Objective:To establish an in vivo diffusion model of Treponema pallidum (Tp) in New Zealand rabbits. Methods:A standard strain of Tp (Nichols strain) was recovered in the testes of New Zealand rabbits, and isolated and passaged continuously. The suspensions of the second-passage Tp were collected and inoculated onto the dorsal skin of New Zealand rabbits. After 21-day infection, the New Zealand rabbits were anesthetized and sacrificed, blood samples were collected, and skin tissues at the infection site as well as liver, spleen, testes and lymph nodes were aseptically resected. Real-time fluorescence-based quantitative PCR was performed to detect the spread of Tp in different tissues and organs.Results:On day 21 after infection with Tp, skin lesions such as indurations and ulcers were seen at all inoculated sites of New Zealand rabbits. Pathological examination showed a lot of inflammatory cells in the infected lesions, mainly including plasma cells, macrophages and lymphocytes. Real-time fluorescence-based quantitative PCR revealed a large number of Tp in tissues and organs, such as liver, spleen and testes.Conclusion:After inoculation with Tp in the dorsal skin of New Zealand rabbits, Tp could spread to the liver, spleen, testes and other tissues and organs through blood and lymph nodes, and the in vivo diffusion model of Tp strains in New Zealand rabbits was successfully constructed.

OBJECTIVETo obtain sufficient recombinant VP2 protein of human Bocavirus and establish it's seroepidemiology assying metbord. METHORD: Tbe capsid protein VP2 DNA genes of HBoV1 and 2 were optimized in accordance with tbe usage of the favorite codons in K coil so as to enhance its protein expression in prokaryotic expressing system. The protein was purified by Ni-NTA column, and its antigenicity was determined by Western Blot. Then establish ELISA to detect the specific anti-VP2 IgG antibodies against HBoV1 and 2 in healthy children aged 3-6 years in Nanjing, China.

RESULTSThe recombinant protein 6 x His-VP2 was produced in a larger quantity at 25 degrees C induced by IPTG (1 mmol/L) over night and purified by Ni-NTA column. Seropositive rates of HBoV1 and 2 were 62.2% and 55.5% and their mixed seropositivity was 37%.

CONCLUSIONThe optimizing expression of the capsid protein VP2 from human Bocavirus constructed successfully and get a high yield under certain conditions. The established ELISA could be used to further analyze seroepidemiology of HBoV in China.

Capsid Proteins ; analysis ; genetics ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Human bocavirus ; isolation & purification ; Humans ; Male ; Polymerase Chain Reaction ; Seroepidemiologic Studies

BACKGROUND: Posterior pedicle screw fixation of lower cervical spine has many advantages. However, because of the special anatomy of pedicle, the technical requirements of nailing placement are high and it is also a challenge to surgeons. The application of three-dimensional (3D) printing technology can help to complete screw placement, and has certain advantages. OBJECTIVE: To introduce the method of 3D printing personalized navigation template assisted posterior lower cervical pedicle screw fixation and evaluate its clinical application value. METHODS: Totally 20 patients with lower cervical spine pedicle screw internal fixation who were treated from June 2015 to December 2016 in Affiliated Hospital of Yan'an University were retrospectively analyzed. Before the surgery, the patient's lower cervical spine CT data was introduced into the relevant software and the 3D printing personalized navigation template assisted nailing pedicle screw was designed. The number of screws inserted was recorded. After the operation, CT scan was used to calculate the accuracy rate of screw placement. Preoperative and postoperative cervical spine Visual Analogue Scale score and spinal function score of Japanese Orthopedic Association were used to evaluate the clinical effect of cervical nerve. RESULTS AND CONCLUSION: (1) All patients were followed up for 3 months postoperatively. (2) 3D printing personalized navigation template assisted 132 pedicle screws. The precise insertion rate was 94.7%. Among them, 125 screws were at grade 0 (94.7%, 125/132), 3 screws grade 1 (2.3%, 3/132), 4 screws grade 2 (3.0%, 4/132), no screw grade 3. (3) At postoperative 1 and 3 months, the neck shoulder Visual Analogue Scale score and spinal function score of Japanese Orthopedic Association of the patients were significantly improved (P < 0.05), but there was no significant difference (P > 0.05), compared with those at preoperative. (4) In summary, 3D printing personalized navigation template assisted cervical pedicle screw placement had the high insertion rate and satisfactory clinical efficacy.

Objective:To establish an ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for plasma caffeine concentration detection, and to explore the clinical value of caffeine therapeutic drug monitoring (TDM) in the treatment of premature infants with respiratory distress syndrome (RDS).Method:Take the plasma sample in a centrifuge tube, add the caffeine deuterated isotope internal standard, then add the protein precipitant, vortex the mixture thoroughly, and centrifuge the supernatant to enter the mass spectrometry analysis. The mobile phase were methanol and water, gradient elution; the column temperature was 45 ℃, the method was established using Shimadzu LC-30AD-CL liquid system and AB SCIEX 4500 QTRAP mass spectrometer, and the sensitivity, specificity, linearity, accuracy imprecision, matrix effect, and carry-over of the method were evaluated. Sample from 30 patients diagnosed with neonatal RSD were collected in the Department of Neonatology of Renmin Hospital of Wuhan University from February to April 2021, then detected the trough concentration of caffeine in premature infants with RDS after taking the same dose of caffeine to assess the impact of individual variation on caffeine drug concentration.Results:The detection limit of caffeine was 0.02 μg/ml, and the lowest limit of quantification was 0.05 μg/ml. It showed good linearity ( R2=0.9986, R>0.99) in the concentration range from 1.0 to 100.0 μg/ml, specificity (recovery rate of 85.52%-114.12%), accuracy (recovery rate 85.97%-114.53%), intra-day and inter-day imprecision ( CV 6.01%-11.28%), matrix effects and carryover pollution were negligible. The trough concentration of 30 preterm infants with RSD after taking the same dose of caffeine (10 mg/kg) was (25.45±11.61) μg/ml, and the coefficient of variation was 44.88%. Conclusion:This study established an accurate and reliable UPLC-MS/MS method with low sample consumption to monitor the blood concentration of caffeine; caffeine TDM has certain clinical application value, which can be used to assist RDS diagnosis and treatment and improve the efficacy of caffeine.

Objective:To explore the correlation between time in range (TIR) after short-term treatment and glycated hemoglobin after 3 months (HbA lc-3m) in patients with newly-diagnosed type 2 diabetes mellitus (T2DM). Methods:In this cross-sectional study, a total of 94 patients with newly-diagnosed T2DM who received treatment in the Department of Endocrinology of Inner Mongolia Autonomous Region People′s Hospital were enrolled from January 2018 to September 2022. The patients were followed-up for 3 months and had complete medical record. TIR was divided into three groups according to different target ranges of blood glucose (TIR1: TIR with blood glucose between 3.9 and 10.0 mmol/L, TIR2: TIR with blood glucose between 3.9 and 7.8 mmol/L, TIR3: TIR with fasting, premeal or bedtime blood glucose <6.1 mmol/L and 2 h postprandial blood glucose <8.0 mmol/L). The patients were divided into two groups based on whether their HbA 1c-3m level was less than 6.5%, and the baseline data and variations in TIR for distinct target glucose levels were compared between the two groups. Spearman′s correlation analysis and binary logistic regression analysis were used to analyze the relationship between baseline indicators, TIR after short-term treatment and HbA 1c-3m. Receiver operating characteristic curve (ROC) was drawn to evaluate the predictive ability of different TIR after short-term therapy for HbA 1c-3m. Results:There were statistically significant differences in TIR1 [81.0 (67.5, 94.6)% vs 71.4 (51.7, 85.7)%], TIR2 [57.7 (29.7, 70.8)% vs 40.9 (22.4, 52.3)%] and TIR3 [23.8 (10.2, 39.5)% vs 13.0 (4.8, 25.0)%] between patients with a HbA 1c-3m<6.5% and patients with a HbA 1c-3m≥6.5% (all P<0.05). Spearman correlation analysis showed that among all the patients with newly-diagnosed T2DM, TIR1, TIR2 and TIR3 were all negatively correlated with HbA 1c-3m [6.4 (6.1, 6.9)%] ( r=-0.322, -0.348, -0.303, respectively, all P<0.01). Logistic regression analysis showed that after adjusting for the confounding factors, TIR1 ( OR=1.021, 95% CI: 1.002-1.041; P=0.034), TIR2 ( OR=1.024, 95% CI: 1.006-1.043; P=0.011), TIR3 ( OR=1.037, 95% CI: 1.010-1.065; P=0.008) were all independently related to HbA 1c-3m. When HbA lc-3m<6.5% was taken as the target value, the area under the ROC curve: TIR1 was 0.639 (95% CI: 0.528-0.751), TIR2 was 0.671 (95% CI: 0.560-0.782), TIR3 was 0.659 (95% CI: 0.549-0.770), respectively. When HbA lc-3m<7.0% was taken as the target value, the area under the ROC curve: TIR1 was 0. 730 (95% CI: 0.619-0.841), TIR2 was 0.744 (95% CI: 0.642-0.846), TIR3 was 0.701 (95% CI: 0.588-0.814). There was no significant difference in the area among the three statistics ( P>0.05). Conclusions:For newly-diagnosed T2DM patients, TIR after short-term treatment is negatively correlated with HbA 1c after 3 months and has good predictive value for it.

OBJECTIVE: To explore the genotyping characteristics of human fecal Escherichia coli( E. coli) and the relationships between antibiotic resistance genes (ARGs) and multidrug resistance (MDR) of E. coli in Miyun District, Beijing, an area with high incidence of infectious diarrheal cases but no related data. METHODS: Over a period of 3 years, 94 E. coli strains were isolated from fecal samples collected from Miyun District Hospital, a surveillance hospital of the National Pathogen Identification Network. The antibiotic susceptibility of the isolates was determined by the broth microdilution method. ARGs, multilocus sequence typing (MLST), and polymorphism trees were analyzed using whole-genome sequencing data (WGS). RESULTS: This study revealed that 68.09% of the isolates had MDR, prevalent and distributed in different clades, with a relatively high rate and low pathogenicity. There was no difference in MDR between the diarrheal (49/70) and healthy groups (15/24). CONCLUSION We developed a random forest (RF) prediction model of TEM.1 + baeR + mphA + mphB + QnrS1 + AAC.3-IId to identify MDR status, highlighting its potential for early resistance identification. The causes of MDR are likely mobile units transmitting the ARGs. In the future, we will continue to strengthen the monitoring of ARGs and MDR, and increase the number of strains to further verify the accuracy of the MDR markers.
Humans ; Escherichia coli/genetics* ; Escherichia coli Infections/epidemiology* ; Multilocus Sequence Typing ; Genotype ; Beijing ; Drug Resistance, Multiple, Bacterial/genetics* ; Anti-Bacterial Agents/pharmacology* ; Diarrhea ; Microbial Sensitivity Tests

OBJECTIVES: To investigate the clinical treatment outcomes and the changes of the outcomes over time in extremely preterm twins in Guangdong Province, China. METHODS: A retrospective analysis was performed for 269 pairs of extremely preterm twins with a gestational age of <28 weeks who were admitted to the department of neonatology in 26 grade A tertiary hospitals in Guangdong Province from January 2008 to December 2017. According to the admission time, they were divided into two groups: 2008-2012 and 2013-2017. Besides, each pair of twins was divided into the heavier infant and the lighter infant subgroups according to birth weight. The perinatal data of mothers and hospitalization data of neonates were collected. The survival rate of twins and the incidence rate of complications were compared between the 2008-2012 and 2013-2017 groups. RESULTS: Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of severe asphyxia and smaller head circumference at birth (P<0.05). The mortality rates of both of the twins, the heavier infant of the twins, and the lighter infant of the twins were lower in the 2013-2017 group compared with the 2008-2012 group (P<0.05). Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of pulmonary hemorrhage, patent ductus arteriosus (PDA), periventricular-intraventricular hemorrhage (P-IVH), and neonatal respiratory distress syndrome (NRDS) and a higher incidence rate of bronchopulmonary dysplasia (P<0.05). CONCLUSIONS There is a significant increase in the survival rate over time in extremely preterm twins with a gestational age of <28 weeks in the 26 grade A tertiary hospitals in Guangdong Province. The incidences of severe asphyxia, pulmonary hemorrhage, PDA, P-IVH, and NRDS decrease in both the heavier and lighter infants of the twins, but the incidence of bronchopulmonary dysplasia increases. With the improvement of diagnosis and treatment, the multidisciplinary collaboration between different fields of fetal medicine including prenatal diagnosis, obstetrics, and neonatology is needed in the future to jointly develop management strategies for twin pregnancy.
Bronchopulmonary Dysplasia/epidemiology* ; Female ; Gestational Age ; Humans ; Infant ; Infant, Extremely Premature ; Infant, Newborn ; Pregnancy ; Respiratory Distress Syndrome, Newborn/epidemiology* ; Retrospective Studies ; Treatment Outcome

Objective: To investigate the effect of casein kinase 2 interacting protein-1 (CKIP-1) on craniofacial soft tissues and hard tissues, to provide the basis for the study and treatment of craniomaxillofacial related diseases. Methods: 6-month- old male CKIP-1 knockout (KO) mice were selected as the experimental group, and wild-type (WT) mice were selected as the control group. The craniomaxillofacial hard tissues (parietal bone, nasal bone, incisors and molars) were analyzed through micro- CT, and the morphological changes of maxillofacial soft tissues (nasal cartilage, lip mucosa and tongue) were analyzed through HE staining and toluidine blue staining. Results: CKIP-1 negatively regulated bone mass of cancellous bone of cranial and maxillofacial bones and dentin mineralization. Compared with the WT mice, the thickness of the parietal baffle layer increased by 93% in KO mice, while cortical bone showed no significant difference between the two groups. The nasal cancellous bone thickness increased by 160% in KO-mice, while cortical bone showed no significant difference between the two groups; the enamel thickness was normal, but the pulp cavity became smaller and the dentin thickness increased by 48%. Compared with the WT mice, the HE staining and toluidine blue staining analyses of the soft tissues revealed that the thickness of the alar cartilage plate of KO mice increased by 57%, and local ossification was found within the cartilage plate. The thickness of the keratinized layer of the labial mucosa increased by 170% in KO mice and the muscle fiber diameter of the lingual muscle increased by 45%. Conclusion CKIP-1 genes have different effects on the growth and development of various soft and hard tissues in the maxillofacial region of mice.