Objective:To investigate the clinical phenotype and genetic characteristics of patients with Fabry disease caused by a GLA variant, IVS4+919G>A.Methods:It was a prospective study. Fabry disease screening was conducted among high-risk population in Ninghai from October 2021 to August 2023. Those children with decreased α-galactosidase enzyme activity<2.40 μmol/(L·h) or elavated Lyso-GL-3 level>1.10 μg/L in dried blood spot (DBS) method underwent GLA genetic testing for diagnosis confirmation. Meanwhile, family screening was carried out. A proband and his family members diagnosed with Fabry disease were research subjects. The clinical and genetic characteristics of patients with Fabry disease caused by the GLA variant (IVS4+919G>A) were analyzed.Results:The female proband aged 9.8 years with pain in both lower limbs as the initial symptom was found to have a heterozygous GLA variant IVS4+919G>A among 102 patients. In family screening, there were 4 family members (proband's father, elder sister, elder male cousin and elder female cousin) with Fabry disease and a family member (proband's fifth aunt) with a GLA variant. Among these 4 diagnosed family members, the elder male cousin of the proband, a boy aged 13.2 years had a heterozygous GLA variant, IVS4+919G>A with intermittent pain in both lower limbs as the initial symptom. The proband′s father had knee joint pain. The proband′s elder sister had decreased vision and his elder female cousin had no obvious symptoms. The proband′s fifth aunt with a GLA variant had decreased vision.Conclusions:High-risk screening in children and family screening are helpful for early diagnosis and treatment of Fabry disease. Neuropathic pain may be a early symptom in children with Fabry disease caused by the GLA variant, IVS4+919G>A.

With various diseases ravaging internationally, the demands for recombinant adenoviral vector (Adv) vaccines have increased dramatically. To meet the demand for Adv vaccine, development of a new cell culture process is an effective strategy. Applying hyperosmotic stress in cells before virus infection could increase the yield of Adv in batch culture mode. Emerging perfusion culture can significantly increase the yield of Adv as well. Therefore, combining the hyperosmotic stress process with perfusion culture is expected to improve the yield of Adv at high cell density. In this study, a shake flask combined with a semi-perfusion culture was used as a scaled-down model for bioreactor perfusion culture. Media with osmotic pressure ranging from 300 to 405 mOsm were used to study the effect of hyperosmotic stress on cell growth and Adv production. The results showed that using a perfusion culture process with a hyperosmotic pressure medium (370 mOsm) during the cell growth phase and an isosmotic pressure medium (300 mOsm) during the virus production phase effectively increased the yield of Adv. This might be due to the increased expression of HSP70 protein during the late phases of virus replication. The Adv titer in a bioreactor with such a process reached 3.2×10¹⁰ IFU/mL, three times higher than that of the traditional perfusion culture process. More importantly, this is the first time that a strategy of combining the hyperosmotic stress process with perfusion culture is applied to the production of Adv in HEK 293 cells. It also reveals the reason why the hyperosmotic stress process increased the yield of Adv, which may facilitate the process optimization of for producing other Adv in HEK 293 cells.
Humans ; HEK293 Cells ; Genetic Vectors/genetics* ; Batch Cell Culture Techniques ; Bioreactors ; Perfusion

Objective To observe the impact factors of the diagnostic accuracy of fine needle aspiration biopsy(FNAB)for papillary thyroid carcinoma(PTC).Methods Totally 468 patients with single PTC confirmed by postoperative pathology who underwent FNAB before surgery were enrolled.The impact of clinica,l ultrasonic and pathological features on the accuracy of FNAB diagnosis were analyzed.Results The accuracy of FNAB for diagnosing PTC was 71.37%(334/468).The maximum diameter and location of PTC were both impact factors of the diagnostic accuracy of FNAB.The maximum diameter of 0.7 cm was the optimal cutoff value of FNAB for diagnosing PTC,and the diagnostic accuracy of FNAB for PTC with the maximum diameter＜0.7 cm and those≥0.7 cm was 62.96%(119/189)and 77.06%(215/279),respectively.The diagnostic accuracy of FNAB for PTC located in the difficult and easy area of puncture was 52.53%(52/99)and 76.42%(282/369),respectively.The diagnostic accuracy of FNAB for PTC with the maximum diameter≥0.7 cm and located in the easy area,≥0.7 cm and located in the difficult area,＜0.7 cm and located in the easy area,＜0.7 cm and located in the difficult area was 80.43%(185/230),61.22%(30/49),69.78%(97/139)and 44.00%(22/50),respectively.Conclusion The maximum diameter and location of PTC were both impact factors of the diagnostic accuracy of FNAB.

Primary acinic cell carcinoma (ACC) of the lung is extremely rare. The World Health Organization tumor classification defines ACC as "a malignant epithelial neoplasm that demonstrates some cytological differentiation towards (serous) acinar cells". It is considered to be a low-grade malignant tumor. Since the first case described by Fechner in 1972, less than 30 cases have been reported in the literature. The rarity of this tumor may leads it to be confused with other primary lung tumors and incorrectly diagnosed. We reported a female patient with primary ACC of the lung with mediastinum deviation at age of 27 years received a right pneumonectomy. She was followed up for 12 months postoperatively and remains well.

The purpose of this paper is to investigate the altered brain areas activated in functional magnetic resonance imaging before and after escitalopram antidepressant treatment in patients with major depressive disorder. Functional magnetic resonance studies showed abnormal activity in brain regions such as prefrontal cortex， cingulate cortex and striatum in patients prior to treatment， meantime， the abnormal activation of anterior cingulate， dorsolateral prefrontal and thalamus reflected the outcomes of escitalopram antidepressant therapy. Following the treatment with escitalopram， the fMRI activation of brain areas returned to normal， and those changes were significantly correlated with the improvement of depressive symptoms. So this paper provides a review of the brain functional activity changes in patients with major depressive disorder before treatment and at different time point of medication， as well as the correlation between altered brain functional activity and clinical symptoms.

Objective To study the efficacy and safety of intravesical chemotherapy combined with intravenous chemotherapy for high grade T1 (T1G3) bladder cancer after transurethral resection of bladder tumor(TURBT).Methods From January 2012 to December 2015,111 patients with high grade T1 (T1 G3) bladder cancer were retrospectively reviewed.Thirty-six patients received TURBT and intravesical chemotherapy and intravenous chemotherapy (group A),75 patients received TURBT and intravesical chemotherapy(group B).In group A,there were 28 males and 8 females,with average age 66.2 years;in group B,there were 59 males and 16 females,with mean age 67.9 years.There was no statistical difference between the two groups in age,sex,smoking history,tumor diameter,tumor number.1-year recurrence-free survival (RFS),1-year progression-free survival (PFS),intravenous chemotherapy adverse reaction were analyzed.Results All the patients were followed-up for 12 months.9 patients relapsed in group A,1-year RFS rate 75％,and the median RFS of the 9 patients was 9 (3-11) months.36 patients relapsed in group B,1-year RFS rate 52％,and the median RFS of the 36 patients was 7 (3-11) months.There was statistically significant difference between the two group(P =0.02).One patient progressed in group A,1-year PFS rate 97.2％,and the PFS was 9 months.Six patients progressed in group B,1-year PFS rate 92％,and the median PFS was 9.5(6-12) months.There was no statistically significant difference (P =0.305) between the two group.Only 1 case (3％) appeared Ⅲ° or above intravenous chemotherapy adverse reaction.Conclusions Intravesical chemotherapy combined with intravenous chemotherapy offers a better RFS rate than the intravesical chemotherapy alone for patients with T1G3 bladder cancer after TURBT,and there are very low rates of serious side effects.Intravenous chemotherapy may be considered as a new therapy strategy for T1G3 bladder cancer after TURBT.

Objective To investigate whether miR?485?3p plays a role in regulation of radiosensitivity of gastric cancer cells by targeting TLR1. Methods Quantitative real?time PCR and Western blot were used to determine the expression of miR?485?3p and TLR1, respectively. The interaction between miR?485?3p and TLR1 was verified by target prediction software ( DIANA, TargetScan, and miRanda) and dual luciferase reporter assay. Gastric cancer MGC803 cells transfected with miR?485?3p mimic or TLR1 siRNA were exposed to irradiation. Apoptosis assay, colony formation assay, and MTT assay were used to evaluate the changes in radiosensitivity of gastric cancer cells. Dual luciferase reporter assay was used to determine the effects of miR?485?3p overexpression and TLR1 silencing on the activity of NF?κB. Western blot was used to study the effects of miR?485?3p overexpression and TLR1 silencing on NF?κB target genes. Results In gastric cancer cells exposed to radiation, the expression of miR?485?3p was downregulated and the expression of TLR1 was upregulated. TLR1 was predicted to be the target of miR?485?3p by target prediction software. Dual luciferase reporter assay further confirmed TLR1 as the direct target of miR?485?3p. miR?485?3p negatively regulated the expression of TLR1. The overexpression of miR?485?3p, as well as TLR1 silencing, increased the apoptosis rate of cells, reduced colony formation and cell proliferation, and enhanced the radiosensitivity of the cells. Both miR?485?3p overexpression and TLR1 silencing reduced the activity of NF?κB and downregulated the expression of multiple NF?κB target genes. Conclusions miR?485?3p enhances the radiosensitivity of gastric cancer cells probably by targeting TLR1 and regulating the NF?κB signaling pathway.

OBJECTIVE:To investigate the efficacy and safety of Xiaojin pill combined with docetaxel,epirubicin and cyclo-phosphamide in the treatment of breast cancer in stage Ⅲ and its effect on immune functions. METHODS:102 patients with breast cancer in stage Ⅲ were randomly divided into control group (51 cases) and observation group(51 cases). All patients were given symptomatic treatments as hydration,alkalization and antiemetic treatments in the duration of chemotherapy,and were given conven-tional treatments as 0.75 mg Dexamethasome oral tablets in 12 and 6 hours before the chemotherapy lespectively. Control group was given 75 mg/m2 Docetaxel injection,d1,intravenous infusion+60 mg/m2 Epirubicin hyclrochloride for injection,d1,intravenous in-fusion+600 mg/m2 Cyclophosphamide for injection,d1,intravenous infusion;observation group was additionally given 6 g Xiaojin pill,orally,twice a day. 21 d was regarded as a treatment course. Short-term clinical efficacy,immunoglobulin,T lymphocyte sub-sets(CD4+/CD3+,CD8+/CD3+,CD4+/CD8+)and the incidence of adverse reactions before and after chemotherapy in 2 groups after 2 courses were observed. RESULTS:There were no significant differences in the short-term effective rate and incidence of adverse reactions between 2 groups (P>0.05). Before chemotherapy,there were no significant differences in the immunoglobulin and T lymphocyte subsets between 2 groups(P>0.05). After chemotherapy,immunoglobulin levels in 2 groups were significantly lower than before,the differences were statistically significant (P<0.05);but there were no significant differences between 2 groups (P>0.05);CD4+/CD3+,CD8+/CD3+,CD4+/CD8+ in control group and CD8+/CD3+ in observation group were significantly lower than before,and CD4+/CD3+ and CD4+/CD8+ in observation group were higher than control group,the differences were statistically significant (P<0.05),but there was no significant difference in the CD8+/CD3+ between 2 groups (P>0.05). CONCLUSIONS:Based on the conventional treatment,Xiaojin pill combined with docetaxel,epirubicin and cyclophosphamide shows significant effi-cacy in the treatment of breast cancer in stageⅢ,and can improve immune functions,with good safety.

PURPOSE: Allergic rhinitis (AR) is an inflammatory disorder of the upper airway. Exosomes or extracellular vesicles are nanosized vesicles of endosomal origin released from inflammatory and epithelial cells that have been implicated in allergic diseases. In this study, we characterized the microRNA (miRNA) content of exosomes in AR. METHODS: Extracellular vesicles were isolated from nasal mucus from healthy control subjects (n=10) and patients with severe AR (n=10). Vesicle RNA was analyzed by using a TaqMan microRNA assays Human Panel-Early Access kit (Applied Biosystems, Foster City, CA, USA) containing probes for 366 human miRNAs, and selected findings were validated with quantitative RT-PCR. Target prediction and pathway analysis for the differentially expressed miRNAs were performed using DIANA-mirPath. RESULTS: Twenty-one vesicle miRNAs were up-regulated and 14 miRNAs were under-regulated significantly (P<0.05) in nasal mucus from AR patients when compared to healthy controls. Bioinformatic analysis by DIANA-mirPath demonstrated that 32 KEGG biological processes were significantly enriched (P<0.05, FDR corrected) among differentially expressed vesicle miRNA signatures. Among them, the B-cell receptor signaling pathway (P=3.709E-09), the natural killer cell-mediated cytotoxicity (P=8.466E-05), the T-cell receptor signaling pathway (P=0.00075), the RIG-I-like receptor signaling pathway (P=0.00127), the Wnt signaling pathway (P=0.00130), endocytosis (P=0.00440), and salivary secretion (P=0.04660) were the most prominent pathways enriched in quantiles with differential vesicle miRNA patterns. Furthermore, miR-30-5p, miR-199b-3p, miR-874, miR-28-3p, miR-203, and miR-875-5p, involved in B-cell receptor and salivary secretion signaling pathways, were selected for validation using independent samples from 44 AR patients and 20 healthy controls. MiR-30-5p and miR-199b-3p were significantly increased in extracellular vesicles from nasal mucus when compared to healthy controls, while miR-874 and miR-28-3p were significantly down-regulated. In addition, miRNA-203 was significantly increased in AR patients, while miRNA-875-5p was found to be significantly decreased in AR patients. CONCLUSIONS: This study demonstrated that vesicle miRNA may be a regulator for the development of AR.
B-Lymphocytes ; Biological Processes ; Endocytosis ; Epithelial Cells ; Exosomes ; Humans ; MicroRNAs* ; Mucus* ; Receptors, Antigen, T-Cell ; Rhinitis* ; RNA ; Wnt Signaling Pathway

Objective To study the etiological agent of hand, foot and mouth disease ( HFMD) and the genetic characteristics of coxsackievirus A16 ( CVA16 ) strains isolated from clinical specimens of patients with HFMD in Liaocheng city in 2013.Methods Throat swab and stool specimens were collected from patients with HFMD in the disease surveillance hospitals in Liaocheng city from January to December 2013.Samples pos-itive for CVA16 strains were screened out for the isolation of virus strains with rhabdomyosarcoma ( RD) cells and Vero cells.The entire VP1 coding regions of 9 randomly selected CVA16 isolates were amplified and se-quenced.BioEdit and MEGA4 softwares were used for homology analysis.A phylogenetic tree among the 9 CVA16 isolates and 56 CVA16 representative strains of known genotypes and subgenotypes was constructed.Re-sults The results of PCR analysis showed that 747(77.73%) out of 961 specimens were positive for HFMD and among them, 74 samples (9.91%) were positive for EV71 strains, 130(17.40%) were CVA16 strains and 543(72.69%) were other enterovirus strains.The 9 CVA16 strains clustered into the B2b evolution branch of B genotype with the representative strains, sharing 97.7%to 100%homologies in nucleotide sequences and 99.3%to 100%in amino acid sequences.Conclusion Although EV71 and CVA16 strains were identified, other enteric viruses were the predominant pathogens causing HFMD in Liaocheng city in 2013.The CVA16 iso-lates belonged to B2b subgenotype.The pathogen spectrum of HFMD had already changed.It is necessary to strengthen the surveillance for EV71, CVA16 and other enteric viruses and understand their genetic characteriza-tions, which would be of great significance for the prevention and control of HFMD.