OBJECTIVE: To observe the efficacy and safety of Tiaowei Jiannao acupuncture (acupuncture for regulating defensive qi and nourishing brain) for post-ischemic stroke insomnia (PISI). METHODS: A total of 96 patients with PISI were randomized into an acupuncture group (32 cases, 1 case was excluded), a medication group (32 cases, 1 case dropped out, 1 case was excluded) and a sham-acupuncture group (32 cases, 1 case dropped out, 1 case was excluded). In the acupuncture group, Tiaowei Jiannao acupuncture was applied at bilateral Shenmai (BL62), Zhaohai (KI6), Hegu (LI4), Taichong (LR3), and Baihui (GV20), Sishencong (EX-HN1), Yintang (GV24⁺), Shenting (GV24), once a day, 1-day interval was taken after 6-day treatment, for 3 weeks totally. In the medication group, eszopiclone tablet was given orally, 1-3 mg a time, once a day for 3 weeks. In the sham-acupuncture group, non-invasive sham acupuncture was applied, the acupoint selection, frequency and course of treatment were the same as the acupuncture group. Before treatment, after 2,3 weeks of treatment, the scores of Pittsburgh sleep quality index (PSQI), self-rating sleep scale (SRSS), National Institutes of Health Stroke scale (NIHSS), Hamilton depression scale-17 (HAMD-17) were observed; before and after treatment, the sleep parameters were recorded using polysomnography (PSG); and the efficacy and safety were evaluated after treatment in the 3 groups. RESULTS: After 2,3 weeks of treatment, the scores of PSQI, HAMD-17 and SRSS in the acupuncture group and the medication group, as well as the SRSS scores in the sham-acupuncture group were decreased compared with those before treatment (P<0.05); after 2 weeks of treatment, the NIHSS score in the acupuncture group was decreased compared with that before treatment (P<0.05); after 3 weeks of treatment, the NIHSS scores in the acupuncture group, the medication group and the sham-acupuncture group were decreased compared with those before treatment (P<0.05). After 3 weeks of treatment, the scores of PSQI, SRSS, HAMD-17 and NIHSS in the acupuncture group and the medication group, as well as the NIHSS score in the sham-acupuncture group were decreased compared with those after 2 weeks of treatment (P<0.05). After 2,3 weeks of treatment, the scores of PSQI, SRSS and HAMD-17 in the acupuncture group and the medication group were lower than those in the sham-acupuncture group (P<0.05), the NIHSS scores in the acupuncture group were lower than those in the medication group and the sham-acupuncture group (P<0.05); after 3 weeks of treatment, HAMD-17 score in the acupuncture group was lower than that in the medication group (P<0.05), the NIHSS score in the medication group was lower than that in the sham-acupuncture group (P<0.05). Compared before treatment, after treatment, the total sleep time was prolonged (P<0.05), the wake after sleep onset, sleep latency, and non-rapid eye movement (NREM) sleep latency were shortened (P<0.05), the sleep efficiency was improved (P<0.05), the number of awakenings was reduced (P<0.05), the percentage of rapid eye movement (REM%) and the percentage of NREM stage 1 (N1%) were decreased (P<0.05), the percentage of NREM stage 2 (N2%) and the percentage of NREM stage 3 (N3%) were increased (P<0.05) in the acupuncture group and the medication group; the sleep latency was shortened in the sham-acupuncture group (P<0.05). After treatment, the PSG indexes in the acupuncture group and the medication group were superior to those in the sham-acupuncture group (P<0.05); in the acupuncture group, the number of awakenings was less than that in the medication group (P<0.05), the REM% and N1% were lower than those in the medication group (P<0.05), the N2% and N3% were higher than those in the medication group (P<0.05). The total effective rate were 93.5% (29/31) and 90.0% (27/30) in the acupuncture group and the medication group respectively, which were higher than 10.0% (3/30) in the sham-acupuncture group (P<0.05). There was no serious adverse events in any of the 3 groups. CONCLUSION Tiaowei Jiannao acupuncture improves the insomnia symptoms in patients with ischemic stroke, improves the quality of sleep, increases the deep sleep, promotes the recovery of neurological function, and relieves the depression. It is effective and safe for the treatment of PISI.
Humans ; Acupuncture Therapy ; Male ; Sleep Initiation and Maintenance Disorders/physiopathology* ; Female ; Middle Aged ; Aged ; Acupuncture Points ; Treatment Outcome ; Adult ; Ischemic Stroke/complications* ; Stroke/complications* ; Sleep

OBJECTIVES: The purinergic receptor P2X2 (P2RX2) encodes an ATP-gated ion channel permeable to Na⁺, K⁺, and especially Ca²⁺. Loss-of-function mutations in P2RX2 are known to cause autosomal dominant nonsyndromic deafness 41 (DFNA41), which manifests as high-frequency hearing loss, accelerated presbycusis, and increased susceptibility to noise-induced damage. Zebrafish, owing to their small size, rapid development, high fecundity, transparent embryos, and high gene conservation with humans, provide an ideal model for studying human diseases and developmental mechanisms. This study aims to generate a p2rx2 knockout zebrafish model using CRISPR/Cas9 gene editing system to investigate the effect of p2rx2 deficiency on the auditory system, providing a basis for understanding P2RX2-related hearing loss and developing gene therapy strategies. METHODS: Two CRISPR targets (sgRNA1 and sgRNA2) spaced 47 bp apart were designed within the zebrafish p2rx2 gene. Synthesized sgRNAs and Cas9 protein were microinjected into single-cell stage Tübingen (TU)-strain zebrafish embryos. PCR and gel electrophoresis verified editing efficiency at 36 hours post-fertilization (hpf). Surviving embryos were raised to adulthood (F0), tail-clipped, genotyped, and screened for positive mosaics. F1 heterozygotes were generated by outcrossing, and F2 homozygous mutants were obtained by intercrossing. Polymerase chain reaction (PCR) combined with sequencing verified mutation type and heritability. At 5 days post-fertilization (dpf), YO-PRO-1 staining was used to examine hair cell morphology and count in lateral line neuromasts and the otolith region. Auditory evoked potential (AEP) thresholds at 600, 800, 1 000, and 2 000 Hz were measured in nine 4-month-old wild type and mutant zebrafish per group. RESULTS: A stable p2rx2 knockout zebrafish line was successfully established. Sequencing revealed a 66 bp insertion at the first target site introducing a premature stop codon (TAA), leading to early termination of protein translation and loss of function. Embryos developed normally with no gross malformations. At 5 dpf, mutants exhibited significantly reduced hair cell density in the otolith region compared with wild type, although lateral line neuromasts were unaffected. AEP testing showed significantly elevated auditory thresholds at all 4 frequencies in homozygous mutants compared with wild type (all P<0.001), indicating reduced hearing sensitivity. CONCLUSIONS We successfully generated a p2rx2 loss-of-function zebrafish model using CRISPR/Cas9 technology. p2rx2 deficiency caused hair cell defects in the otolith region and increased auditory thresholds across frequencies, indicating its key role in maintaining zebrafish auditory hair cell function and hearing perception. The phenotype's restriction to the otolith region suggests tissue-specific roles of p2rx2 in sensory organs. This model provides a valuable tool for elucidating the molecular mechanisms of P2RX2-related hearing loss and for screening otoprotective drugs and developing gene therapies.
Animals ; Zebrafish/genetics* ; Receptors, Purinergic P2X2/deficiency* ; CRISPR-Cas Systems/genetics* ; Gene Knockout Techniques ; Phenotype ; Zebrafish Proteins/genetics* ; Disease Models, Animal

Objective:To investigate the peripheral blood levels of vitamin D and monocyte chemotactic protein 1 (MCP-1) in patients with Hashimoto thyroiditis (HT) and their clinical significance.Methods:Adopting a prospective research approach, 100 patients with HT from October 2022 to April 2023 in Jining First People′s Hospital were selected. Among them, the normal thyroid function was in 50 cases (HT normal thyroid function group), and the hypothyroidism was in 50 cases (HT hypothyroidism group). Another 50 cases of physical examination in the same period were selected as healthy control group. The general data were recorded. The levels of free triiodothyronine (FT 3), free thyroxine (FT 4), thyroid stimulating hormone (TSH), thyroglobulin antibody (TgAb), thyroid peroxidase antibody (TPOAb), vitamin D and MCP-1 were measured. Correlation analysis was performed using Pearson method or Spearman method. Binary Logistic regression analysis was used to analyze the risk factors leading to the development of HT, and the receiver operating characteristic (ROC) was used to assess the diagnostic value of vitamin D and MCP-1 for HT. Results:The FT 3 and FT 4 in HT hypothyroidism group were significantly lower than those in healthy control group and HT normal thyroid function group: (3.48 ± 1.00) pmol/L vs. (4.48 ± 0.49) and (4.28 ± 0.47) pmol/L, 12.40 (10.01, 14.23) pmol/L vs. 15.70 (14.30, 17.33) and 15.00 (13.10, 16.00) pmol/L, the TSH was significantly higher than that in healthy control group and HT normal thyroid function group: 8.60 (5.56, 27.13) mU/L vs. 1.97 (1.23, 2.89) and 3.06 (2.34, 3.42) mU/L, and there were statistical differences ( P<0.01); there were no statistical differences in FT 3, FT 4 and TSH between healthy control group and HT normal thyroid function group ( P>0.05). The TPOAb, TgAb and MCP-1 in HT normal thyroid function group and HT hypothyroidism group were significantly higher than those in healthy control group: 367.90 (151.60, 547.30) and 426.00 (175.30, 600.00) kU/L vs. 9.00 (9.00, 9.30) kU/L, 410.00 (222.00, 1 218.00) and 1 061.00 (427.30, 1 604.00) kU/L vs. 13.20 (12.08, 15.03) kU/L, 66.20 (54.43, 105.3) and 79.47 (41.57, 114.1) ng/L vs. 21.78 (15.23, 45.83) ng/L, the vitamin D was significantly lower than that in healthy control group: 14.32 (11.24, 16.99) and 12.73 (10.87, 15.36) μg/L vs. 18.12 (15.49, 21.92) μg/L, and there were statistical differences ( P<0.01); there were no statistical difference in TPOAb, TgAb, MCP-1 and vitamin D between HT normal thyroid function group and HT hypothyroidism group ( P>0.05). In healthy control group, there were no correlation between vitamin D, MCP-1 and FT 3, FT 4, TSH, TPOAb, TgAb ( P>0.05). In HT normal thyroid function group, vitamin D was positively correlated with FT 4 ( r = 0.376, P<0.01), negatively correlated with TPOAb ( r = - 0.400, P<0.01), and not correlated with FT 3, TSH and TgAb ( P>0.05); MCP-1 was positively correlated with TgAb ( r = 0.579, P<0.01), and not correlated with FT 3, FT 4, TSH, TPOAb ( P>0.05). In HT hypothyroidism group, vitamin D was positively correlated with FT 3 and FT 4 ( r = 0.522 and 0.567, P<0.01), negatively correlated with TSH, TPOAb and TgAb ( r = - 0.568, - 0.404 and - 0.328; P<0.01 or <0.05); MCP-1 was negatively correlated with FT 3 and FT 4 ( r = - 0.351 and - 0.469, P<0.05 or <0.01), positively correlated with TSH, TPOAb and TgAb ( r = 0.508, 0.431 and 0.522; P<0.01). In healthy control group, MCP-1 was not correlated with vitamin D ( P>0.05); in HT normal thyroid function group and HT hypothyroidism group, MCP-1 was negatively correlated with vitamin D ( r = - 0.457 and - 0.533, P<0.01). Binary Logistic regression analysis result showed that female, family history of thyroid disease, reduced vitamin D and elevated MCP-1were independent risk factors for the development of HT ( OR = 3.619, 3.675, 0.730 and 1.050; 95% CI 1.140 to 12.590, 1.174 to 16.220, 0.637 to 0.818 and 1.033 to 1.070; P<0.05 or <0.01). ROC curve analysis result showed that vitamin D, MCP-1 and vitamin D combined with MCP-1 had moderate diagnostic value for HT (area under the curve was 0.808, 0.858 and 0.886), and the combined diagnostic value was higher than that of the single index. Conclusions:In patients with HT, the serum vitamin D levels decrease and MCP-1 levels increase, which are related to thyroid function and antibodies. Insufficient vitamin D and increased MCP-1 can both increase the risk of HT.

Objective:To investigate the optimal serum antibody titer in acute stage for the diagnosis of Mycoplasma pneumoniae (MP) infection by passive agglutination, and to evaluate the clinical diagnostic value of different antibody titers.Methods:A cross-sectional study.Eighty-eight pairs of clinical serum samples were collected from children with MP infection treated at the Department of Pediatrics in Shengjing Hospital of China Medical University from December 2016 to February 2017 and Children′s Hospital of Baotou in November 2019.The four-fold change of the double serum specific antibody titer was used as the gold standard, and the receiver operating characteristic (ROC) curve was plotted.When detecting the single serum in acute stage, different antibody titers were used as positive criteria to evaluate their clinical application value in the diagnosis of MP infection and find the most appropriate serum antibody titer as the diagnostic cut-off value.Results:(1)When the serum specific antibody titer ≥1∶40 was used as the positive criterion, the sensitivity was 72.9%, the area under the ROC curve was 0.817, and the specificity was 87.5%, which might cause overdiagnosis.When the serum specific antibody titer ≥1∶160 was used as the positive criterion, the specificity was 97.5%, the area under the ROC curve was 0.775, and the sensitivity was 52.1%, which might cause missed diagnosis.When the serum specific antibody titer ≥1∶80 was used as the positive criterion, the sensitivity was 60.4%, the specificity was 97.5%, and the area under the ROC curve was 0.823, overall performing better compared with the said two criteria.(2)After the disease lasted at least 5 days, blood samples were collected.About 72.5% of the children had antibodies, and 60.0% of the children had antibody titers ≥1∶80.Conclusions:(1)When the passive agglutination method is used to detect MP infection, antibody titer ≥1∶80 is recommended as the diagnostic standard.However, in clinical practice, the diagnosis of MP infection depends on clinical and other laboratory test results.(2) It is appropriate to collect blood samples on 5-7 days of illness.If MP infection is clinically suspected, and an antibody titer of 1∶40 is also suggestive, it can perform cooperative diagnosis based on molecular biology lab results or retest at a shorter interval.

ObjectiveTo investigate the value of serum Fetuin-A and Fetuin-B combined with Homeostasis Model Assessment of Insulin Resistance （HOMA-IR） in predicting nonalcoholic fatty liver disease （NAFLD）. MethodsA total of 120 patients with NAFLD who attended Department of Gastroenterology， The First Affiliated Hospital of Dali University， from June 2020 to June 2021， and 120 healthy individuals who underwent physical examination at Physical Examination Center during the same period of time were enrolled as subjects， and clinical data were collected from all subjects. The serum levels of Fetuin-A and Fetuin-B were measured. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups， and the chi-square test was used for comparison of categorical data between two groups； the multivariate Logistic regression analysis was used to assess the risk factors for NAFLD. The receiver operating characteristic （ROC） curve was plotted to evaluate the predictive efficacy of Fetuin-A and Fetuin-B combined with HOMA-IR in NAFLD patients. ResultsCompared with the healthy control group， the NAFLD group had significantly higher levels of body mass index， systolic blood pressure， diastolic blood pressure， alanine aminotransferase， aspartate aminotransferase， fasting blood glucose， fasting insulin， triglycerides， HOMA-IR， Fetuin-A， and Fetuin-B （all P<0.05）. The multivariate Logistic regression analysis showed that Fetuin-A （odds ratio ［OR］=1.010， 95% confidence interval ［CI］： 1.001‍‍‍ ‍‍‍—‍‍‍ ‍‍1.020， P<0.05）， Fetuin-B （OR=1.113， 95%CI： 1.021‍ ‍‍—‍‍ ‍1.214， P<0.05）， and HOMA-IR （OR=24.053， 95%CI： 2.624‍ ‍‍—‍‍ ‍220.470， P<0.05） were independent risk factors for NAFLD. The ROC curve analysis showed that Fetuin-A， Fetuin-B or HOMA-IR alone had an area under the ROC curve （AUC） of 0.637 （95%CI： 0.551‍ ‍—‍ ‍0.722）， 0.853 （95%CI： 0.796‍ ‍—‍ ‍0.912）， and 0.837 （95%CI： 0.763‍ ‍—‍ ‍0.912）， respectively， and Fetuin-A combined with Fetuin-B， Fetuin-A combined with HOMA-IR， and Fetuin-B combined with HOMA-IR had an AUC of 0.853 （95%CI： 0.795‍ ‍—‍ ‍0.911）， 0.843 （95%CI： 0.770‍ ‍—‍ ‍0.916）， 0.922 （95%CI： 0.877‍ ‍—‍ ‍0.967）， respectively， while the combination of these three indicators had an AUC of 0.922 （95%CI： 0.877‍ ‍—‍ ‍0.966）. ConclusionFetuin-A and Fetuin-B have a certain value in predicting NAFLD， and Fetuin-B combined with HOMA-IR tends to have a higher predictive value.

OBJECTIVE To explore the effect of volatile oil of Ligusticum chuanxiong on the transdermal properties and cytotoxicity of triptolide in vitro. METHODS The chemical constituents of the volatile oil of L. chuanxiong were analyzed by gas chromatography-mass spectrometry. The lower abdominal skin of KM mice was separated and divided into triptolide group， triptolide in compatibility with volatile oil of L. chuanxiong groups at 1∶10， 1∶50， 1∶100 （hereinafter referred to as “compatibility 1∶10”“compatibility 1∶50”“compatibility 1∶100” groups）. After the skin of mice in each group was fully exposed to 0.2 g of the corresponding cream for 24 h， the cumulative transdermal dose （Qn） of triptolide in the receiving solution of each group was determined by high-performance liquid chromatography， and the transdermal absorption rate （Jss） was calculated. Human immortalized keratinocytes （HaCat） were used as a model， the CCK-8 method was used to detect the cell survival rate of different concentrations of the volatile oil of L. chuanxiong and triptolide before and after compatibility. RESULTS A total of 62 chemical constituents of the volatile oil of L. chuanxiong were identified， including Z-ligustilide， senkyunolide， and β-selinene. The Qn （P＜ 0.01） and Jss of triptolide increased within 24 h in the compatibility 1∶10 and 1∶50 groups， while the Qn （P＜0.05） and Jss decreased in the compatibility 1∶100 group as compared with the triptolide group. Compared with the triptolide group， the cell survival rate of HaCat was significantly increased in the compatibility 1∶10 and 1∶50 groups when the triptolide concentrations were 36， 72 and 144 ng/mL （P＜0.05 or P＜0.01）； while the cell survival rate of HaCat was decreased in the compatibility 1∶100 group， but the difference was not statistically significant （P＞0.05）. CONCLUSIONS When the compatibility ratio of triptolide and volatile oil of L. chuanxiong was 1∶10 or 1∶50， it can promote the transdermal absorption of triptolide and reduce the cytotoxicity of triptolide to HaCat.

"Forensic Science-Technical specifications for diatom inspection-Microwave digestion-vacuum filtration-microscopy"(GA/T 1662-2019)is currently the standard method used in forensic diatom testing.During the use of this method,the author found problems such as incomplete tissue digestion,excessive diatom fragments,too long suction filtration time,uneven baking of the filter membrane electric heating plate,and insufficient sample dryness.After continuous experimentation in the laboratory,some parameters of the inspection process were adjusted and the better expected results were achieved.The improved method only adds two basic equipment:a constant temperature water bath and a vacuum drying oven,making each experiment easy to realize.

Objective:To investigate alterations in the glymphatic system of spinocerebellar ataxia type 3 (SCA3) patients based on quantitative MRI, and its association with genetic information and motor dysfunction.Methods:The study was a cross-sectional study. This prospective study recruited 39 confirmed SCA3 patients (SCA3 group) and 40 matched healthy controls (HC group) who were seen at the Southwest Hospital of Army Medical University from May 2017 to June 2023. All subjects underwent cranial MRI scanning. Clinical assessments were conducted on all participants using the scale for the assessment and rating of ataxia (SARA) and the international cooperative ataxia rating scale (ICARS). The automatic segmentation and volume measurement of the choroid plexus based on Freesurfer 6.0; the perivascular interstitial space (PVS) was automatically segmented based on the deep-learning model VB-Net, and the volume of the PVS in each brain region was quantified after manual correction. Independent samples t-test and Mann-Whitney U-test were used to analyze the changes in the class lymphatic system in the SCA3 group and the HC group. Pearson partial correlation analysis was used to explore the relationship between CAG repeats, the glymphatic system, and motor dysfunction. Results:The standardized choroid plexus volume in the SCA3 group was (1.24±0.36)×10 3 mm 3, and that in the HC group was (0.96±0.34)×10 3 mm 3, with a statistically significant difference ( t=4.01, P<0.001). PVS volumes in the frontal lobe, temporal lobe, parietal lobe, basal ganglia, cerebellum, thalamus, and brainstem regions in the SCA3 group were significantly higher than those of HC group ( P<0.05). Partial correlation analysis revealed that CAG repeats in SCA3 group were positively correlated with SARA, ICARS, and basal ganglia PVS volumes ( r=0.65, 0.58, 0.29; P=0.001, 0.001, 0.042). Cerebellar and temporal lobe PVS volumes were positively correlated with SARA ( r=0.59, 0.47; P=0.001, 0.003), and positively correlated with ICARS scores ( r=0.61, 0.40; P=0.001, 0.011). Choroid plexus volume was positively correlated with cerebellar and basal ganglia PVS volumes ( r=0.41, 0.31; P=0.009, 0.043). Conclusions:The glymphatic system of SCA3 patients have significant alteration and have association with CAG repeats and motor dysfunction.

Hypophosphatemia rickets is a rare disease that is divided into two categories,namely,hereditary and ac-quirability.Its clinical manifestations include growth disorders,limb deformities and dysfunction,poor mineralization of the teeth,and growth retardation in children as well as hyperparathyroidism,osteoarthritis,osteomalacia,and pseudofrac-ture in adults.Oral manifestations include non-carious teeth with recurrent apical periodontitis,periapical abscess and even cellulitis,periodontitis,and early tooth loss.X-linked hypophosphatemia rickets(XLHR)accounts for approximate-ly 80％of all hypophosphatemia rickets.We report a 3-year-old child with XLHR whose first diagnosis was apical peri-odontitis of multiple non-carious and non-traumatic teeth.Through medical history,clinical examination,laboratory ex-amination,radiographic findings,genotype testing,and literature analysis,we analyze the pathogenesis,clinical manifes-tations,radiographic features,diagnosis and differential diagnosis,treatment,and follow-up.This work provides refer-ence for clinical diagnosis and treatment and reduces missed diagnosis and misdiagnosis by dentists.

Objective To explore the perioperative safety of video-assisted thoracic surgery (VATS) in Day Care Unit and the risk factors for delayed discharge under centralized management model. Methods The patients with VATS managed by the Day Care Unit of the Drum Tower Hospital Affiliated to Nanjing University Medical School in 2021 were retrospectively collected. The patients’ postoperative data and risk factors for delayed discharge were analyzed. Results A total of 383 patients were enrolled, including 179 males and 204 females with an average age of 46.09±14.82 years. Eleven (2.87%) patients developed grade 3-4 postoperative complications during the hospitalization. Eighteen (4.70%) patients visited unscheduled outpatient clinic within 7 days, and 6 (1.57%) patients were re-hospitalized within 30 days after discharge. The remaining patients had no significant adverse events during the 30-day follow-up. The average length of hospital stay was 2.27±0.35 d. The length of hospital stay was over 48 h in 48 (12.53%) patients. The independent risk factor for delayed discharge was lobectomy or combined resection (OR=3.015, 95%CI 1.174-7.745, P=0.022). Conclusion VATS can be safely conducted under the centralized management in Day Care Unit. The risk factor for delayed discharge is the extent of surgical resection.