Objective:To explore the clinical phenotype and genetic etiology for a Chinese pedigree affected with Autosomal dominant polycystic kidney disease (ADPKD).Methods:A pedigree with ADPKD diagnosed at the Department of Gynaecology of the First Affiliated Hospital of Zhengzhou University in December 2020 was selected as the study subject. Clinical data of the pedigree was collected, and whole exome sequencing (WES) was carried out for the proband. Candidate variants were verified by Sanger sequencing of the proband and her relatives. This study was approved by the First Affiliated Hospital of Zhengzhou University (Ethics No. KS-2018-KY-36).Results:Fetal ultrasonography showed increased volume and parenchymal echogenicity in both kidneys. The fetus was found to harbor c. 11098C>T (p.R3700C) and c.11039T>C (p.F3680S) compound heterozygous variants of the PKD1 gene, which were respectively inherited from its mother and father. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), both variants were predicted to be likely pathogenic (PM1+ PM2_Supporting+ PP3). Conclusion:The c. 11098C>T (p.R3700C) and c. 11039T>C (p.F3680S) compound heterozygous variants of the PKD1 gene probably underlay the ADPKD in the fetus. Above finding has provided guidance for the genetic counseling and prenatal diagnosis for this pedigree.

Abstract: Objective　To retrospectively analyze the clinical characteristics of patients with Talaromyces marneffei infection (TSM) complicated by osteolytic lesions in non-HIV infected individuals, aiming to improve the diagnosis and treatment of TSM in non-HIV patients. Methods　Clinical characteristics and laboratory data, of non-HIV adults diagnosed with TSM complicated by osteolytic destruction were collected from the People's Hospital of Liuzhou City from January 2019 to October 2023, covering nearly five years. Results　All 14 patients were healthy hosts without HIV infection or any underlying diseases. All patients had systemic manifestations such as fever, anemia, and weight loss, while 7 patients experienced bone pain. The initial symptoms mainly included neck mass, ulceration, and purulent discharge from the skin. Laboratory tests showed elevated white blood cell counts, C-reactive protein, and erythrocyte sedimentation rate. The CD4+ lymphocyte count ranged from 157 to 1 477 cells/μL, and CD8+ lymphocyte counts from 112 to 1 779 cells/μL, with CD4+/CD8+ ratios from 0.651 to 3.797. Pathogenic evidence was obtained from positive cultures of purulent or ulcerative secretions from the neck, chest, face, and joints in 9 cases, positive metagenome sequencing of mediastinal lymph nodes in 1 case, positive cultures of sputum and bronchoalveolar lavage fluid in 4 cases, and positive blood cultures in 1 case. Imaging findings showed osteolytic destruction involving any part of the skeleton. Nine patients improved with treatment, 2 died, 3 showed no improvement, and 6 cases were misdiagnosed as lung cancer or metastatic tumors. After antifungal treatment, 7 patients developed non-tuberculous mycobacterial infection, and 4 cases were co-infected with TSM and Mycobacterium tuberculosis. Conclusions　TSM complicated by osteolytic destruction is more common in non-HIV infected, with prominent manifestations of severe bone or joint pain. Early diagnosis is difficult, and it is prone to be complicated by non-tuberculous mycobacterial or Mycobacterium tuberculosis infection, leading to frequent misdiagnosed as cancer and tumors clinically.

Tyrosol is a natural polyphenolic product that is widely used in chemical, pharmaceutical and food industries. Currently, the de novo synthesis of tyrosol by Escherichia coli suffers from issues such as low cell density and poor yield. Therefore, the phenylpyruvate decarboxylase mutant ARO10^F138L/D218G obtained in our previous study was fused with an alcohol dehydrogenase from different microorganisms for fusion expression, and the optimal ARO^{10F138L/D218G}-L-YahK produced 1.09 g/L tyrosol in shake flasks. In order to further improve tyrosol production, feaB, a key gene in the competing pathway of 4-hydroxyphenylacetic acid, was knocked out, and the resulted strain produced 1.26 g/L tyrosol with an increase of 21.15% compared to that of the control. To overcome the low cell density in tyrosol fermentation, the quorum-sensing circuit was used to dynamically regulate the tyrosol synthesis pathway, so as to alleviate the toxic effect of tyrosol on chassis cells and relieve the growth inhibition. Using this strategy, the yield of tyrosol was increased to 1.74 g/L, a 33.82% increase. In a 2 L fermenter, the production of tyrosol in the engineered strain TRFQ5 dynamically regulated by quorum-sensing reached 4.22 g/L with an OD₆₀₀ of 42.88. Compared with those in the engineered strain TRF5 statically regulated by induced expression, the yield was increased by 38.58% and the OD₆₀₀ was enhanced by 43.62%. The combination of blocking the competing pathway using gene knockout technology, and reducing the inhibitory effect of tyrosol toxicity on chassis cells through quorum-sensing dynamic regulation increased the production of tyrosol. This study may facilitate the biosynthesis of other chemicals with high toxicity.
Escherichia coli/genetics* ; Biological Products ; Bioreactors ; Fermentation

Ulcerative colitis (UC) manifests as an etiologically complicated and relapsing gastrointestinal disease. The enteric nervous system (ENS) plays a pivotal role in rectifying and orchestrating the inflammatory responses in gut tract. Berberine, an isoquinoline alkaloid, is known as its anti-inflammatory and therapeutic effects in experimental colitis. However, little research focused on its regulatory function on ENS. Therefore, we set out to explore the pathological role of neurogenic inflammation in UC and the modulating effects of berberine on neuro-immune interactions. Functional defects of enteric glial cells (EGCs), with decreased glial fibrillary acidic protein (GFAP) and increased substance P expression, were observed in DSS-induced murine UC. Administration of berberine can obviously ameliorate the disease severity and restore the mucosal barrier homeostasis of UC, closely accompanying by maintaining the residence of EGCs and attenuating inflammatory infiltrations and immune cells overactivation. , berberine showed direct protective effects on monoculture of EGCs, bone marrow-derived dendritic cells (BMDCs), T cells, and intestinal epithelial cells (IECs) in the simulated inflammatory conditions. Furthermore, berberine could modulate gut EGCs-IECs-immune cell interactions in the co-culture systems. In summary, our study indicated the EGCs-IECs-immune cell interactions might function as a crucial paradigm in mucosal inflammation and provided an infusive mechanism of berberine in regulating enteric neurogenic inflammation.

OBJECTIVE: explore the expression of miR-155-5p in Wilms tumor and its effect in regulating the proliferation, migration and apoptosis of Wilms tumor cells. METHODS: Specimens of tumor tissues and paired adjacent tissues were obtained from 40 patients with Wilms tumor for detection of the expression levels of miR-155-5p using RT-qPCR. Wilms tumor cell line G401 was transfected with miR-155-5p mimics and miR-155-5p inhibitor to induce miR-155-5p over-expression and its inhibition, respectively, and the changes in the cell proliferation, migration and apoptosis were assessed using cell counting kit-8 (CCK-8), wound healing assay and fl ow cytometry. RESULTS: RT-qPCR showed that the expression of miR-155-5p decreased significantly in Wilms tumor tissues as compared with normal kidney tissues and was significantly associated with TNM stage ( < 0.05). In G401 cells, over-expression of miR-155-5p significantly inhibited the cell proliferation and migration and promoted cell apoptosis ( < 0.05), and down-regulation of miR-155-5p obviously enhanced the proliferation and migration and suppressed apoptosis of the cells ( < 0.05). CONCLUSIONS miR-155-5p is down-regulated in Wilms tumor and its expression level is correlated with TNM stage. miR-155-5p participates in the progression of Wilms tumor by inhibiting the proliferation and migration and promoting apoptosis of the tumor cells, and may serve as a novel biomarker for diagnosis, therapy and prognostic evaluation of Wilms tumor.
Apoptosis ; Cell Movement ; Cell Proliferation ; Humans ; Kidney Neoplasms ; genetics ; MicroRNAs ; genetics ; Neoplasm Invasiveness ; Wilms Tumor ; genetics

We explored the improved method to prepare decellularized kidney scaffold and provide experimental basis for kidney tissue engineering and renal pathology and toxicology in vitro research. We perfused rat kidneys with PBS (group control) and prepared the decellularized kidney scaffolds with sodium dodecyl sulfate (SDS) (group S), Triton X-100 combined with SDS (group TS), and Triton X-100 combined with SDS after repeated freezing and thawing (group FTS) in different flow velocity. Meanwhile we measured their fluid distributions and vascular resistance. We examined the degree of decellularization of acellular scaffolds by HE, DAPI staining and DNA quantification. We examined the retention of main composition and structural integrity of decellularized scaffolds by Masson, PAS and immunohistochemical staining. We also detected the ultrastructure, cytotoxicity and the level of growth factor of the scaffolds by scanning electron microscope, MTT and ELISA, respectively. The results showed that the time of decellularization in group FTS was less than that in group S and TS. The vascular resistance of scaffolds decellularized at 10 mL/min flow velocity was lower. The fluid distribution in groups S, TS and FTS was different from that in control group. No residual cell was detected by HE and DAPI staining. DNA content was less than 50 ng/mg. Masson, PAS and immunohistochemical staining results showed that there was extracellular collagen, polysaccharide, type I collagen, type IV collagen, fibronectin and laminin in the decellularized scaffolds, and the scanning electron microscope result showed the scaffolds had the honeycomb structure. The cytotoxicity level of decellularized scaffolds was between grade 0 to 1. The level of VEGF, EGF, IGF-1 and PDGF-BB in group FTS were significantly higher than those in group S and TS. In concluding, combining freeze-thawing with perfusion can produce more ideal and effective whole organ decellularized scaffold of rat kidney, and make a foundation for the study of kidney tissue engineering and in vitro pathology and toxicology of kidney.
Animals ; Collagen ; Extracellular Matrix ; Freezing ; Kidney ; Perfusion ; Rats ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVE: To investigate the transcriptome profile of genital tubercles (GTs) in male SD rats and explore the mechanism of hypospadias induced by Di (2-ethylhexyl) phthalate (DEHP). METHODS: Forty time-pregnant SD rats were randomly divided into 4 equal groups, namely GD16 group and GD19 group (in which the male GTs were collected on gestation day[GD]16 and GD19 for RNA-seq, respectively), control group and DEHP exposure group (with administration of oil and 750 mg/kg DEHP by gavage from GD12 to GD19, respectively).In the control and DEHP exposure groups, the GTs were collected from the male fetuses on GD19.5, and scanning electron microscopy and HE staining were used to observe the morphological changes.The differentially expressed genes (DEGs) in the GTs were screened using lllumina HiSeq 2000 followed by GO and KEGG enrichment analyses to characterize the transcriptome profile.Immunofluorescence assay was performed to verify the DEGs (Mafb) identified by RNA-seq results.Immunofluorescence assay and Western blotting were used to examine the expression levels of Mafb in the penile tissue. RESULTS: A total of 1360 DEGs were detected in the GTs between GD16 group and GD19 group by RNA-seq.Among these genes, 797 were up-regulated and 563 were down-regulated.These DEGs were mainly enriched in the cell adhesion plaque signaling pathway, axon guidance signaling pathway, and extracellular matrix receptor signaling pathway.Compared with that in GD16 group, Mafb was significantly up-regulated in GD19 group, which was consistent with the sequencing results.Mafb and β-catenin were significantly down-regulated in DEHP-exposed group compared with the control group ( < 0.01). CONCLUSIONS Mafb expression increases progressively with the development of GTs in male SD rats.DEHP exposure causes significant down-regulation of Mafb and β-catenin, suggesting that β-catenin signaling pathway that affects Mafb is related to DEHP-induced hypospadias in SD rats.
Animals ; Diethylhexyl Phthalate ; Female ; Gene Expression Profiling ; Humans ; Hypospadias ; MafB Transcription Factor ; Male ; Oncogene Proteins ; Phthalic Acids ; Pregnancy ; Rats ; Rats, Sprague-Dawley

Objective To investigate the short-term efficacy of plasma exchange(PE),PE combined with double plasma molecule absorption system(PE+DPMAS) for treating severe hepatitis B(SHB).Methods The clinical data in 70 patients with SHB were retrospectively analyzed.The patients were divided into the PE group and PE+-DPMAS group according to different treatment modes.The clinical symptoms,liver function,coagulation function,blood routine,renal function and electrolytes changes,score decrease of model for end-stage liver disease(MELD) before and after treatment were compared between the two groups.Results After treatment,the alimentary tract symptoms were improved,the grade of hepatic encephalopathy was reduced and MELD score was decreased,but there was no statistically significant difference in the short-term effective rate between the two groups(P＞0.05).After treatment ALT,TBIL,RBC,Hb and PLT in the two groups were decreased significantly(P＜0.05);the ALB level in the PE+DPMAS group was decreased,while K+ and C1 were increased(P＜0.05);the PTA and ALB levels in the PE group were increased,while WBC was decreased in the PE group(P＜0.05).Conclusion The two kinds of treatment method PE and PE-+DP-MAS are effective in treating SHB.PE+-DPMAS can reduce the plasma usage and improve serum K+,Cl-levels;PE is superior to PE+DPMAS in the aspects of improving coagulation function and ALB level.

Objective To observe the changes of tongue and pulse parameters in the patients with chronic hepatitis B(CHB)after pegylated interferon alpha-2a(PEG-IFNα-2a)treatment,to investigate its value in evaluation clinical efficacy of PEG-IFNα-2a treatment.Methods 120 patients with CHB who confirmed to the standard received PEG-IFNα-2a antiviral therapy for 48 weeks,and followed up for 24 weeks.The tongue and pulse parameters were detected by DS01-A type digital tongue and pulse presentation analyzer.The changes of liver function,serum HBV markers,HBV DNA,tongue and pulse parameters were observed before and after treatment.Results 113 patients completed the course of treatment,46 cases received complete response(response rate 40.7%).The response rate of liver stagnation and spleen deficiency group was higher than that of blood stasis group(95%CI:0.010-0.677,P <0.05).The baseline of tongue and pulse parameters had no significant difference between response group and non response group(h3 /h1,t =1.799,P =0.074;h4 /h1,t =1.383,P =0.169;h5 /h1,t′=0.461,P >0.05;W/t,t′=0.688,P >0.05;R,t =1.317,P =0.190;G,t =0.346,P =0.729;B,t =1.720,P =0.088).After 48 weeks treat-ment,and followed up for 24 weeks,the tongue and pulse parameters of response group and non response group were compared with baseline,h3 /h1,h4 /h1 decreased,R value and G value increased,the differences were statistically sig-nificant(Response group:h3 /h1,t =3.004,P =0.003;h4 /h1,t =2.702,P =0.008;R,t′=2.258,P <0.05;G,t′=3.052,P <0.05.Non response group:h3 /h1,t =1.978,P =0.049;h4 /h1,t =2.487,P =0.014;R,t′=2.661,P <0.05;G,t′=2.318,P <0.05).But there were no significant differences between the response group and no response group after treatment(h3 /h1,t′=0.191,P >0.05;h4 /h1,t =0.390,P =0.697 2;h5 /h1,t′=0.957,P >0.05;W/t,t =0.149,P =0.881;R,t =1.343,P =0.181;G,t =0.994,P =0.322;B,t =0.565,P =0.572).Conclusion The changes of tongue and pulse parameters have improved after treatment with PEG-IFNαin patients with CHB. However,the value in predicting the efficacy of antiviral therapy may be limited.

OBJECTIVE:To explore the effects of inhaled budesonide on the efficacy and related indexes of patients with acute bronchitis. METHODS:102 patients with acute bronchitis were randomly divided into control group and observation group. Control group was given 100 mg/(kg·d) Cefotaxime sodium injection,adding into 150 ml 0.9% Sodium chloride injection intravenously by 2 times,as well as sedation,oxygen inhalation,rehydration,correcting acid-base balance and other conventional treatment;ob-servation group was additionally given 2 ml Inhaled budesonide suspension,twice a day. The treatment course for both groups was 7 d. Clinical efficacy,erythrocyte immune complex rosette(E-ICR),high-sensitivity C-reactive protein(hs-CRP),peak expiratory flow rate(PEF),forced vital capacity(FVC),1 second forced exhaled volume(FEV1),time of body temperature returned to nor-mal,cough disappearance time,rale disappearance time before and after treatment,and incidence of adverse reactions in 2 groups were observed. RESULTS:The total effective rate in observation group was significantly higher than control group,time of body temperature returned to normal, cough disappearance time and rale disappearance time were significantly shorter than control group,the differences were statistically significant(P<0.05). Before treatment,there were no significant differences in the E-ICR and hs-CRP levels,PEF,FVC and FEV1 between 2 groups(P>0.05). After treatment,E-ICR and hs-CRP levels were significant-ly lower than before,and observation group was lower than control group,PEF,FVC and FEV1 were significantly higher than be-fore,and observation group was higher than control group,the differences were statistically significant(P<0.05). And there were no adverse reactions during treatment. CONCLUSIONS:Based on conventional treatment,inhaled budesonide has obvious efficacy in the treatment of acute bronchitis,and it can reduce E-ICR and hs-CRP,improve pulmonary functions,with good safety.