Objective To investigate the therapeutic effect of a combination of artemether and daphnetin against Plasmodium berghei ANKA strain in mice. Methods Groups of P. berghei infected mice were treated with various oral doses of artemether and daphnetin according to “4-day suppress assay”. Thin blood smears were made on the fifth day after inoculation of parasites and the parasitemia reduction rate was calculated. The ED 50 values obtained were plotted on isobolograms. A combined action of artemether and daphnetin was assessed. Results Artemether 0.4mg/(kg?d)?4d exhibited no detectable antimalarial effect, while artemether 0.4mg/(kg?d)?4d combined with daphnetin 7.7 mg/kg.d?4d showed potent antiparasile efficacy. The ED 50s of artemether in combination with daphnetin were lower than that of single artemether or daphnetin. The R-values were higher than 0.4, but lower than 2.7. Conclusion The combination of artemether with daphnetin showed an additive antiparasile effect.

Objective To analyze the test results of persons with noise -induced hearing loss using disyllabic Mandarin speech test materials (MSTMs) .Methods 57 subjects with noise -induced hearing loss (noise group) were divided into three groups according to the duration of noise exposure :less than 10 years ,between 11 and 20 years and more than 20 years ,and also separated into 2 groups in terms of the degrees of hearing loss (group A :pure tone average thresholds of 0 .5 ,1 ,2 ,and 4 kHz less than or equal to 30 dB HL ,group B:pure tone average threshold of 0 .5 ,1 ,2 ,and 4 kHz greater than 30 dB HL) .66 subjects with normal hearing were in the control group .All the subjects enrolled in this study could speak Mandarin well in their daily life .Nine lists of disyllabic Mandarin speech test materials were utilized to test SRT and P -I function for these groups respectively .Results PTA and SRT in the noise group were much higher than the control group ,the average slope of P -I function was 5 .94% /dB ,and the curve of P-I function moved to the right side .SRT showed obvious changes as the increase of the years of noise exposure and especially in the group which had more than 20 years noise exposure (P<0 .01) . SRT of group B which had more hearing loss were greater than that of group A (P<0 .01) .Conclusion SRTs in the noise group are much higher than the control group with the average slope of P -I functions decreasing and the curve moving to the right side .As the years of noise exposure increase and the degrees of hearing loss become more severe ,SRT rise more apparently .

OBJECTIVETo investigate the methods and solve the technical bottlenecks in the preparation of recombinant human protein hZP3 using the baculovirus expression system and pave the technical ground for the production and application of recombinant hZP3.

METHODSThe recombinant vector pFASTBAC HTa-hZP3 was constructed and transferred to competent E. coli cells carrying bacmid to produce recombinant bacmid by homologous recombination. Sf9 cells were transfected with the recombinant bacmid to produce recombinant baculovirus. Full-length recombinant hZP3 (amino acids 1-424) and truncated recombinant hZP3 (amino acids 23-348) were expressed in the sf9 cells by infection with the recombinant baculovirus. The expression time of hZP3 was determined by Western blot and its purification was explored.

RESULTSThe recombinant bacmid and baculovirus were successfully constructed for expressing both the full-length and truncated hZP3. The maximal expression of recombinant hZP3 in the sf9 cells was achieved at 72-96 hours after baculovirus infection. Some of the recombinant hZP3 with His-tag could bind affinity matrix and got purified but most of the solubilized hZP3 passed through and the reasons remained unknown. Purified recombinant hZP3 labeled with Dylight Dye488 was able to bind human sperm.

CONCLUSIONIt is feasible to express recombinant hZP3 in insect cells using the baculovirus system though the yield of hZP3 needs to be optimized. The methods for efficient enrichment and purification of recombinant hZP3 require further exploration.

Baculoviridae ; genetics ; metabolism ; Blotting, Western ; Egg Proteins ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Membrane Glycoproteins ; genetics ; metabolism ; Receptors, Cell Surface ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; methods ; Zona Pellucida Glycoproteins

OBJECTIVE: To valuate test results of normal hearing persons with different ages using disyllabic mandarin speech test materials (MSTMs). Obtaining speech recognition threshold (SRT) and P-I function of different ages as clinical reference of hearing recovery and individual's ability to perceive and process speech. METHOD: One hundred and twenty subjects with normal hearing who speak mandarin well in their daily lives were enrolled in this study and divided into four groups (18-30, 31-40, 41-50 and 51-60 years old). Nine lists of disyllabic mandarin speech test materials with equal difficulty were utilized to test each age group. RESULT: There are good agreement between SRT and mean PTA thresholds (at 0.5, 1.0, 2.0, and 4.0 kHz) in each age group. PTA thresholds have little change as age increasing in age group 18-30, 31-40 and 41-50 (P > 0.05). PTA threshold of age group 51-60 increases more apparently compared with the other three groups (P < 0.01). SRT thresholds of each age group increase as age increasing (P < 0.05) and SRT threshold of age group 51-60 increases more apparently (P < 0.01). Slopes of P-I function in each age group are 5.8%/dB, 4.7%/dB, 3.8%/dB, 2.9%/dB respectively. CONCLUSION Nine lists of disyllabic MSTMs were used to test normal hearing persons in different ages and SRT and P-I function of four different age groups were obtained. As age increases, SRT increases and slope of P-1 functions decreases. PTA and SRT thresholds of age group 51-60 increase more apparently. The SRT and P-I functions provide reference data of normal hearing for utilizing of disyllabic mandarin speech test materials clinically.
Adolescent ; Adult ; Female ; Humans ; Language ; Male ; Middle Aged ; Phonetics ; Reference Values ; Speech Reception Threshold Test ; Young Adult

Background/Aims: Anti-phospholipase A2 receptor (PLA2R) autoantibody is the main biomarker of idiopathic membranous nephropathy (IMN). We aimed to find a new cutoff value of anti-PLA2R for patients with IMN and to explore the relevance between this antibody and baseline clinical parameters. Methods: A total of 670 subjects including 374 IMN cases and 296 non-IMN controls were included between January 2017 and January 2020. All clinical parameters were collected at the time of renal biopsy. The levels of anti-PLA2R were detected by a commercial enzyme-linked immunosorbent assay (ELISA) kit. The optimal cutoff value was calculated by a receiver operating characteristic curve and compared in diagnostic efficiency. Results: The optimal cutoff value of anti-PLA2R for IMN was 7.45 RU/mL with the highest Youden index, and the corresponding sensitivity, specificity, positive predictive value and negative predictive value were 80.75%, 97.97%, 98.05% and 80.11%, respectively. Anti-PLA2R levels in IMN patients demonstrated a significant positive correlation with serum creatinine and 24-hour urinary protein, while they showed a negative correlation with serum albumin and estimated glomerular filtration rate. Conclusions The recommended cutoff value of anti-PLA2R is 7.45 RU/mL using ELISA detection for distinguishing IMN from non-IMN nephropathy. The level of anti-PLA2R is related to baseline renal function in IMN. This new threshold can improve the diagnostic efficiency and facilitate early diagnosis of IMN.

AIMTo identify genes that are differentially expressed in omental fat of normal weight subjects, obese subjects and obese type 2 diabetic patients.

METHODSUsing a home-made high-density cDNA microarray, we compared gene expression profile of omental fat from normal weigh subjects, obese subjects and obese type 2 diabetic patients, to identify adipose-specific genes associated with obesity and diabetes.

RESULTS119 and 257 genes were up-regulated in obese patients and obese diabetic patients respectively, while 46 and 58 genes were down-regulated in obese patients and obese diabetic patients respectively. 77 genes, including metabolism related genes (PDK4), and caveolin 2, metallo thionein 1B, were up-regulated in both obese and obese diabetic patients, while 8 genes, including key enzymes in lipid synthesis, such as HMG-CoA synthase, fatty acid synthase and stearoyl-CoA desaturase, were down-regulated in both groups. Another interesting finding was that tyrosine-3-monooxygenase/ tryptophan 5-monooxygenase activation protein theta (YWHAZ), a negative regulator for insulin signal transduction, was up-regulated only in obese diabetic patient, but not in normal-glycemic obese subjects.

CONCLUSIONOur study demonstrated that decrease of lipogenesis along with increase of fatty acids oxidation of adipose tissue could be a common cause of insulin resistance in obesity and type 2 diabetes, while functional changes of other genes, such as immune regulation genes,might also be involved in the pathogenesis of obesity and type 2 diabetes. Block of insulin signal transduction might trigger the transition from obesity to diabetes. Further exploration of these genes will greatly help us in the understanding of the pathogenesis of obesity and type 2 diabetes.

Abdominal Fat ; metabolism ; Adipose Tissue ; metabolism ; Adult ; Diabetes Mellitus, Type 2 ; genetics ; metabolism ; Female ; Gene Expression Profiling ; Humans ; Insulin ; metabolism ; Insulin Resistance ; Male ; Middle Aged ; Obesity ; genetics ; metabolism ; Oligonucleotide Array Sequence Analysis

Objective:To prepare human decellular adipose tissue matrix (DAT) as injectable homogenate by a specially developed high-speed soft tissue homogenizer with controllable temperature, and to investigate its cellular compatibility and filling effect.Methods:From March 2019 to March 2021, DAT was obtained in the 940th Hospital from normal adipose tissue after decellular treatment. The DAT was mixed with normal saline at the rate of 1: 12. The specially developed high-speed soft tissue homogenizer with controllable temperature was used to conduct homogenization at 803×g for 10 min. The produced DAT homogenizer could pass through the 27G needle smoothly. DAT homogenate was observed under scanning electron microscope and its cytocompatibility was detected. Finally, granular fat, DAT homogenate and DAT homogenate + ADSCs were injected into the back of rats respectively, and the filling effect, angiogenesis ability and inflammatory infiltration were compared.Results:After decellular treatment, adipose tissue changed into DAT without cellular residue. The particle size of DAT homogenate was about (749.91±136.79) nm, which was prepared by the specially developed temperature controlled high-speed soft tissue homogenater. The adhesion rate was (92.16±1.00) %, and the A value increased with time. The cell growth was good, and the homogenate had no toxicity to the cells. In vivo experiment, the filling effect of DAT homogenate and DAT homogenate + ADSCs was significantly better than that of granulated fat group ( P<0.01). In the granulated fat group, a large number of adipocytes were necrotic and fused to form oil sacs, while DAT homogenate, DAT homogenate + ADSCs showed no obvious degradation, and some adipocytes were generated. The results of CD31 staining indicated that the number of microvessels in DAT homogenate group and DAT homogenate + ADSCs group was higher than that in granulated fat group ( P<0.01). The results of CD68 staining indicated that the inflammatory infiltration in DAT homogenate group and DAT homogenate + ADSCs group was lower than that in granule fat group ( P<0.01). Conclusions:Using the self-developed temperature controlled high-speed soft tissue homogenate device, DAT could be prepared into DAT homogenate that can pass through 27G needle. It has good biocompatibility and filling advantages, and injective process is simple, and the trauma is small, and so it could be used as filling material.

Objective:To investigate the expression and clinical significance of vitamin D receptor (VDR) and human β-defensin-2(HBD2) in children with Helicobacter pylori (Hp) infection and gastritis.Methods:Eighty-one children who were hospitalized in the Department of Gastroenterology at Beijing Children′s Hospital from January 2021 to June 2023 and underwent endoscopic examination were collected.They were divided into Hp infection group and non-Hp infection group according to whether they were infected with Hp.The expression levels of VDR and HBD2 between two groups, and the correlation between VDR, HBD2 expression levels and gastritis were compared and analyzed.Results:Among 81 cases, 48 cases were Hp positive, including 24 males and 24 females, with an average age of (11.4±2.7) years; 33 cases were Hp negative, including 14 males and 19 females, with an average age of (11.3±2.6) years.There were no significant differences in sex and age between two groups ( P>0.05). The positive rates of VDR and HBD2 expression in gastric mucosa of children with Hp infection were higher than those of children without Hp infection, and the differences were statistically significant (87.5% vs.39.4%, 79.2% vs.63.6%, all P<0.05). The expressions of VDR and HBD2 had no correlation with age and sex ( P>0.05). The expressions of VDR and HBD2 were positively correlated with granular degeneration of gastric mucosa ( r=0.384, P<0.001; r=0.258, P=0.020). The expression of VDR was positively correlated with the degree of gastric inflammation ( r=0.365, P=0.001), while the expression of HBD2 was not correlated with the degree of gastric inflammation ( P>0.05). Conclusion:The expression levels of VDR and HBD2 in gastric mucosa of children infected with Hp are increased.The expression level of VDR is correlated with the degree of gastritis and the granular degeneration of gastric mucosa.The expression level of HBD2 is correlated with the granular degeneration of gastric mucosa.But there is no correlation between the level of HBD2 expression and the degree of gastritis.

Objective:To investigate long-term auditory changes and characteristics of Alport syndrome（AS） patients with different degrees of renal injury. Methods:Retrospectively analyzing clinical data of patients diagnosed AS from January 2007 to September 2022, including renal pathology, genetic detection and hearing examination. A long-term follow-up focusing on hearing and renal function was conducted. Results:This study included 70 AS patients, of which 33（25 males, 8 females, aged 3.4-27.8 years） were followed up, resulting in a loss rate of 52.9%.The follow-up period ranged from 1.1to 15.8 years, with 16 patients followed-up for over 10 years. During the follow-up, 10 patients presenting with hearing abnormalities at the time of diagnosis of AS had progressive hearing loss, and 3 patients with new hearing abnormalities were followed up, which appeared at 5-6 years of disease course. All of which were sensorineural deafness. While only 3 patients with hearing abnormalities among 13 patients received hearing aid intervention. Of these patients,7 developed end-stage renal disease（ESRD）, predominantly males （6/7）. The rate of long-term hearing loss was significantly different between ESRD group and non-ESRD group（P=0.013）. There was no correlation between the progression of renal disease and long-term hearing level（P>0.05）. kidney biopsies from 28 patients revealed varying degrees of podocyte lesion and uneven thickness of basement membrane. The severity of podocyte lesion was correlated with the rate of long-term hearing loss（P=0.048）, and there was no correlation with the severity of hearing loss（P>0.05）. Among 11 cases, theCOL4A5mutationwas most common （8 out of 11）, but there was no significant correlation between the mutation type and hearing phenotype（P>0.05）. Conclusion:AS patients exhibit progressive hearing loss with significant heterogeneity over the long-term.. THearing loss is more likely to occur 5-6 years into the disease course. Hearing abnormalities are closely related to renal disease status, kidney tissue pathology, and gene mutations, emphasizing the need for vigilant long-term hearing follow-up and early intervention.
Male ; Child ; Female ; Humans ; Nephritis, Hereditary/pathology* ; Retrospective Studies ; Kidney ; Deafness ; Hearing Loss/genetics* ; Kidney Failure, Chronic/pathology* ; Mutation