Objective:To report the clinical manifestation and genetic characteristics of a case of de novo Huntington′s disease due to paternal intermediate alleles. Methods:Clinical data and imaging features of a middle-aged female, who complained of unstable walking without positive family history and was admitted to Xuanwu Hospital, Capital Medical University on September 20, 2022, were retrospectively analyzed. The serum samples of the patient and her parents were used to screen HTT gene dynamic mutation in accordance with the principle of informed consent and voluntary. And the relevant literatures were reviewed. Results:This is a 38-year-old female with progressive course, who presented as ataxia, involuntary movement at the end of extremities, dystonia, and cognitive impairment. Imaging results showed atrophy of bilateral caudate nuclei, as well as decreased glucose metabolism of bilateral caudate nuclei, putamen and partial cortex. Genetic testing showed the abnormal expansion of polymorphic trinucleotide (CAG) repeats in HTT gene and confirmed the diagnosis of Huntington′s disease. The CAG repeat length of the patient was 17/47 (pathopoiesis), of the father was 17/35 (intermediate alleles), and of the mother was 17/17 (normal). Conclusions:Paternal intermediate alleles may cause the first case of Huntington′s disease in a family. Importantly, HTT gene screening should be performed for the patient and parents when the diagnosis of Huntington′s disease is clinically possible despite negative family history, to prevent the misdiagnosis.

Objective:To analyse the pathogenic bacteria distribution and clinical characteristics of late-onset sepsis (LOS) among premature infants with gestational age less than 34 weeks in Henan Province.Methods:The clinical data of 6 590 premature infants admitted to 17 medical institutions in Henan Province from January 2019 to December 2020 were retrospectively analyzed. The gestational age of infants was less than 34 weeks and was admitted to the neonatal ward within 7 days after birth. SPSS 19.0 statistical software was used for data analysis.Results:Among 6 590 premature infants LOS developed in 751 cases (11.40%), of whom the diagnosis was confirmed in 276 cases (36.75%) and 475 cases (63.25%) were diagnosed clinically. The fatality rate related to LOS was 13.58%. There were significant differences in the incidence of LOS and infection-related mortality among infants with different gestational ages and body weights ( χ2=388.894 and 13.572, χ2=472.282 and 9.257, P<0.05 or <0.01). Among 276 children with confirmed LOS, 286 strains of pathogenic bacteria were isolated. Gram-negative bacteria were most prevalent (178 strains), accounting for 62.24% of all infections, followed by fungi (58 strains, 20.28%). Klebsiella pneumoniae was most frequently detected Gram-negative bacteria (117 strains, 40.91%), among which 32.48% (38/117) was carbapenem-resistant Klebsiella pneumoniae. The proportion of diagnosed sepsis, the proportion of catheterization, and the infection-related mortality of infants with LOS in tertiary hospitals were all higher than those in secondary hospitals ( χ2=6.212, 5.313 and 4.435, all P<0.05). The proportion of exclusive breastfeeding in secondary hospitals was lower than that in tertiary hospitals ( χ2=19.216, P<0.05). The time of antibacterial drug use before infection in specialized hospitals was longer than that in general hospitals ( χ2=3.276, P<0.05). Conclusion:The incidence of LOS among preterm infants in Henan Province is high, which was mainly caused by Gram-negative bacteria. The clinical characteristics of LOS caused by different pathogens and in different health institutions are different, the prevention and control strategy should be developed accordingly to reduce the incidence LOS of preterm premature infants.

Objective:To investigate changes in the bladder morphological structure and function and the expression of transforming growth factor-beta1 (TGF-β1) pathway-related proteins in the bilateral spinal nerve amputated neurogenic bladder(NB) rat.Methods:A total of 64 female SD rats were included, and 32 of them underwent bilateral spinal nerve L6+ S1 amputation to construct the NB model and the others were used as sham operation controls.Rats in both NB and control groups received bladder cystometry 3, 6, 12, 24 weeks after corresponding operation.Collagen fibers in their bladder tissues were detected by Masson staining and Sirius scarlet staining.TGF-β1, Smad2 and Smad6 proteins were checked by immunohistochemical staining.TGF-β1 receptor Ⅰ protein was measured by Western blot.Results:Bladders in the NB group were instable, with bladder leak point pressure(BLPP) and underactive voiding pressures.The basal pressure [(22.10±2.51), (18.20±1.52), (31.20±2.82), (41.10±3.41) cmH 2O(1 cmH 2O=0.098 kPa)] and bladder volume [(22.30±1.72), (49.10±5.54), (30.30±2.68), (13.50±1.52) mL] of the NB rats at 3, 6, 12 and 24 weeks were significantly higher than those of the sham operation controls[(3.51±0.45) cmH 2O and (0.52±0.04) mL], and the difference were significant(all P<0.05). The bladder size and thickness in the NB group firstly increased (3, 6 weeks) and then decreased (12, 24 weeks), but the bladder weight increased continuously.Masson staining showed disordered fibrous connective tissues, disintegrated layered bla-dder wall, hypertrophied smooth muscle tissues and deposited intramuscular collagen on the nerve-amputated bladder wall.Sirius scarlet staining suggested that 24 weeks after nerve amputation, collagen Ⅲ increased greatly, and the ratio of type Ⅲ/Ⅰ collagen fibers (3.14±0.71) was significantly higher than that in the sham group (0.88±0.21) ( t= 7.48, P<0.01). According to the immunohistochemical staining results, the expressions of TGF-1β and Smad2 increased while the pathway inhibitory protein Smad6 decreased with time in the NB group.Western blot showed that the expression of TGF-β1 receptor Ⅰ in the amputated bladder was 1.3 and 1.6 folds higher than that in the sham group 12 weeks and 24 weeks after operation( t=6.06, 14.45, all P<0.01). Conclusions:In NB rats with bilateral spinal nerve amputated, bladder contraction becomes paralysis, intravesical pressure increases, bladder normal structure disintegrates and the fibrosis pathway TGF-β1/Smads is activated.Therefore, the key step of development of pediatric NB is bladder fibrosis, which should be prevented as early as possibly in the clinical practice.

Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis and neuromyelitis optica spectrum disorders (NMOSD) are central nervous system diseases mediated by autoimmune antibodies. With the improvement of diagnosis and treatment, the reports of the two diseases appearing in the same patient are increasing. To strengthen the understanding of this kind of comorbidity, two cases of anti-NMDAR-encephalitis coexisting with NMOSD admitted in our hospital were reported. The clinical manifestations and imaging examination of the two patients showed the evidence of involvement of both brain and spinal cord. The anti-NMDAR antibody and anti-aquaporin 4 antibody tests were positive. In addition, gamma globulin or corticosteroid impulse therapy was effective in the treatment of two patients.

Objective To assess whether AECOPD patients can breathe independently at the PIC window and thus whether NPPV was necessary after extubation.Methods We performed a prospective observational study, we used the spontaneous breathing trial (SBT)to assess whether each patient could breathe independently at the PIC window,then performed extubation.Patients who passed the SBT received oxygen therapy only,whereas those who failed received NPPV.However,if the former showed respiratory distress,they also received NPPV.The primary out-come variables were SBT pass/fail,the demand for NPPV and rate of reintubation within 72h following extubation. Results In all,23 patients were enrolled,15cases(65.2%)of which passed the SBT.Of these,12cases (80.0%) patients developed respiratory distress after extubation and required NPPV (one of whom required reintubation).Of the eight patients that failed,one received reintubation after NPPV.The reintubation rates within 72h following extuba-tion of SBT-pass(7.0%)and SBT-fail (13.0%)(χ2 =1.476,P>0.05)patients were comparable.Conclusion Most AECOPD patients can breathe independently at the PIC window,but nonetheless develop respiratory distress and thus require NPPV following extubation.

ObjectiveTo evaluate the improvement on test performance of UniCel DxH 800 automated hematology analyzer for complete blood count (CBC) by detecting its performance indicators and comparing the differences of the results with LH 750 hematology analyzer and ADVIA 2120 hematology analyzer.MethodsThe precision,carryover and linearity of UniCel DxH 800 in measurement of CBC were evaluated by using fresh blood samples and instrument quality control of products.To evaluate the accuracy of leukocyte differential count and reticulocyte count with the microscopic method as the “gold standard”.To calculate the bias and correlation between the results measured by LH 750,ADVIA 2120 and UniCel DxH 800 hematology analyzers and compare these three instruments on the validity of the alarm in abnormal cells.ResultsIntra-precision:The coefficient of variation (CV) of the results of RBC,Hb and MCV were less than 0.5％,the CV of WBC and PLT results were less than 1.5％.Inter-precision:the CV of the parameters above were less than 2.5％.The carryover rate of WBC,RBC,Hb,MCV and PLT were less than 0.51％.In the concentration range covered by clinical samples,the correlation coefficients between the measured values and theoretical value in testing WBC,RBC,Hb and PLT were greater than 0.999 ( P ＜0.01 ).The measurement results of WBC,RBC,Hb,MCV and PLT hy UniCel DxH 800,ADV1A 2120 and LH 750 hematology analyzers have good correlation (r ＞ 0.973,P ＜ 0.01 ).Correlation of reticulocyte count between the UniCel DxH 800 hematology analyzer and microscolpic method was significant (r =0.920,P ＜0.01 ).Correlation of leukocyte differential count about the grauulocytes, lymphocytes and eosinophils between the UniCel DxH 800 hematology analyzer and microscopic method was good (r =0.914,0.900 and 0.725,P ＜0.01 ),followed by monocytes ( r =0.612,P ＜0.01 ),which were better than the LH 750 with similar detection principle.The UniCel DxH 800 hematology analyzer demonstrated higher sensitivity (96.6％ ) for the alarm of abnormal cells and achieved a lower false-negative rate (2.5％ ).Meanwhile,the sensitivity of the neutrophil nuclei left shift was higher (90.5％ ) and the false-negative rate (5.0％) was lower.ConclusionsThe UniCel DxH 800 hematology analyzer for complete blood count shows advantages of high precision,low carryover rate and wide linear range.The results detected by the UniCel DxH 800 hematology analyzer have good correlation with the LH 750 and ADVIA 2120.

Objective To investigate the left ventricular remodeling (LVR) patterns and its relationships with cardiovascular risk factors in patients with obstructive sleep apnea syndrome (OSAS).Methods 148 patients with habitual snoring were diagnosed OSAS by polysomnography with AHI≥5 /h and blood pressure was monitored at same time.The clinical characteristics were collected and echocardiography was done next morning.LVR patterns was categorized as normal geometry (NG),concentric remodeling (CR),eccentric hypertrophy (EH) and concentric hypertrophy (CH) on the basis of RWT ＞0.42 and LVMI ＞46.7 g/m2.7 (female) or 49.2 g/m2.7 (male).Multinomial logistic regression was used to analysis the correlation of cardiovascular risk factors and LVR patterns.Results ①Of 148 OSAS patients,there were 115 (77.7％) with abnormal LVR,NG 33 (22.3％),CR 35 (23.6％),EH 37 (25％)and CH 43 (29.1 ％).②General clinical characteristics:Compared with NG group,BMI and waistline were increased in EH group (P ＜0.05 or P ＜0.01) ; Age,BMI,waistline,hypertension,dSBP and nSBP were increased in CH group (P ＜0.05 or P ＜0.01).Compared with CR group,male,age,BMI and waistline were increased in EH group (P ＜0.05 or P ＜0.01).Male,age,BMI,waistline,hypertension,nSBP and TG were increased in CH group (P ＜ 0.05 or P ＜ 0.01); Compared with EH group,the prevalence of hypertension was increased in CH group (P ＜0.05); Compared with EH group,the prevalence of hypertension was increased in CH group (P＜0.05).③Polysomnography parameters:Compared with NG group,AHI and ODI were increased in CR group (P＜0.05 or P＜0.01).AHI,ODI and T90 were increased,while Mean SaO2 and Lowest SaO2 were decreased in CH group (P ＜0.05 or P ＜ 0.01);Compared with CR group,AHI was decreased in EH group (P ＜0.05).Mean SaO2 was decreased,but T90 was increased in CH group (P＜0.05 or P ＜0.01); Compared with EH group,Lowest SaO2 was decreased in CH group (P＜0.05).④Blood pressure:Compared with NG group,4:00 SBP and 6:00 SBP were increased in CH group (P＜0.05) ;Compared with CR group,22:00 SBP was increased in CH group (P ＜0.05).⑤ In multinomial logistic regression analysis,after adjusting the confounding factors,CR was associated with AHI [odds ratio (OR) 1.035,P =0.024.EH was associated with Age (OR 1.094,P =0.016),BMI (OR 1.397,P =0.011) and 4:00 SBP (OR 1.124,P =0.026).CH was associated with Age (OR 1.084,P =0.028).Conclusions OSAS could cause result in LVR and the prevalence of four patterns were similar..Age and AHI were significant determinants of CH and CR respectively.Age,BMI and 4:00 SBP were significant determinants of EH.

Objective To detect changes of regional grey matter and white matter volume in patients of neuromyelitis optica (NMO) by voxel-based morphometry (VBM),and investigate its relationship with clinical variables.Methods Conventional magnetic resonance imaging (MRI) and structural threedimensional MRI were obtained from 20 NMO and 20 sex-and age-matched healthy volunteers.The comparison of grey matter and white matter volume between the two groups was analyzed by VBM tools of statistical parametric mapping (SPM) 5.Pearson correlational analysis was used to assess correlations between regional volume decrease and disease duration and expanded disability status scale (EDSS) scores in NMO patients.Results Compared with normal controls,NMO patients had grey matter atrophy in several cortical regions,such as right inferior frontal gyrus (cluster size 514),left superior temporal gyrus (282),right middle temporal gyvus (229) and right insula (211) (t =3.58-5.11,AlphaSim corrected,P ＜0.05).White matter atrophy was found in several subcortical regions in NMO patients,such as right precentral and postcentral gyrus (cluster size 457,110),left middle frontal gyrus (285),and right inferior parietal lobule (231) (t =2.90-4.25,AlphaSim corrected,P ＜ 0.05).Grey matter and white matter volume loss were not significantly correlated with clinical duration or EDSS score in NMO.Conclusion By means of VBM,regional atrophy of grey matter and white matter is found in NMO patients,which may provide evidence for brain structural abnormality in NMO.

Objective To explore the clinical features and diagnostic method of primary natural killer( NK)/T cell meningeal lymphoma. Methods An unusual case of a 19-year-old male with primary NK/T cell meningeal lymphoma was reported. His clinical presentation and laboratory findings were discussed. The related literatures have been reviewed. Results The patient presented with diplopia,headache, vomiting and facial drooping at the onset, followed by progressive pain and weakness of the four limbs. Cerebrospinal fluid showed significant increase in pressure, leukocytes number, levels of protein,normal glucose and adenosine deaminase, negative tuberculosis antibody and sterile staining. In cerebrospinal fluid cytological analysis, May-Grunwald-Gimsa staining showed large number of atypical lymphocytes with irregular nucleus and nuclear fission, Ki-67 immunostaining showed extensive proliferative activity of the lymphoid cells. Flow cytometric immunophenotypic analysis of cerebrospinal fluid indicated 97. 98 percent of cells expressed surface CD3, CD7, CD56, CD2, CD5, and partially expressed CD8. This was a rare immunophenotype for NK/T-cell. Cranial MRI with gadolinium showed thickening of the trigeminal nerve with slight enhancement and diffuse leptomeningeal enhancement. CT of the chest and abdomen and bone marrow biopsies were negative. He was diagnosed as primary NK/T cell meningeal lymphoma based on the clinical features and related examination. Conclusions Primary NK/T cell meningeal lymphoma is a rare type of primary central nervous lymphomas which has special immunophenotype. The clinical features include progressive raised intra-cranial pressure, multiple cranial and spinal nerve involvements. Cerebrospinal fluid cytological analysis and flow cytometric immunophenotypic analysis are key work-up for diagnosis. It has poor response to chemotherapy and radiotherapy.

Interleukin-21 is a type I cytokine mainly produced by activated CD4+ T cells that acts as a regulator of immune system. In this work, hIL-21cDNA was amplified from human peripheral blood lymphocytes by RT-PCR, and then inserted into pPIC9K. The recombinant vector pPIC9K-hIL21cDNA was linearized by Sac I, and transformed into Pichia pastoris strain GS115 by electroporation. Transformants were selected by G418 and confirmed by PCR. The recombinant protein was expressed and secreted into the supernatant after inducing by methanol. SDS-PAGE analysis indicated the molecular weight of rhIL-21 was about 16 kD. ELISA results show that the yield of rhIL-21 reach 229.28 mg/L, rhIL-21 was purified from culture supernatants, and it was purified to about 95% purity with ion-exchange chromatography. When co-stimulate with Con A, rhIL-21 can promote the proliferation of human lymphocytes. This is the first expression of bio-active rhIL-21 in Pichia pastoris. It lays a foundation for further research in immunotherapy and cancer therapy.
Electroporation ; Genetic Vectors ; genetics ; Humans ; Interleukins ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; analysis ; biosynthesis ; genetics