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

A 82-year-old man was admitted to hospital with fever,unresponsiveness,elevated hypersensitive C-reactive protein and neutrophile granulocyte.Ceftriaxone was administrated by intravenous dripping in the emergency room,but the effect was not satisfactory.Following his admission to the ward,cefoperazone sulbactam were given.Elizabethkingia meningoseptica was identified by blood culture and further confirmed by 16S rRNA sequencing.The lumbar puncture showed that cerebrospinal fluid pressure was 80 mmH2O(1 mmH2O=0.0098 kPa)and biochemical results were normal.After 11 days of cefoperazone sulbactam treatment,the patient was discharged with negative blood culture.The hypersensitive C-reactive protein and neutrophile granulocyte had also declined.The patient received levofloxacin tablets for anti-infection treatment for 14 d after discharge.No signs of infection were observed in three months'following up.

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.

"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 access the therapeutic efficacy of newly-designed digital cholangioscope-assisted bedside one-stage lithotomy and biliary drainage for severe acute cholangitis caused by choledocholithiasis.Methods:Data of 26 patients were retrospectively analyzed, who were admitted into the intensive care unit (ICU), Zhongda Hospital, Southeast University, due to choledocholithiasis induced by severe acute cholangitis and underwent cholangioscope-assisted bedside one-stage lithotomy and biliary drainage from June 2020 to February 2022. Clinical outcomes were analyzed.Results:The time interval from disease onset to endoscopic intervention was 36.2±15.5 hours, with 7.2±4.9 hours from ICU admission to endoscopic intervention. Technical success rate was 100.0% in one-stage stone removal and biliary drainage. Except for one mild pancreatitis, no other complication occurred. Acute physiology and chronic health evaluation (APACHE)Ⅱ and sequential organ failure assessment (SOFA) scores prior to endoscopic intervention were 25.2±6.6 and 11.9±3.5, respectively. APACHE Ⅱ scores at day 1, 3, and 7 after endoscopic intervention were 21.7±6.5, 17.2±6.8 and 12.7±7.7, respectively, and SOFA scores were 10.6±2.9, 8.4±3.0 and 5.4±3.7, respectively, all of them were lower than those before operation ( P<0.001). The lengths of ICU stay and total hospitalization were 9.7±5.0 days and 12.8±4.5 days, respectively. In-hospital mortality occurred in 3 (11.5%) patients. According to a 6-month follow-up, one patient died of pneumonia, and another died of acute myocardial infarction. No acute cholangitis re-occurred in those survivors. Conclusion:Newly-designed digital cholangioscope-assisted bedside one-stage lithotomy and biliary drainage demonstrate significant improvements in prognosis, highlighting its safety in managing severe acute cholangitis.

Objective:To analyze the existing problems in maternal and child health services in Shanghai′s communities, so as to provide references for the continuous improvement and enhancement of maternal and child health work in community.Methods:Retrieve relevant literature on community maternal and child health services in Shanghai from databases such as CNKI, Wanfang, VIP, Chinese biomedical literature database and PubMed for text analysis, with a search period from 2009 to 2023. From August 2022 to May 2023, a purposive sampling method was used to select community health service centers in Shanghai for participatory observation. Interviews were conducted with maternal and child health service providers in community, management personnel at all levels, and obstetric experts from tertiary hospitals participating in the construction of medical alliances to understand the problems that exist in the process of providing maternal and child health services in community service centers. Based on text analysis, participatory observation, and interview results, a structure-process-outcome theory model was used to sort out a list of problems of maternal and child health services in Shanghai′s community. An electronic questionnaire was created based on the list of questions, and maternal and child health service personnel in each jurisdiction were invited to rate the existing problems from three aspects: importance, severity, and solvability. The key issues was determined based on the total score of each problem.Results:A list of 24 issues related to maternal and child health services in Shanghai′s community has been formed, with the structural dimension including team building and hardware facilities, the process dimension including mechanism construction, implementation process, and institutional coordination, and the result dimension only including system result category. According to the survey results, the three key questions with a total score greater than 7 were identified as the need for improvement in the maternal and child health knowledge and skills of service personnel; Insufficient staffing of service personnel, lack of dedicated personnel and specialized talents; The health information platform was not well-established, and institutions could not fully and timely share diagnosis and treatment data.Conclusions:This study has identified a problem list for maternal and child health care services in Shanghai′s communities, as well as key issues that urgently need to be addressed, which can provide reference for improving the quality of maternal and child health care services in Shanghai′s communities.

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.

Objective:To explore therapeutic efficacy and mechanism of puerarin(PUE)in treating of ulcerative colitis(UC).Methods:Network pharmacology and molecular docking technique were used to screen and analyze targets of PUE in regulating UC.C57BL/6 mice were given free access to 2.5%DSS aqueous solution for 7 days,and influence of PUE on changes in body weight and disease activity index(DAI)score were subsequently observed.Histopathological alterations of colon tissue were observed by HE staining,changes of goblet cell population in colon tissue were evaluated through Alcian blue staining;expressions of inflammatory factors in colon tissue were detected by qRT-PCR and ELISA.Effect of PUE on MODE-K cell viability and apoptosis were assessed by CCK-8 and flow cytometry.Results:A total of 38 common targets of PUE in modulating UC,such as AKT1,TNF,STAT3,CASP3,HIF1A and etc,mainly involving TNF,IL-17 and PI3K-Akt signaling pathway.In vivo experiments confirmed that PUE ameliorated degree of colon shortening,body weight and DAI scores and reduced inflammatory cell infiltration in mice.Besides,expressions of inflammatory factors in colon,such as TNF-α and IL-1β,were inhibited by PUE.Furthermore,in vitro experiments validated that PUE relieved DSS-induced apoptosis of epithelial cells.Conclusion:PUE alleviates occurrence and development of DSS-induced UC in mice.

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.