Objective:To explore the feasibility of joint screening of the two primary immunodeficiency diseases [severe combined immunodeficiency (SCID) and X-linked agammaglobulinemia(XLA)] and spinal muscular atrophy(SMA) in newborns by multiplex real-time quantitative PCR technology, and to provide evidence for early screening, diagnosis and treatment of children.Methods:Cross-sectional study. From July 2021 to January 2023, a total of 103 240 dry blood spots samples of newborns were collected which were delivered to Neonatal Disease Screening Center of Zhejiang by cold chain transportation. The concentrations of the T cell receptor excision ring (TREC), Kappa deletion of the recombinant excision loop (KREC), and exon 7 deletion of Survival Motor Neuron 1 (SMN1) gene in dry blood spots were simultaneously detected by multiplex real-time fluorescence quantitative PCR, taken ribonuclease P/MRP 30 000 subunits (RPP30) as an internal reference gene. The positive newborns were further diagnosed by other laboratory tests and gene sequencing was taken as gold standard. Children samples from 1 case of SCID, 3 cases of XLA and 2 cases of SMA were used for positive verification. The correlation between detected concentration of TREC/KREC and basic information in newborns were analyzed. The differences among groups for each factor were analyzed.Results:One case of SCID, 2 cases of XLA, 9 cases of SMA and 7 cases of other genetic diseases (4 cases of DiGeorge syndrome, 1 case of trisomy 21 syndrome, 1 case of Noonan syndrome and 1 case of super male syndrome) were identified by multiplex real-time fluorescence quantitative PCR. The positive predictive values of screening neonatal SCID, XLA and SMA were 2.44% (1/41), 2.78% (2/72) and 9/9 respectively. Taking the samples from clinically diagnosed 1 case of SCID, 3 cases of XLA and 2 cases of SMA as positive validation samples, which were all identified. The detected results of TREC/KREC correlated with time of blood collection, sex, weight, gestational age and delivery mode of newborns, whose r values were 0.162/0.187, 0.066/0.032, 0.045/0.042, ?0.015/?0.088 and 0.014/0.068 respectively (all P<0.05). Conclusions:Relying on current neonatal screening platform in Zhejiang, it is feasible to screen jointly two kinds of primary immunodeficiency diseases and spinal muscular atrophy in newborns by multiple real-time fluorescence quantitative PCR technology.

Objective:To explore the feasibility of joint screening of the two primary immunodeficiency diseases [severe combined immunodeficiency (SCID) and X-linked agammaglobulinemia(XLA)] and spinal muscular atrophy(SMA) in newborns by multiplex real-time quantitative PCR technology, and to provide evidence for early screening, diagnosis and treatment of children.Methods:Cross-sectional study. From July 2021 to January 2023, a total of 103 240 dry blood spots samples of newborns were collected which were delivered to Neonatal Disease Screening Center of Zhejiang by cold chain transportation. The concentrations of the T cell receptor excision ring (TREC), Kappa deletion of the recombinant excision loop (KREC), and exon 7 deletion of Survival Motor Neuron 1 (SMN1) gene in dry blood spots were simultaneously detected by multiplex real-time fluorescence quantitative PCR, taken ribonuclease P/MRP 30 000 subunits (RPP30) as an internal reference gene. The positive newborns were further diagnosed by other laboratory tests and gene sequencing was taken as gold standard. Children samples from 1 case of SCID, 3 cases of XLA and 2 cases of SMA were used for positive verification. The correlation between detected concentration of TREC/KREC and basic information in newborns were analyzed. The differences among groups for each factor were analyzed.Results:One case of SCID, 2 cases of XLA, 9 cases of SMA and 7 cases of other genetic diseases (4 cases of DiGeorge syndrome, 1 case of trisomy 21 syndrome, 1 case of Noonan syndrome and 1 case of super male syndrome) were identified by multiplex real-time fluorescence quantitative PCR. The positive predictive values of screening neonatal SCID, XLA and SMA were 2.44% (1/41), 2.78% (2/72) and 9/9 respectively. Taking the samples from clinically diagnosed 1 case of SCID, 3 cases of XLA and 2 cases of SMA as positive validation samples, which were all identified. The detected results of TREC/KREC correlated with time of blood collection, sex, weight, gestational age and delivery mode of newborns, whose r values were 0.162/0.187, 0.066/0.032, 0.045/0.042, ?0.015/?0.088 and 0.014/0.068 respectively (all P<0.05). Conclusions:Relying on current neonatal screening platform in Zhejiang, it is feasible to screen jointly two kinds of primary immunodeficiency diseases and spinal muscular atrophy in newborns by multiple real-time fluorescence quantitative PCR technology.

Objective:To investigate the clinical effects of adjustable external fixation traction combined with arthroscopic microfracture in the treatment of osteochondral lesions of the talus (OLT).Methods:A retrospective study was conducted to analyze the data of 27 OLT patients who had been treated at Department of Orthopedics, Beijing Rehabilitation Hospital from May 2017 to March 2022. There were 16 males and 11 females, aged (32.4±7.2) years. Lesion site: 23 medial and 4 lateral cases; Hepple staging: 7 cases at stage Ⅰ, 15 cases at stage Ⅱ, and 5 cases at stage Ⅲ; disease duration: (10.6±3.3) months. All the patients were treated by adjustable external fixation traction combined with arthroscopic microfracture. Recorded were the patients' visual analogue scale (VAS) pain scores and American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot scores at 6 months and 12 months after surgery, levels of interleukin-1 (IL-1), interleukin-6 (IL-6) and tumour necrosis factor- α (TNF- α) at 1 month after surgery, lesion area at 12 months after surgery, and incidence of complications. Results:The follow-up time for this cohort was (16.2±6.7) months. The AOFAS score was (61.52±6.75) points before surgery, (84.15±5.56) points at 6 months after surgery and (95.67±4.30) points at 12 months after surgery. The VAS score was (5.88±1.02) points before surgery, (2.12±0.48) points at 6 months after surgery and (0.66±0.36) points at 12 months after surgery. The two-by-two comparisons between the 3 time points for the above items were statistically significant ( P<0.05). IL-1 was (32.37±6.64) pg/mL, IL-6 (34.04±7.12) pg/mL, and TNF- α (17.89±4.96) ng/L at 1 month after surgery in the 27 patients, all of which were significantly lower than their preoperative levels [(96.63±14.80) pg/mL, (102.33±20.42) pg/mL, and (54.48±9.33) ng/L] ( P<0.05). The lesion area was (28.66±6.52) mm 2 at 12 months after surgery, significantly smaller than the value before surgery [(128.52±11.32) mm 2] ( P<0.05). Infection at the adjustable external fixation needle track occurred in 1 patient and lower limb thrombosis in 2 patients. Conclusion:In the treatment of OLT, adjustable external fixation and traction combined with arthroscopic microfracture can achieve satisfactory results and improve symptoms for the patients.

Advanced radiotherapy technology enables the dose to more accurately conform to the tumor target area of the patient, providing accurate treatment for the patient, but the gradient of the patient's radiation dose at the tumor edge is getting larger, which putting forward higher requirements for radiotherapy dose verification. The dose verification system software KylinRay-Dose4D can verify the patient's pre-treatment plan and the in vivo/on-line dose during the patient's treatment, providing important reference for the physicist to modify the radiotherapy plan and ensuring that the patient receives accurate treatment. This study introduces the overall design and key technologies of KylinRay-Dose4D, and tests the pre-treatment plan dose checking calculation and 2D/3D dose verification through clinical cases. The test results showed that the 2D/3D gamma pass rate (3 mm/3%) of KylinRay-Dose4D reconstructed dose compared with TPS plan dose and measured dose is larger than 95%, which indicating that the reconstructed dose of KylinRay-Dose4D meets the requirement of clinical application.
Humans ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted/methods* ; Radiotherapy, Intensity-Modulated/methods* ; Software ; Neoplasms ; Phantoms, Imaging ; Radiometry/methods*

Objective:To investigate the clinical effects of tibial transverse bone transport assisted by nose ring drainage (NRD) in the treatment of foot and ankle chronic osteomyelitis.Methods:A retrospective study was conducted to analyze the data of 32 patients with foot and ankle chronic osteomyelitis who had been treated at Department of Orthopedics, Beijing Rehabilitation Hospital from March 2013 to February 2022. The patients were assigned into a study group and a control group. In the control group, there were 12 males and 3 females, aged (39.5±8.8) years. The osteomyelitis was located at the distal tibia in 4 cases, at the talus in 1 case, at the calcaneus in 2 cases, at the midfoot in 4 cases and at the forefoot in 4 cases. According to the Crerny-Mader classification, there were 5 cases of type Ⅰ and 10 cases of type Ⅱ. The control group was treated with Ilizarov transverse tibial bone transport in combination with thorough debridement and anti-infection therapy. In the study group, there were 12 males and 5 females, aged (42.3±13.4) years. The osteomyelitis was located at the distal tibia in 4 cases, at the talus in 1 case, at the calcaneus in 3 cases, at the midfoot in 5 cases and at the forefoot in 4 cases. According to the Crerny-Mader classification, there were 7 cases of type Ⅰ and 10 cases of type Ⅱ. The study group was treated with NRD drainage in addition to the procedures in the control group. The curative effects were evaluated by comparing the cure, recurrence, amputation (toe amputation), antibiotic use time, wound healing time, relevant inflammatory indicators [WBC, neutrophil percentage (NEU), erythrocyte sedimentation rate (ESR), procalcitonin, interleukin-6 (IL-6), C-reactive protein (CRP)], and postoperative functional recovery between the 2 groups.Results:The 2 groups were comparable because there was no statistically significant difference in the general data between them ( P>0.05). The follow-up period was (36.0±9.8) months. There was no significant difference between the 2 groups in the cure or amputation (toe amputation) ( P>0.05), but the recurrence rate in the study group was significantly lower than that in the control group ( P<0.05). The antibiotic use time [(20.7±3.6) d] and wound healing time [(88.3±17.1) d] in the study group were significantly shorter than those [(37.9±6.5) d and (102.2±22.6) d] in the control group ( P<0.05). The ESR, IL-6 and CRP at 1 week after operation in the study group were significantly lower than those in the control group ( P<0.05), but there was no significant difference between the 2 groups in the WBC, NEU or PCT at 1 week after operation ( P>0.05). There was no significant difference either in the inflammatory indicators between the 2 groups at 1 month after operation ( P>0.05). In all patients, the inflammatory indicators like WBC, NEU, ESR, PCT, IL-6 and CRP at 1 week and 1 month after operation were significantly better than those before operation, and the Maryland foot functional score at 12 months after operation was significantly higher than the preoperative one ( P<0.05). There was no significant difference between the 2 groups in the Maryland foot functional score at 12 months after operation ( P>0.05). Conclusion:In the treatment of foot and ankle chronic osteomyelitis, compared with the Ilizarov transverse tibial bone transport, our tibial transverse bone transport assisted by NRD can achieve satisfactory therapeutic effects, shorten antibiotic use time and treatment cycle, and reduce recurrence rate.

OBJECTIVES: To analyze the results of neonatal screening for congenital hypothyroidism (CH) and hyperphenylalaninemia (HPA) in Zhejiang province from 1999 to 2022. METHODS: A total of 11 922 318 newborns were screened from September 1999 and December 2022 in Zhejiang province. The blood thyroid stimulating hormone (TSH) levels were measured by a fluorescence method and blood phenylalanine (Phe) levels were measured by fluorescence method or tandem mass spectrometry. TSH≥9 μIU/mL was considered positive for CH, while Phe>120 μmol/L and/or Phe/Tyr ratio>2.0 were considered positive for HPA. The positive newborns in screening were recalled, and the gene variations were detected by high-throughput sequencing and MassARRAY tests. RESULTS: The overall neonatal screening rate during 1999-2022 was 89.41% (11 922 318/13 333 929) and the screening rate was increased from 6.46% in 1999 to 100.0% in 2022. A total of 8924 cases of CH were diagnosed among screened newborns with an incidence rate of 1/1336. A total of 563 cases of HPA were diagnosed, including 508 cases of classic phenylketonuria (cPKU) and 55 cases of tetrahydrobiopterin deficiency (BH4D), with an incidence rate of 1/21 176. Ninety-seven out of 8924 cases of CH underwent genetic analysis. Gene mutations were detected in 9 CH related genes, the highest frequency mutations were found in DUOX2 gene (69.0%) with c.3329G>A (p.R1110Q) (18.2%) and c.1588A>T (p.K530X) (17.3%) as the hotspot mutations. There were 81 PAH gene variants detected in a total of 250 cases of cPKU, and c728G>A (p.R243Q) (24.4%), c.721C>T (p.R241C) (15.0%) were the hotspot mutations. Meanwhile 7 novel variants in PAH gene were detected: c.107C>A (p.S36*), c.137G>T (p.G46V), c.148A>G(p.K50E), c.285C>T (p.I95I), c.843-10delTTCC, exon4-7del and c.1066-2A>G. There were 12 PTS gene variants detected in 36 cases of BH4D, and c.259C>T (p.P87S) (31.9%) was the hotspot mutation. CONCLUSIONS The incident of CH has increased from 1999 to 2022 in Zhejiang province, and it is higher than that of national and global levels; while the incidence of HPA is similar to the national average. DUOX2 gene variation is the most common in CH patients; c.728G>A (p.R243Q) is the hotspot mutation in cPKU patients, while c.259C>T (p.P87S) is the hotspot mutation in BH4D patients.
Humans ; Infant, Newborn ; Neonatal Screening ; Dual Oxidases ; Congenital Hypothyroidism/genetics* ; Phenylketonurias/genetics* ; Thyrotropin

Objective　To investigate the biological characteristics of the serine protease inhibitor 15 (RmS-15) protein of Rhipicephalus microplus, and to perform molecular cloning and prokaryotic expression. Methods RmS-15 gene was amplified and sequenced to construct a phylogenetic tree to understand its evolutionary relationships. Bioinformatic tools were used to analyze the physicochemical properties, signal peptide, secondary and tertiary structure of the RmS-15 protein. In addition, DNA star and ABCpred were used to predict potential B-lymphocyte antigenic epitopes of RmS-15 protein, and potential T-lymphocyte antigenic epitopes were predicted by SYF-PEITHI and IEDB. Finally, the RmS-15 recombinant protein of Rhipicephalus microplus was obtained using the E.coli prokaryotic expression system and purified. Results The RmS-15 gene with 1 212 bp was successfully amplified, encoding a protein comprising 403 amino acids. Phylogenetic tree results indicated high conservation of the RmS-15 protein among different tick amino acid sequences, with the closest phylogenetic relationships to the Rhipicephalus haemaphysaloides and the Rhipicephalus sanguineus. The results of physicochemical property analysis showed a 20-amino acid signal peptide at the N-terminus of the RmS-15 protein, with a relative molecular weight and theoretical isoelectric point of 44 000 and 6.68, respectively. The protein showed an average hydrophilicity of 0.001, classifying it as a stably hydrophilic protein. The results of antigenicity analysis showed that the dominant fragments of B-cell antigenic epitopes of RmS-15 protein were 102-112 aa,147-152 aa and 207-215 aa, and the dominant fragments of T-cell antigenic epitopes were 3-11 aa, 6-14 aa, 27-33 aa, 34-37 aa. Protein expression results showed that RmS-15 protein exhibited high expression levels in the supernatant after induction with 0.8 mmol/L IPTG at 16 ℃ for 24 hours and reached a successful purification with a single band of the protein molecular weight of 44 000. Conclusions The method of prokaryotic expression and purification of RmS-15 protein from Rhipicephalus microplus was successfully established. Bioinformatics analysis demonstrated its strong antigenicity, suggesting its potential to develop as a candidate vaccine for ticks.

Objective:To explor the effects of mentalization-based family therapy (MBFT) on the emotions of anxiety and depression, reflective functions, and related brain regions of adolescents with depression.Methods:Thirty-two adolescent patients with depression were selected in Beijing Huilongguan Hospital.Among them, fifteen patients treated with conventional drugs were defined as the control group, the other seventeen patients served as the observation group.On the basis of routine drug treatment, patients in the observation group were treated with 1.5 h MBFT once a week, for a total of 3 months.Among them, 10 patients underwent resting functional magnetic resonance imaging(rs-fMRI) scans before and after the intervention.SPSS 23.0 was used for statistical analysis of scale evaluation data by using chi-square test and repeated-measure analysis of variance, and pairwise Pearson correlation was used to construct a brain network matrix for the MRI results.The patient's brain network matrix was put into Gretna to calculate, and then use repeated measures analysis of variance and t test for judgment. Results:(1)Repeated measurement analysis of variance showed, over time and group, before and after intervention, the group×time interactions of patients' reflective function, anxiety, depression were statistically significant ( F=5.113, 9.350, 5.264, all P<0.05). Further simple effect analysis showed, in the observation group, there were statistically significant differences in patients' anxiety ((55.09±14.35) vs (30.58±3.62)), depression ((61.58±16.81) vs (31.00±3.48)) before and after intervention (all P<0.01). There was no significant difference in reflective function ((8.71±0.51) vs (8.87±0.50) ) in the observation group before and after intervention( P> 0.05). The reflective function((8.92±0.33) vs (8.73±0.35)) and anxiety((50.67±13.88) vs (45.78±12.89)) scores of the control group were not statistically significant different before and after intervention(all P>0.05), and the depression scores of the control group before and after inter-vention((69.33±9.11) vs (50.94±7.99)) were statistically significant different( P<0.01). (2)rs-fMRI showed that, using the posterior parietal lobe and the medial prefrontal lobe as seed points, the group×time interactions with right inferior frontal gyrus, right fusiform gyrus, right superior temporal gyrus, left middle temporal gyrus and right superior frontal gyrus were statistically significant ( F=29.56, 20.63, 8.69, 9.49, 8.62, all P<0.05). The further simple effect analysis showed in the observation group, the pre-test was less than the post-test, and the difference was statistically significant( P<0.05). In the control group, there were not statistically significant differences in the functional connection strength before and after the test(all P>0.05). (3)The brain network analysis of 264 regions of interest (ROIs) in the whole brain showed that the number of modules in the observation group was less than that in the control group, and the difference was statistically significant ( t=-2.64, P=0.046). Conclusion:MBFT can improve the anxiety and depression of adolescent patients with depression.It enhances the functional link between mentalization related brain regions and the whole brain, and enhances the activity of the whole brain link to become more integrated.

Objective:To analyze the related factors affecting the inter-arm systolic blood pressure difference (IASBPD) in a physical examination population.Methods:A total of 3 600 adults who underwent physical examination and completed the arteriosclerosis test in the first affiliated hospital of Anhui medical university from January 2019 to June 2021 were selected as the participants by systematic sampling method. Data on age, sex, and history of smoking, heavy drinking, hypertension, type 2 diabetes, and coronary heart disease were recorded. The height, weight, waist circumference, hip circumference, total muscle, total fat and body fat ratio were measured, and body mass index was calculated. The blood pressure of the limbs, ankle brachial index (ABI) were measured synchronously with the arteriosclerosis tester of the Chinese Academy of Sciences, and the IASBPD were calculated. According to the IASBPD value, the participants were divided into two groups: IASBPD<10 mmHg (1 mmHg=0.133 kPa) group and IASBPD≥10 mmHg group, The differences between the two groups were compared, and the related influencing factors of IASBPD were analyzed by multivariate logistic regression.Results:Weight, body mass index, waist circumference, hip circumference, systolic blood pressure, diastolic blood pressure, total muscle, total fat, body fat rate, history of hypertension, proportion of type 2 diabetes mellitus, and proportion of history of coronary heart disease in the IASBPD≥10 mmHg group was higher than that of IASBPD<10 mmHg group [(69.1±11.2) vs (65.3±10.8) kg, (25.6±3.4) vs (24.4±3.3) kg/m 2, (91.3±11.3) vs (87.8±10.6) cm, (98.5±10.4) vs (96.5±9.8) cm, (139.7±20.0) vs (129.7±17.6) mmHg, (80.3±11.6) vs (76.7±10.1) mmHg, (47.5±9.1) vs (45.3±8.8) kg, (19.4±7.0) vs (17.6±6.4) kg, (27.9%±8.5%) vs (26.8%±8.1%), 41.1% vs 29.3%, 16.6% vs 11.7%, 13.1% vs 7.3%] (all P<0.05); ABI was lower than that in IASBPD<10 mmHg group [(1.15±0.11) vs (1.20±0.09)] ( P<0.001). There were no significant differences in height, smoking history and heavy drinking history between the two groups (all P>0.05). Multivariate logistic regression analysis showed that age, systolic blood pressure, body weight and ABI were independent influencing factors of IASBPD≥10 mmHg. Age, systolic blood pressure and body weight were positively correlated with IASBPD≥10 mmHg, while ABI was negatively correlated with IASBPD≥10 mmHg. Conclusion:Increases in age, systolic blood pressure, and body weight and a decrease in ABI are important influencing factors leading to the elevation of IASBPD.

Objective:To systematically evaluate the efficacy and safety of obinutuzumab-based regimen versus rituximab-based regimen in treatment of B-cell non-Hodgkin lymphoma (B-NHL).Methods:The Cochrane clinical controlled trials database, PubMed, Embase, American Society of Hematology meeting proceedings, American Society of Clinical Oncology annual meeting proceedings and ClinicalTrails database were searched for studies on the use of regimens containing obinutuzumab or rituximab for the treatment of B-NHL. Patients were divided into obinutuzumab group and rituximab group according to their medication status. Review Manager 5.3 software was used to compare the efficacy and safety of the two groups.Results:A total of 7 randomized controlled trials were selected, including 4 235 patients (1 430 cases of follicular lymphoma, 2 102 cases of diffuse large B-cell lymphoma, and 703 cases of other B-NHL); 2 121 cases were in the obinutuzumab group and 2 114 cases were in the rituximab group. Among 4 162 patients who could be evaluated, the objective response rate (ORR) in the obinutuzumab group was higher than that in the rituximab group [75.1% (1 565/2 083) vs. 72.7% (1 512/2 079); OR = 1.19, 95% CI 1.01-1.41, P = 0.03]. Progression-free survival (PFS) in the obinutuzumab group was better than that in the rituximab group ( HR = 0.86, 95% CI 0.75-0.99, P = 0.03). Among 3 542 patients who could be evaluated for adverse reactions, the incidence of grade 3-4 adverse reactions in the otuzumab group was higher than that in the rituximab group [61.8% (1 098/ 1 776) vs. 54.2% (958/1 766); OR = 1.50, 95% CI 1.29-1.74, P < 0.001], the incidence of grade 3-4 infusive-related adverse reactions [7.5% (158/1 776) vs. 3.1% (65/1 766); OR = 2.56, 95% CI 1.91-3.45, P < 0.001] and neutropenia [34.1% (597/1 749) vs. 29.4% (511/1 738); OR = 1.27, 95% CI 1.09-1.47, P = 0.002] in the obinutuzumab group were higher than those in the rituximab group. Conclusions:The ORR and PFS of B-NHL patients treated with obinutuzumab-based regimen are better than those treated with rituximab-based regimen, but the influence of adverse reactions should be considered when selecting the regimen.