze the value of ossification center count of distal caudal vertebrae in diagnosis of closed spina bifida.Results With the increase of gestational weeks,the distance between the fetal conus medullaris and the first sacral vertebra and the distance between the end of the conus medullaris and the end of the sacral bone were gradually increased.The conus medullaris of normal fetuses was mainly located in L1.5,L2 and L2.5,accounting for 85.19%(92/108).The ossification center count of distal caudal vertebra was slightly increased with the increase of gestational weeks in the normal group.At 17-20 gestational weeks,85.71%(12/14)of the ossification center count of distal caudal vertebra was 5 to 7.At 21-28 gestational weeks,100.00%(45/45)of the ossification center count was 6 and more.At week 29 and above,100.00%(45/45)of the ossification center count was more than 7.In the abnormal group,96.30%(52/54)of the fetuses had the ossification center less than 6,and 85.71%(42/49)of the ossification center count was≤5 at 21-32 gestational weeks.There were significant differences in the ossification center count between the two groups at 17-20,21-24,25-28,29-32,and 33-36 gestational weeks(P<0.05).ROC curve showed that,at 17-20,21-24,25-28,29-32,and 33-36 gestational weeks,the area under the curve(AUC)in prediction of closed spina bifida was 0.804,0.744,0.776,0.819,and 0.722 when the ossification center count of distal caudal vertebra was 5.31,4.59,4.81,4.67,and 5.49 as the cut-off value,respectively.Conclusion The conus medullaris moves upward with the increase of gestational weeks in the second and third trimesters of pregnancy.The ossification center count of the distal caudal vertebra is less than 6 in most fetuses with closed spina bifida,which is conducive to screening for closed spina bifida.

By analysing the similarity between defense qi and leukocytes in terms of function， site of action， and circadian rhythm， it is proposed that in traditional Chinese medicine （TCM）， the pathogenesis of leukopenia is defense qi deficiency. By analyzing the relevant discussions on the generation and transmission of defense qi in TCM classics， it is believed that the original qi in lower jiao （焦） is the source of defense qi， while the water and grain qi in middle jiao enriches defense qi， and the upper jiao transmits and distributes defense qi to the whole body. Therefore， when treating leukopenia after chemotherapy with TCM， Guilu Erxian Gelatin （龟鹿二仙胶） and Yougui Pill （右归丸） are often used to tonify the kidney and supplement essence， and moxibustion at Guanyuan （CV 4） and Qihai （CV 6） is usually accompanied to replenish the original qi in lower jiao and enrich the source of defense qi. Guipi Decoction （归脾汤）， Buzhong Yiqi Decoction （补中益气汤）， Shenling Baizhu Powder （参苓白术散）， and Sijunzi Decoction （四君子汤） are often suggested to strengthen spleen and replenish qi， in combination with moxibustion at Zhongwan （CV 12） and Zusanli （ST 36） to transport the spleen and stomach in the middle jiao to enrich the defense qi. Modified Guizhi Decoction （桂枝汤） to harmonize nutrient and defensive aspects is often used， and moxibustion at Dazhui （GV 14） and Feishu （BL 13） or scraping is added to dredge the striae and interstice in the upper jiao and promote transmission and dissemination of the defense qi. Considering the whole process of generation and distribution of defense qi， it is suggested to choose the most appropriate treatment modality flexibly， and combine internal treatment with external treatment， in order to provide ideas for the treatment of leukopenia in tumour patients.

AIM:To establish a nomogram model to predict the effect of serum ferritin on diabetic retinopathy and evaluate the model.METHODS:A total of 21 variables, including ferritin, were screened by univariate and multivariate regression analysis to determine the risk factors of diabetic retinopathy. A nomogram prediction model was established for evaluation and calibration.RESULTS:Ferritin, duration of diabetes, hemoglobin, urine microalbumin, regularity of medication and body mass index were included in the nomogram model. The consistency index of the prediction model with serum ferritin was 0.813(95%CI: 0.748-0.879). The calibration curves of internal and external verification showed good performance, and the probability of the threshold suggested by the decision curve was in the range 10% to 90%. The model had a high net profit value.CONCLUSIONS:Serum ferritin is an important risk factor for diabetic retinopathy. A new nomogram model, which includes body mass index, duration of diabetes, ferritin, hemoglobin, urine microalbumin and regularity of medication, has a high predictive accuracy and could provide early prediction for clinicians.

Objective To explore the dilution conditions and standards in detecting the semen samples with high sperm concentration using computer-aided sperm analysis(CASA)systems.Methods CASA systems with 10 μm depth disposable counting chambers were used for the examination.The samples were divided into undiluted group(Group 1∶sperm concentration＜50×106/mL)and diluted groups(Group 2∶50×106/mL≤ sperm concentration＜100× 106/mL;Group 3∶sperm concentration≥100× 106/mL).When sperm concentration＜50×106/mL,no dilution was performed.When sperm concentration≥50× 106/mL,the samples were diluted with saline at 1∶n/(50×106)ratio(n=sperm concentration,n/[50×106]rounded down)to＜50×106/mL of sperm concentration.The sperm concentration,progressive motility(PR),non-progressive motility(NP),total motility(PR+NP)and immotile sperm percent-age(IM)were analyzed before and after dilution.The consistency of results pre-and post-dilution was compared.ROC curve was used to analyze the optimal dilution threshold.Results The differences in the parameters pre-and post-dilution gradually rosed with the increased sperm concentration.ROC curve analysis showed optimal dilution thresholds were 133.05 × 106/mL,101.75 × 106/mL,118.60×106/mL,90.90×106/mL,111.83×106/mL for the sperm concentration,PR+NP,PR,NP and IM respectively.Considering sperm concentration and NP were most affected the undiluted high concentration samples,the optimal comprehensive dilution threshold was determined as 125.08× 106/mL.Conclusion When sperm concentration exceeds 125×106/mL,it is recommended to dilute semen sample with normal saline.

Objective To investigate the current status of semen testing in the hospitals without assisted reproductive technology in Hu-nan Province,propose improvement strategies to enhance the diagnosis of male infertility and promote reproductive health services in these hospitals.Methods Questionnaire surveys and on-site investigations were conducted to examine the semen testing status in 67 hospitals without assisted reproductive technology in Hunan Province.The existing problems were summarized,and improvement strate-gies were suggested.Results Among the 67 hospitals,59.7％(40/67)performed semen testing.Of these,45％(18/40)possessed dedicated personnel,60％(24/40)possessed staff received relevant training,and 22.5％(9/40)held certificates.Only 30％(12/40)conducted sperm morphology testing,among them 20％(8/40)able to stain and interpret following WHO standards.The hospitals of 37.5％(15/40)used computer-assisted sperm analysis(CASA)systems,and 22.5％(9/40)were equipped with phase-contrast microscopes.30％(12/40)had dedicated examination areas,and 32.5％(13/40)had independent ejaculation rooms,17.5％(7/40)used disposable slides to observe sperm concentration.In 2022,the hospitals of 57.5％(23/40)had an average daily sample volume＜1.The hospital of 67.5％(27/40)performed standard operating procedures,40％(16/40)followed the WHO 5th la-boratory manual for routine testing,15％(6/40)conducted internal quality control,and 12.5％(5/40)participated in external quali-ty assessment(EQA).Another 55％(22/40)wished to participate in EQA.Conclusion The semen testing capacity in the hospitals,in which assisted reproductive technology is not yet carried out currently,urgently requiring multifaceted improvements.The proposed strategies include emphasizing semen testing,establishing reproductive medicine consortia for comprehensive support,establishing a provincial quality control center for EQA,founding a professional committee for a learning platform,offering training to enhance staff expertise,and including the pre-pregnancy semen testing in public health programs.

Objective:To investigate the clinical features and characteristics of gene mutation of patients with Hereditary spherocytosis(HS) and report novel mutations found in this work.Method:We reviewed the literatures for the clinical and genetic features of the 35 patients (24 boys and 11 girls, median age 7.8 years) with HS admitted to Wuhan Children′s Hospital from June 2017 to January 2023. Analyze clinical manifestations the blood routine, and biochemical laboratory results, and use Next-generation sequencing (NGS) to detect pathogenic mutation. Mann-Whitney U testing method was used to analyze the hematological parameters between different genotypes. Results:All of the patients showed clinical manifestations of anemia; 33 children had jaundice, 28 children had splenomegaly, and 11 children had gallstones. Gene mutations were detected in all 35 patients. 17 patients carried ANK1 variants, 14 patients harbored SPTB variants, 1 patient carried SPTA1variants, 1 patient carried SLC4A1 variants, and 2 patients harbored both ANK1 and SPTB variants. A total of 39 heterozygous mutations were found, among which 27 were pathogenic, 6 was likely pathogenic and 6 were of unknown significance according to the ACMG guideline; Among 39 mutations identified, 31 were novel. There were no significant differences in peripheral blood cell parameters and hemolysis indicators between the ANK1 mutant group and the SPTB group (all P>0.05). Conclusion:SPTB and ANK1 mutations are the most common mutations in HS pathogenic genes among patients in Hubei, China, and there was no significant difference in phenotype between ANK1 and SPTB genotype. We found 31 unreported novel variants and expanded the spectrum of variation in this type of disease .

Objective To explore the clinical efficacy of prone position ventilation(PPV)in improving hypoxemia in patients with severe brain damage.Methods A retrospective research method was conducted,140 patients with severe brain damage who were admitted to the department of critical care medicine of the First Affiliated Hospital,Zhejiang University School of Medicine from August 2020 to August 2021 were selected as subject objected.According to the inclusion and exclusion criteria,20 patients with oxygenation index≤200 mmHg(1 mmHg≈0.133 kPa)who were treated with PPV were statistically analyzed.The patients'blood gas analysis related indicators[including arterial partial pressure of oxygen(PaO2),fractional of inspired oxygen(FiO2),oxygenation index,arterial oxygen saturation(SaO2),arterial partial pressure of carbon dioxide(PaCO2),pH value],ventilator-related parameters[including peak inspiratory pressure(PIP),positive end-expiratory pressure(PEEP),tidal volume(VT),lung dynamic compliance(Cdyn),etc.],and mean arterial pressure(MAP),heart rate(HR)were compared before PPV,12 hours after PPV,and 12 hours after reverting to supine position.At the same time,the related complications of patients during PPV were recorded.Results There were 15 males and 5 females,the mean age of the patients was(46.10±17.22)years old,the average PPV time was(22.20±5.94)hours.Compared with before PPV,patients showed significant increases in PaO2,oxygenation index,SaO2,VT,and Cdyn at 12 hours after PPV and 12 hours after recovery from supine position[PaO2(mmHg):98.35±21.85,98.45±17.90 vs.72.15±10.14,oxygenation index(mmHg):198.82±40.51,202.27±46.39 vs.133.20±33.95,SaO2:0.97±0.02,0.97±0.01 vs.0.94±0.03,VT(mL):558.42±111.23,580.29±119.44 vs.484.82±123.77,Cdyn(mL/cmH2O):26.11±5.42,27.90±5.80 vs.24.15±6.13,all P<0.05];Compared with 12 hours after PPV,the Cdyn of the patient still showed a significant increase after 12 hours of recovery from supine position(P<0.05).There were no statistical differences in the FiO2,PaCO2,pH value,PIP,PEEP,HR,and MAP of patients at various time points before and after PPV(all P>0.05).Five patients developed redness and swelling at the skin compression site mainly on the face after PPV,which gradually improved after returning to a supine position.During this period,there was no occurrence of catheter detachment,malignant arrhythmia,or significant hemodynamic instability.Conclusion PPV has a certain clinical effect on improving hypoxemia in patients with severe brain damage.

Objective To investigate the clinical application value of pediatric electronic bronchoscopy on the diagnosis of respiratory disease in the pediatric intensive care unit(PICU)by morphological diagnosis.Methods The clinical data of 124 patients with PICU respiratory critically ill patients who underwent bedside electronic bronchoscopy from July 2017 to December 2022 in Lanzhou University Second Hospital were retrospectively analyzed.Pathogen examination of 96 cases of bronchoalveolar lavage fluid(BALF),bronchoscopy guided nasal endotracheal intubation in 3 cases was conducted.The accuracy of diagnosis of lung lesions was compared between electronic bronchoscopy and chest CT,and the morphological changes of lung lesions under electronic bronchoscopy were observed.Results The accuracy of electronic bronchoscopy in the diagnosis of tracheoesophageal fistula,tracheoesophageal rupture and bronchial foreign body is significantly higher than that of chest CT(tracheoesophageal fistula:4 cases vs.0 case,tracheoesophageal rupture:4 cases vs.0 case,bronchial foreign body:4 cases vs.6 cases,all P<0.05),and there was no significant difference in the accuracy of diagnosis of lung contusion and bronchiectasis between electronic bronchoscopy and chest CT.Typical cases confirmed under endoscopic:4 cases(3.23%)had tracheoesophageal fistula.Bronchial rupture 4 cases(3.23%).Endobronchial tuberculosis was diagnosed in 1 case(0.81%)with bronchial neoplasms and white caseous material attached to the mucosa.Positive bacteria were detected in 23 cases(23.96%).Guided nasotracheal intubation was performed in 3 children with jaw or cervical fracture.Endotracheal catheterization under mechanical ventilation was performed in 13 cases.There were no serious complications and death during and after the operation.Conclusion Pediatric bendable bronchoscopy with PICU is safe and reliable.It can improve the diagnosis rate of respiratory diseases in PICU,and has high clinical application value.

Objective:To explore the comparative study of video laryngoscopy combined with bronchial blocker and video laryngoscopy combined with double-lumen tube in the teaching of endotracheal intubation in thoracic surgery in the standardized residency training of anesthesia.Methods:The trainees of the standardized residency training were randomly divided into control group and experimental group for clinical teaching, with 25 ones in each group. The experimental group was treated with visual laryngoscopy combined with bronchial blocker, while the control group was treated with visual laryngoscopy combined with double-lumen tube group. The intubation time, intubation success rate, positioning time, hemodynamic changes, and complication incidence during intubation, as well as student assessment results were recorded. GraphPad Prism 6.0 was used for t test and Chi-square test. Results:The time of endotracheal intubation [(95.3±10.1) vs. (137.5±13.5)] and positioning time [(100.8±11.7) vs. (155.4±15.3)] in the experimental group were both shorter than those of the control group ( P< 0.001), the hemodynamic changes in patients with immediate intubation were smaller ( P<0.001), the success rate of intubation was higher (92% vs. 68%) ( P<0.001), the complication incidence was lower ( P<0.001) and the students' performance was higher ( P<0.001). Conclusion:In the anesthesia teaching of thoracic surgery, bronchial blocker can reduce the time of endotracheal intubation, lower the hemodynamic changes during intubation, cut down the incidence of complications, improve the success rate of endotracheal intubation and enhance the confidence of students.

Objective:To investigate the mechanism of Candida albicans Int1 in regulating septin organization. Methods:A series of full-length and truncated fragments of Int1 were constructed and fused with green fluorescent protein (GFP). The intracellular localization of the fusion proteins was observed under a fluorescence microscope. The region in Int1 that was required for bud neck localization was identified. Full-length and fragments of Int1 were overexpressed in the yeast Saccharomyces cerevisiae and the changes in cell growth, cell morphology and septin organization were investigated to determine the functional region in Int1 that mediated the interaction with septin. Moreover, the co-localization of the region and septin was analyzed. Results:The full-length Int1 consisted of 1 661 amino acid residues. A middle region of 209 amino acid residues, Int1-M4 (739-947 aa), that could be localized at the bud neck during both small and large bud periods was identified. Overexpression of Int1-M4 led to significant growth defects, elongated bud and disorganized septin. In the cells with elongated bud, Int1-M4 and septin with abnormal structures could be co-localized.Conclusions:Int1-M4 (739-947 aa), the middle region of Int1 containing 209 amino acid residues, mediated the bud neck localization and the interaction with septin, playing an important role in regulating septin organization.