Objective To investigate the effects of Echinococcus multilocularis infection on levels of memory T (Tm) cells and their subsets in lymph nodes of mice at different stages of infection, so as to provide new insights into immunotherapy for alveolarechinococcosis. MethodsTwenty-four C57BL/6J mice aged 6 to 9 weeks were randomly divided into the infection group and the control group, of 12 mice in each group. Mice in the infection group were administered with 3 000 E. multilocularis protoscoleces via portal venous injection, while animals in the control group were administered with an equal volume of physiological saline. Three mice from each group were sacrificed 4, 12 weeks and 24 weeks post-infection, and lymph nodes were sampled and stained with hematoxylin and eosin (HE) to investigate the histopathological changes of mouse lymph nodes in the infection group. The expression and localization of T lymphocyte surface markers CD3, CD4, and CD8 were observed in mouse lymph nodes using immunohistochemical staining. In addition, lymphocyte suspensions were prepared from mouse lymph nodes in both groups at different time points post-infection, and the levels of Tm cell subsets and their secreted cytokines were detected using flow cytometry. Results HE staining showed diffuse structural alterations in the subcapsular cortical and paracortical regions of mouse lymph nodes in the infection group 4 weeks post-infection with E. multilocularis. Immunohistochemical staining detected CD3, CD4 and CD8 expression in mouse lymph nodes in both groups. Flow cytometry revealed higher proportions of CD4⁺ Tm cells [(55.3 ± 4.8)% vs. (38.8 ± 6.1)%; t = -4.259, P < 0.05] and CD4⁺ tissue-resident Tm (Trm) cells [(57.7 ± 3.7)% vs. (34.1 ± 11.2)%; t = -3.990, P < 0.05] in mouse lymph nodes in the infection group than in the control group 4 weeks post-infection, and higher proportions of CD4⁺ Tm cells [(34.6 ± 3.2)% vs. (23.3 ± 7.5)%; t = -2.764, P < 0.05] and CD4⁺ Trm cells [(44.0 ± 1.9)% vs. (31.2 ± 1.5)%; t = -4.039, P < 0.05] in mouse lymph nodes in the infection group than in the control group 24 weeks post-infection. The proportions of CD8⁺ Tm cells were higher in the infection group than in the control group 4 weeks [(56.8 ± 2.7)% vs. (43.9 ± 5.2)%; t = -4.416, P < 0.01] and 12 weeks post-infection [(25.4 ± 2.7)% vs. (12.0 ± 2.6)%; t = -2.552, P < 0.05], while the proportions of tumor necrosis factor (TNF)-α⁺ CD4⁺ T cells [(15.7 ± 5.0)% vs. (49.4 ± 6.4)%; t = 7.150, P < 0.01], TNF-α⁺CD8⁺ T cells [(20.7 ± 5.5)% vs. (57.5 ± 8.4)%; t = -6.694, P < 0.01], and TNF-α⁺ CD8⁺ Tm cells [7.0% (1.0%) vs. 31.0% (11.0%); Z = -2.236, P < 0.05] were lower in the infection group than in the control group 24 weeks post-infection. Conclusions Tm cells levels are consistently increased in lymph nodes of mice at different stages of E. multilocularis infection, with Trm cells as the predominantly elevated subset. The impaired capacity of CD8⁺ Tm cells to secrete the effector molecule TNF-α in mouse lymph nodes at the late-stage infection may facilitate chronic parasitism of E. multilocularis.

OBJECTIVE To evaluate the palatability and chewability of chewable tablets， and provide reference for the quality evaluation of various types of chewable tablets. METHODS Using self-made Glucosamine hydrochloride chewable tablets as the model drug， the quality test was conducted. The in vitro simulation system for chewable tablets was established by using a texture analyzer and rheometer， and an oral simulation experiment was conducted on chewable tablets. The texture analyzer was used to measure the force required for chewing and simulate the static disintegration process of chewable tablets； the rheometer was adopted to measure the viscoelasticity， thixotropy， and deformability of chewable tablets during the chewing process. RESULTS The disintegration time limit， principal component content， and dissolution of self-made Glucosamine hydrochloride chewable tablets all met the limit requirements. The in vitro simulation results of the texture analyzer showed that self-made chewable tablets were easy to chew in both axial and radial directions， and the force required for chewing was within the range of the chewing force of the teeth； chewable tablets could disintegrate at an appropriate time without being chewed and only taken in the oral cavity. The in vitro simulation results of the rheometer showed that the chewable tablets in the oral cavity exhibited a behavior of elasticity as the main factor and viscosity as the secondary factor through the continuous stirring of the tongue， and the viscosity of the chewable tablets gradually decreased with tongue stirring or tooth chewing； when chewing with teeth， the internal force of the chewing tablets decreased， causing plastic deformation and crushing. After being crushed， the shape couldn’t be restored， making it easy to chew and swallow. CONCLUSIONS The combination of texture analyzer and rheometer can be used to simulate the oral chewing process and evaluate the palatability and chewability of self-made Glucosamine hydrochloride chewable tablets. This model can provide reference for the evaluation of various chewable tablets.

Objective To study the expression of thyroid hormone receptor binding protein 4(TRIP4)and DNA damage inducing transcription factor 4(DDIT4)in glioma tissue and their relationship with clinical pathological characteristics and prognosis.Methods 94 glioma patients admitted to the First Hospital of Hebei Medical University from February 2018 to February 2019 were selected as the research subjects.The expression of TRIP4,DDIT4 proteins in tissues were detected by immunohistochemistry.The relationship between the expression of TRIP4,DDIT4 proteins in glioma tissues and clinical pathological characteristics were compared.The differences in survival prognosis of glioma patients with different levels of TRIP4,DDIT4 protein expression were analyzed by Kaplan-Meier survival curve.Univariate and multivariate COX regression analysis was conducted to analyze the factors affecting the survival prognosis of glioma patients.Results The positive rates of TRIP4(68.09%),DDIT4(65.96%)proteins in glioma tissues were higher than those in adjacent tissues(13.83%,10.64%),with statistically significant differences(χ2=57.212,60.866,all P<0.05).There was a significant positive correlation between TRIP4 and DDIT4 protein expression in glioma tissues(r=0.722,P<0.05).The positive rates of TRIP4(83.64%vs 46.15%,80.00%vs 51.28%)and DDIT4(80.00%vs 46.15%,76.36%vs 51.28%)proteins in glioma tissues with tumor diameter≥3cm,WHO grade Ⅲ were significantly higher than those in tissues with tumor diameter＜3cm,WHO grade Ⅰ～Ⅱ(χ2=6.393～14.754,P<0.05).The 3-year overall survival rates of the TRIP4 positive and negative expression groups were 37.50%(24/64)and 66.67%(20/30),respectively.The 3-year cumulative survival of the TRIP4 positive expression group was significantly lower than that in the TRIP4 negative expression group(Log-rank χ2=5.949,P=0.015).The 3-year overall survival rate of DDIT4 positive and negative expression group was 37.10%(23/62)and 70.00%(21/30),respectively.The 3-year cumulative survival of the DDIT4 positive expression group was significantly lower than that in the DDIT4 negative expression group(Log-rank χ2=7.642,P=0.006).Tumor diameter≥3cm(HR=1.614,P=0.000),WHO grade Ⅲ(HR=1.790,P=0.000),positive TRIP4(HR=1.665,P=0.000)and positive DDIT4(HR=1.476,P=0.000)were independent risk factors affecting the survival prognosis of glioma patients.Conclusion The expression of TRIP4 and DDIT4 protein in glioma tissue was increased.Both of them were related to tumor diameter and WHO grade,and are potential tumor markers for survival prognosis of glioma.

To compare and analyze the "same disease, same price" policy in the regionsimplementing diagnosis related group(DRG) payment reform, and to provide recommendations for further policy optimization and extension.

Objective To explore the research hotspots and development trends of pyrroloquinoline quinone(PQQ),and explore the research and application value of PQQ.Methods In this study,Web of Science was used as the retrieval database to search for literature related to PQQ published from 1985 to 2022,and VOSviewer software was used to include the keywords,countries and regions,journals,etc.Bibliometric analysis was performed on these literatures.Results A total of 1 512 articles were included,with an overall upward trend in the number of annual publications.The journal category with the highest number of publications was Biochemistry and Molecular Biology.Journal of Biological Chemistry had the highest total link strength.The high-frequency keywords mainly included dietary nutrition supplement,mitochondria,antioxidant,etc.The visualization results of countries and regions showed that although China started its research relatively late,and it had a strong connection intensity with other countries.Conclusion In recent years,the research heat of PQQ has gradually increased,and its antioxidant effects,improvement of mitochondrial function and action targets may be the hotpots of future research.PQQ still has broad development prospects,and will inject new vitality into the pharmaceutical and healthcare industry.

Objective:To explore the genetic basis for a Chinese pedigree affected with Fragile X syndrome (FXS) through Nanopore long-read sequencing.Methods:A FXS pedigree who had undergone genetic counseling at the First Affiliated Hospital of Zhengzhou University in April 2023 was selected as the study subject. Nanopore long-read sequencing, triplet-repeat primed PCR (TP-PCR), methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) and trinucleotide polymorphism genotyping of androgen receptor (AR) gene were used to analyze the FMR1 CGG repeat number, methylation, and X chromosome inactivation of the pedigree members. This study has been approved by the Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University (No. KS-2018-KY-36). Results:Full mutation and CpG island hypermethylation were detected in the proband. The elder sister of the proband had full mutation of the FMR1 gene on one X chromosome and hypermethylation of CpG island, while the FMR1 gene on the other X chromosome was normal. FMR1 premutation was detected in the proband′s mother. Conclusion:Nanopore long-read sequencing can simultaneously detect the dynamic mutation and methylation status of the FMR1 gene on the two X chromosomes of females, which has important value for the diagnosis of FXS in different genders.

Objective To investigate the effect of bronchoscopic balloon dilation treatment of different intervals on the tracheal intubation-induced subglottic stenosis in children. Methods Children with tracheal intubation-induced subglottic cicatricial stenosis were selected as the research objects. Among the 36 children with pure balloon dilation treatment, 20 cases were enrolled in the observation group and 16 cases were enrolled in the control group. In the observation group, electronic bronchoscopy was performed at 3 days after the first-time balloon dilation treatment, and those with grade 2 or higher stenosis received the second-time balloon dilation treatment. The follow-up treatment principle was as follows. If there was obvious retraction compared with the previous time and the stenosis was grade 2 or higher, the balloon dilation treatment would be performed again after an interval of 3 days; if there was no obvious retraction, the evaluation would be performed again after an interval of 2 weeks until there was no obvious retraction in the stenosis site under endoscopy for 3 times in 6 consecutive weeks, and the degree of stenosis was less than 25%, and then the electronic bronchoscopy evaluation would be stopped. In the control group, electronic bronchoscopy was performed 7 days after the first-time balloon dilation treatment, and those with grade 2 or higher stenosis received the second-time balloon dilation treatment. The follow-up treatment principle was as follows. If there was obvious retraction compared with the previous time and the stenosis was grade 2 or higher, the balloon dilation treatment would be performed again after an interval of 7 days; the evaluation method for those without obvious retraction was the same as the observation group. The follow-up duration ranged from 6 to 9 months. Results There was no significant difference in the total number of treatments and outcomes between the two groups (P>0.05). The treatment duration of the observation group was significantly shorter than that of the control group (P < 0.05). There were no significant differences in intraoperative complications such as bleeding, oxygen saturation decrease, heart rate decrease, and blood pressure decrease during anesthesia between the two groups (P>0.05). There were no significant differences in postoperative fever, aggravated laryngeal stridor, and dyspnea between the two groups (P>0.05). There were no significant differences in dyspnea, activity tolerance, and the need for long-term oxygen therapy between the two groups (P>0.05). Conclusion Most of the tracheal intubation-induced subglottic cicatricial stenosis in children has a good medium- and long-term effect through pure balloon dilation treatment. Shortening the interval of pure balloon dilation to 3 days can shorten the total treatment duration, alleviate the psychological burden of parents, but does not increase their economic burden. There are no significant differences in intraoperative and postoperative complications, outcomes, and long-term follow-up compared with the original interval of 5 to 7 days.

Objective:To investigate the efficacy and safety of ab externo or ab interno transluminal trabeculotomy in the treatment of secondary glaucoma following pars plana vitrectomy (PPV).Methods:An observational case series method was performed.Seventeen eyes of 17 patients with glaucoma following PPV were enrolled in Henan Eye Hospital and Beijing Tongren Hospital from May 2016 to Feburary 2022.Primary conditions of patients receiving PPV included retinal detachment in 13 eyes, vitreous hemorrhage in 3 eyes, and entophthalmia in 1 eye.All the subjects underwent ab externo (11 eyes) or ab interno (6 eyes) transluminal trabeculotomy.The scope of all cases accepted trabeculotomy was ≥300°(11 cases of 360°, 4 cases of 330° and 2 cases of 300°).Before and at 1 week, 1 month, 6 months and 12 months after surgery, the intraocular pressure (IOP) was evaluated by Goldmann Tonometer and the best corrected visual acuity (BCVA) was measured using a standard visual acuity chart and converted to logrithm of minimal angle of resolution (LogMAR).The number of anti-glaucoma drug applications and surgery-related complications were recorded.The primary outcomes evaluated were IOP and surgical success rate.Secondary outcomes were medication quantity, BCVA (LogMAR) changes, and complications.Surgical success was defined as IOP reduction to <21 mmHg (1 mmHg=0.133 kPa) with or without the use of IOP-lowering medication.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2021[41]).Written informed consent was obtained from each subject.Results:Preoperative, 1-week, 1-month, 6-month, and 12-month postoperative mean IOP was (34.41±5.11), (21.88±11.72), (20.77±7.67), (19.50±7.01), and (16.32±4.68)mmHg, respectively, with an statistically significant overall difference ( F=20.779, P<0.001).IOP at difference time points after surgery were lower than that before surgery, showing statistically significant differences (all at P<0.01).Compared with before surgery, IOP was reduced more than 40% at 12 months after surgery in 14 eyes.Surgical success rates at 6 and 12 months after surgery were both 76.5%.The number of IOP-lowing drugs decreased significantly after operation ( Z=-4.580, P<0.001).The difference in BCVA between before and 6 months after surgery was not statistically significant ( Z=-1.311, P=0.190).No serious complications were seen in any of the operated eyes postoperatively. Conclusions:Ab externo or ab interno transluminal trabeculotomy is safe and effective in the treatment of secondary glaucoma after PPV.

OBJECTIVE: To explore the genetic etiology for a Chinese pedigree affected with Meckel syndrome. METHODS: A pedigree with a history of three consecutive adverse pregnancies which presented at the First Affiliated Hospital of Zhengzhou University on August 31, 2017 was selected as the study subject. Clinical data of the pedigree were collected. High-throughput sequencing was carried out to screen for variants of ciliopathy-related genes in the third fetus following induced abortion, and candidate variant was verified by Sanger sequencing. RESULTS: The first pregnancy of the couple had ended as spontaneous abortion, whilst the fetus of the second pregnancy was suspected for having ciliopathy, though no genetic testing was carried out following elected abortion. The fetus of the third pregnancy was suspected for having ciliopathy, and high-throughput sequencing and Sanger sequencing had shown that the fetus had harbored compound heterozygous variants of the TMEM67 gene, including c.978+1G>A from the father and c.1288G>C (p.D430H) from the mother. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.978+1G>A was classified as a pathogenic variant (PVS1+PM2_Supporting+PP5), whilst the newly discovered c.1288G>C (p.D430H) was classified as a likely pathogenic variant (PM2_Supporting+PM3+PM5+PP3). CONCLUSION The c.978+1G>A and c.1288G>C (p.D430H) compound heterozygous variants of the TMEM67 gene probably underlay the three consecutive adverse pregnancies suspected for ciliopathy in this pedigree. The discovery of c.1288G>C (p.D430H) has also expanded the mutational spectrum of the TMEM67 gene.
Female ; Pregnancy ; Humans ; Pedigree ; East Asian People ; Ciliary Motility Disorders/genetics* ; Ciliopathies ; Abortion, Spontaneous ; Membrane Proteins/genetics*

Objective:To investigate the predictive value of the changes of platelet-lymphocyte ratio (PLR) and neutrophil-lymphocyte ratio (NLR) in the therapeutic efficacy of the neoadjuvant treatment for breast cancer.Methods:The clinical data of 72 breast cancer patients who received neoadjuvant therapy in Nantong Tumor Hospital between February 2020 and January 2022 were retrospectively analyzed and the changes of PLR and NLR before and after neoadjuvant therapy were also analyzed. The receiver operating characteristic (ROC) curves were used to assess the predictive value of PLR, NLR and their changes in pathological complete remission (pCR) after neoadjuvant therapy.Results:The area under the ROC curve of PLR and NLR before the treatment, the difference in PLR before and after the treatment (ΔPLR), the difference in NLR before and after the treatment (ΔNLR) in predicting pCR was 0.520, 0.505,0.724 and 0.686,and the corresponding cut-off value was 269.231, 2.559, -2.840 and -1.457; the patients were divided into high and low groups according to the cut-off values. NLR before the treatment was not correlated with clinicopathological characteristics (all P > 0.05),while PLR before the treatment was correlated with tumor size ( P = 0.029), and ΔPLR was correlated with progesterone receptor expression ( P = 0.025), human epidermal growth factor receptor 2 (HER2) expression ( P < 0.001), molecular subtype ( P < 0.001), N stage ( P = 0.002), clinical stage ( P = 0.002) and treatment modality ( P < 0.001). ΔNLR was associated with HER2 expression ( P = 0.002), molecular subtype ( P = 0.024), tumor size ( P = 0.007), neural invasion ( P = 0.006), N stage ( P = 0.006), clinical stage ( P = 0.016) and treatment modality ( P = 0.014). ΔPLR and ΔNLR were influencing factors for patients achieving pCR after neoadjuvant therapy (all P < 0.05). Conclusions:Stage Ⅲ invasive breast cancer patients with higher ΔPLR and ΔNLR after neoadjuvant therapy have better prognosis.