Objective: To investigate the potential of conventional cranial computed tomography (CT) in assessing the early neurological deterioration(END), long-term prognosis, and traditional Chinese medicine (TCM) syndrome elements during the acute phase in patients with acute ischemic stroke (AIS). Methods: This study included 101 patients with AIS onset within 24 h in the Emergency Department of Fangshan Hospital, Beijing University of Chinese Medicine, from November 2019 to May 2021. To investigate the correlation between the relevant characteristics of the first conventional cranial CT in patients with AIS onset within 24 h and END, 90 d prognosis, and initial syndrome elements, the presence or absence of END, the 90 d prognosis (non-disabling outcome or functionally independent outcome), and the establishment of syndrome elements (internal fire, phlegm-dampness, blood stasis, qi deficiency, yin deficiency) were used as dependent variables and grouping criteria. Results: This study included 61 males and 40 females, with an age of (64.43±10.56) years. The time from onset to conventional cranial CT examination was 3.50 (1.50, 9.75) h. Among the patients, there were 70 cases (69.3%) of mild AIS, 30 cases (29.7%) of moderate AIS, and one case (1.0%) of severe AIS. Fifteen patients (14.9%) received intravenous thrombolysis. Among the 101 patients, six syndrome elements were observed within 24 h of onset: internal wind in 101 cases (100.0%), internal fire in 58 cases (57.4%), phlegm-dampness in 60 cases (59.4%), blood stasis in 67 cases (66.3%), qi deficiency in 39 cases (38.6%), and yin deficiency in 23 cases (22.8%). The incidence of END was higher in patients with lesions in the contralateral cerebral hemisphere to the affected limb (32.9%) than in those without such lesions (10.7%), showing a strong positive correlation with END occurrence (OR=4.082, P = 0.026). The incidence of END was higher in patients with lesions in the basal ganglia region (33.3%) and the carotid system blood supply area (32.8%) than in those without lesions in the basal ganglia region (15.8%) and the carotid system territory (14.7%), showing moderate positive correlations with END occurrence (OR=2.667, P =0.047; OR=2.836, P=0.044). The proportion of non-disabling outcomes was lower among patients with white matter degeneration (30.8%) and lesions in the contralateral cerebral hemisphere to the affected limb (52.1%) than in those without white matter degeneration (63.6%) and without such lesions in the contralateral cerebral hemisphere to the affected limb (78.6%), both showing strong negative correlations with the occurrence of non-disabling outcomes (OR=0.254, P=0.034; OR=0.296, P=0.015). Similarly, the proportion of functionally independent outcomes was lower among individuals with white matter degeneration (30.8%) and lesions in the contralateral cerebral hemisphere to the affected limb (64.4%) than in those without white matter degeneration (77.3%) and without such lesions in the contralateral cerebral hemisphere to the affected limb (89.3%), both also showing strong negative correlations with the occurrence of functionally independent outcomes (OR=0.131, P=0.001; OR=0.217, P=0.014). The incidence rates of internal fire, blood stasis, and yin deficiency syndrome elements were 66.7%, 73.0%, and 30.2%, respectively, among patients with lesions in the basal ganglia region, compared to 42.1%, 55.3%, and 10.5% among those without lesions in this region. The presence of lesions in the basal ganglia region showed moderate to strong positive correlations with internal fire and yin deficiency syndrome elements (OR=2.750, P=0.016; OR=3.670, P=0.028). Patients with lesions in the centrum semiovale and corona radiata regions (66.7%) had a higher incidence of qi deficiency than those without lesions in this region (33.7%), showing a strong positive correlation with the occurrence of qi deficiency (OR=3.931, P=0.022). No CT characteristics were found to be correlated with phlegm-dampness syndrome elements. Conclusion The first cranial CT in patients with AIS has potential application value in predicting disease progression, assessing prognosis, and diagnosing syndromes, which can provide physicians with diagnostic and treatment decisions to improve the long-term prognosis of patients with AIS.

Objectives: To elucidate the potential mechanisms of electroacupuncture (EA) in restoring detrusor-bladder neck dyssynergia (DBND) following suprasacral spinal cord injury (SSCI). Methods: A total of 52 specific pathogen-free (SPF) grade famale Sprague-Dawley (SD) rats (10 – 12 weeks, 250 – 280 g) were randomly assigned to either a sham group (n = 12) or a spinal cord injury model group (n = 40). In the model group, DBND was induced through Hassan Shaker spinal cord transection at T10 level, with 24 rats meeting inclusion criteria and subsequently randomized into DBND group (n = 12) and EA intervention group (DBND + EA group, n = 12). After spinal shock recovery (day 19 after modeling), DBND + EA group received EA treatment at Ciliao (BL32), Zhongji (RN3), and Sanyinjiao (SP6) acupoints for 20 min per session at 10/50 Hz frequencies, once daily for 10 d. Sham and DBND groups received anesthesia only without EA intervention. On day 29 post-modeling, all rats underwent urodynamic assessments, followed by hematoxylin and eosin (HE) staining, tandem mass tag (TMT) proteomics, and Western blot (WB) analysis of detrusor and bladder neck tissues. Differentially expressed proteins (DEPs) were defined as proteins with P < 0.05, unique peptides ≥ 2, and fold change > 1.2 or < 0.83. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was performed using KOBAS 3.0 (P < 0.01), and protein-protein interaction (PPI) networks were analyzed using Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) 11.5 and Cytoscape 3.9.1. Results: Compared with sham group, DBND group showed significantly elevated leak point pressure (LPP) and maximum cystometric capacity (MCC) (both P < 0.01). EA treatment significantly reduced both LPP and MCC compared with DBND group (P < 0.01 and P < 0.05, respectively). HE staining revealed that EA reduced detrusor fibrosis and improved bladder neck inflammation. TMT proteomics identified 30 overlapping DEPs in detrusor and 59 overlapping DEPs in bladder neck when comparing DBND + EA/DBND groups with sham group. In detrusor tissue, KEGG analysis revealed 10 significantly enriched pathways (P < 0.01), including mitogen-activated protein kinase (MAPK) signaling pathway. PPI analysis showed 22 of 30 DEPs were interconnected. In bladder neck tissue, 14 pathways were significantly enriched (P < 0.01), including relaxin signaling pathway, with 51 of 59 DEPs showing interconnections. Both TMT and WB validations demonstrated that compared with sham controls, DBND rats exhibited upregulated collagen type IV alpha 2 chain (Col4a2) and downregulated guanine nucleotide-binding protein G(z) subunit alpha (Gnaz) in detrusor tissue, while EA treatment normalized both proteins (both P < 0.05). In bladder neck tissue, DBND rats showed decreased expression of smoothelin (Smtn) and calcium-activated potassium channel subunit beta-1 (Kcnmb1) compared with sham controls (both P < 0.01), which were both upregulated following EA treatment (P < 0.01 and P < 0.05, respectively). Conclusion EA restores detrusor-bladder neck coordination in DBND through dual-target mechanisms. In detrusor tissue, EA modulates contraction via extracellular matrix remodeling, cyclic adenosine monophosphate (cAMP) signaling pathway regulation, and enhanced adenosine triphosphate (ATP) biosynthesis mediated by neurotransmitters. In bladder neck tissue, EA promotes relaxation by maintaining contractile phenotypes, reducing fibrosis, suppressing smooth muscle excitation, and regulating presynaptic neurotransmitter release. These findings provide mechanistic insights into EA's therapeutic role in managing DBND.

Objective:To analyze the mutation characteristics of rpoB gene in rifampicin-resistant Brucella strains. Methods:DNA of 4 rifampicin-resistant Brucella strains (JSY-26, G-9, WSY-13 and AW-3) isolated from Xinjiang Uygur Autonomous Region was selected, rifampicin rpoB gene was amplified by PCR and its nucleotide sequence was sequenced. The rpoB gene sequences of rifampicin-resistant Brucella standard strain (RB51) and sensitive strain (ALT-8) were used as reference, the mutation sites and types of the rpoB gene inside and outside the rifampicin resistance determination region (RRDR) of the 4 rifampicin-resistant Brucella strains were analyzed by Mega 7.0 software. Results:Through sequence alignment, both JSY-26 and WSY-13 strains underwent a single base point mutation at the RRDR 1 576 bp of the rpoB gene, with the base changing from guanine (G) to adenine (A). The G-9 strain underwent a single base point mutation at the RRDR 1 606 bp of the rpoB gene, with the base changing from cytosine (C) to A. The AW-3 strain showed 5 mutations of 3 types outside rpoB gene RRDR at 2 536, 2 537, 2 626, 2 636 and 2 654 bp, namely 3 insertion mutations [thymine (T) insertion once and C insertion twice], 1 deletion mutation (C deletion), and 1 single base point mutation (from G to C mutation).Conclusion:The RRDR mutations in the rpoB gene of the rifampicin-resistant Brucella strains are mainly characterized by single base point mutations, while multiple insertion and deletion mutations occur outside the RRDR.

ObjectiveTo explore the effects of Naozhenning granules on the memory function and neuron cells in the rat model of post-concussion syndrome based on mitochondrial biosynthesis. MethodSPF-grade Wistar rats were used to establish the multiple cerebral concussion （MCC） model by the weight-drop method. The successfully modeled rats were assigned into model， piracetam （0.324 g·kg^-1）， and low-， medium-， and high-dose （2.25， 4.5， and 9 g·kg^-1， respectively） Naozhenning groups. The rats were administrated with corresponding drugs by gavage and those in the blank group and model group were administrated the same volume of normal saline once a day for 14 days. The general state of rats was observed before and after treatment. The open field test and new object recognition test were conducted to examine the motor and memory abilities of rats. Hematoxylin-eosin staining was employed to observe the pathological changes of cortical neurons in rats. Western blot and real-time polymerase chain reaction were employed to determine the protein and mRNA levels， respectively， of peroxisome proliferator-activated receptor γ-coactivator-1α （PGC-1α）， nuclear respiratory factor-1 （NRF-1）， and transcription factor A mitochondrial （TFAM） in rat cortex. ResultCompared with the blank group， the model group showed anxious and manic mental status， yellow and messy fur， and reduced food intake. In the open field experiment， the model group showed reduced total movement distance， times of entering the central grid， and times of rearing decreased and increased resting time compared with the blank group （P<0.01）. The model group had lower recognition index of new objects than the blank group （P<0.01）. In addition， the modeling caused reduced neurons with sparse distribution and deformed， broken， and irregular nucleoli and down-regulated the mRNA and protein levels of PGC-1α， NRF-1， and TFAM in the cortex （P<0.01）. Compared with the model group， piracetam and Naozhenning improved the mental state， coat color， food intake， and activities of rats. In the open field test， piracetam and Naozhenning increased the total movement distance， the times of entering the central grid， and the times of rearing and shortened the resting time （P<0.05， P<0.01）. The piracetam and Naozhenning groups had higher recognition index of new objects than the model group （P<0.05， P<0.01）. Compared with the model group， the piracetam and Naozhenning groups showed increased neurons with tight arrangement and large and round nuclei， and some cells with irregular morphology and turbid cytoplasm. Furthermore， piracetam and medium-dose Naozhenning upregulated the protein levels of PGC-1α， NRF-1， and TFAM （P<0.01）. Low-dose Naozhenning upregulated the protein levels of NRF-1 and TFAM （P<0.01）， and high-dose Naozhenning upregulated the protein levels of PGC-1α and TFAM in the cortex （P<0.01）. The mRNA levels of PGC-1α， NRF-1， and TFAM in the cortex were upregulated in the piracetam group and Naozhenning groups （P<0.05， P<0.01）. ConclusionNaozhenning granules can improve the motor， memory， and learning， repair the neuronal damage， and protect the nerve function in the rat model of MCC by promoting mitochondrial biosynthesis.

Objective:To develop, validate and initially apply a joint function and health assessment scale for juvenile idiopathic arthritis patients.Methods:The first draft of the juvenile idiopathic arthritis joint function and health assessment scale was developed through literature analysis, discussion by the research team, semi-structured interviews, Delphi expert correspondence. From March to June 2024, a total of 260 children with juvenile idiopathic arthritis or their parents were prospectively recruited from Department of Rheumatology and Immunology, Children′s Hospital of Chongqing Medical University by convenience sampling method for pre-investigation and formal investigation.The reliability and validity of the scale were tested by item analysis, reliability analysis, exploratory factor analysis, content validity and criterion validity analysis, and the responsiveness of the scale to clinical changes was evaluated by estimating the minimum clinically important difference, and finally the formal scale was formed.Results:The juvenile idiopathic arthritis joint function and health assessment scale included disease activity assessment, daily activity and function assessment, pain, fatigue and disease outcome assessment, with a total of 5 dimensions and 24 items, in which the functional assessment subscale included 4 secondary dimensions and 18 items. The Cronbach′s α coefficient of the function assessment subscale was 0.88, the fold-half reliability was 0.86, and the test-retest reliability after 2-4 weeks was 0.84; the item-level content validity index was 0.80-1.00, and the scale-level content validity index was 0.93. Exploratory factor analysis extracted 4 common factors with a cumulative variance contribution of 70.0%. Preliminary application indicated the functional assessment subscale was moderately correlated with childhood health assessment questionnaire ( r=0.70, P<0.05), the total scale was strongly correlated with juvenile arthritis disease activity score-27 ( r=0.92, P<0.05), and moderately correlated with both active and limited joint count ( r=0.77, 0.68, both P<0.05). Reactivity analysis suggested that the minimum clinically important difference between the two visits of 41 children with clinical improvement and 25 children with disease activity was 0.49 (0.44, 0.54) and 0.51 (0.43, 0.58). Conclusion:The juvenile idiopathic arthritis joint function and health assessment scale has good reliability and validity, and has certain responsiveness to clinical changes, is simple and operable, and can be used as a tool for assessing joint function in children with juvenile idiopathic arthritis.

Objective:To construct A novel scoring model of immune signatures for colon cancer based on machine learning,which improve the survival prediction and immune therapy.Methods:Screening immune signatures from 1 301 immune-related genes(IRG)by the combined strategy of Lasso+bootstrap+multi Cox to calculate IRG scores of colon cancer patients from TCGA databases,and comprehensive the differences on function,prognostic status and immune therapy between high IRG scores group and IRG scores group.Results:Groups based on IRG scores were significantly different on the prognostic status of colon cancer patients,which were validated by other independent datasets.The IRG scores also could assess the effect of immune therapy of colon cancer.Conclusion:This study provides ideas for immune therapy and researches of colon cancer based on immune genes,and IRG scores can be used to assess the prognosis of colon cancer patients.

Objective:To evaluate the efficacy and safety of hypomethylating agents (HMA) in patients with myelodysplastic syndromes (MDS) .Methods:A total of 409 MDS patients from 45 hospitals in Zhejiang province who received at least four consecutive cycles of HMA monotherapy as initial therapy were enrolled to evaluate the efficacy and safety of HMA. Mann-Whitney U or Chi-square tests were used to compare the differences in the clinical data. Logistic regression and Cox regression were used to analyze the factors affecting efficacy and survival. Kaplan-Meier was used for survival analysis. Results:Patients received HMA treatment for a median of 6 cycles (range, 4-25 cycles) . The complete remission (CR) rate was 33.98% and the overall response rate (ORR) was 77.02%. Multivariate analysis revealed that complex karyotype ( P=0.02, OR=0.39, 95% CI 0.18-0.84) was an independent favorable factor for CR rate. TP53 mutation ( P=0.02, OR=0.22, 95% CI 0.06-0.77) was a predictive factor for a higher ORR. The median OS for the HMA-treated patients was 25.67 (95% CI 21.14-30.19) months. HMA response ( P=0.036, HR=0.47, 95% CI 0.23-0.95) was an independent favorable prognostic factor, whereas complex karyotype ( P=0.024, HR=2.14, 95% CI 1.10-4.15) , leukemia transformation ( P<0.001, HR=2.839, 95% CI 1.64-4.92) , and TP53 mutation ( P=0.012, HR=2.19, 95% CI 1.19-4.07) were independent adverse prognostic factors. There was no significant difference in efficacy and survival between the reduced and standard doses of HMA. The CR rate and ORR of MDS patients treated with decitabine and azacitidine were not significantly different. The median OS of patients treated with decitabine was longer compared with that of patients treated with azacitidine (29.53 months vs 20.17 months, P=0.007) . The incidence of bone marrow suppression and pneumonia in the decitabine group was higher compared with that in the azacitidine group. Conclusion:Continuous and regular use of appropriate doses of hypomethylating agents may benefit MDS patients to the greatest extent if it is tolerated.

Objective:To evaluate the effect of pancreatic duct stenting in the treatment of severe acute pancreatitis (SAP).Methods:The clinical data of 68 patients with SAP admitted to the Department of Hepatobiliary Surgery, General Hospital of Ningxia Medical University from January, 2019 to July, 2020 were retrospectively analyzed, including 38 males and 30 females, aged (44.85±8.51) years old. The patients were divided into two groups according to the treatment modality: the conservative group ( n=41) which received the conservative management, and the stent group ( n=27) which underwent pancreatic duct stent implantation and conservative management. The basic information, parenteral nutrition support time, fasting days, and other clinical data of patients were collected. Serum amylase, white blood cell count, acute physiology and chronic health (APACHE) II score and CT score were compared between the two groups at admission and 48 hours after treatment. Results:The serum amylase, white blood cell count, APACHE II score and CT score of the two groups were significantly lower after treatment (all P<0.05). Compared with the conservative group, the APACHE II score [(5.52±2.15) vs. (8.76±2.50)] and CT score [(4.85±1.96) vs. (6.18±1.07)] of patients were lower in the stent group after treatment (both P<0.05). Pancreatic duct stents were successfully implanted in all 27 patients of the stent group. Intubation to the main pancreatic duct failed in one patient, while the accessory pancreatic duct was alternatively intubated through the accessory nipple. No severe complications such as iatrogenic pancreatitis, bleeding, and perforation occurred after endoscopic retrograde cholangiopancreatography. The incidence of postoperative local complications was lower [18.52%(5/27) vs. 41.46%(17/41)], the proportion of antibiotic use>3 types [29.63%(8/27) vs. 56.10%(23/41)] decreased, and the analgesic episodes decreased [2(1, 3) vs. 4(3, 6)] in the stent group. The antibiotic usage duration [8(3, 11) d vs. 13(10, 19) d], the parenteral nutrition time [7(4, 15)d vs. 15(8, 18)d], the fasting time [5(3, 11) d vs. 11(6, 13)d] and the hospital stay [10(5, 16) d vs. 15(13, 23)d] were all shortened (all P<0.05). Conclusion:Both conservative management and pancreatic duct stenting can alleviate the clinical symptoms of SAP, and pancreatic duct stenting could help reduce local complications, relieve symptoms, and shorten hospital stay.

OBJECTIVES: This study aimed to compare the effects of virtual adjustment on occlusal interferences in mandibular posterior single crown and three-unit bridge restorations by using the mandibular movement track and the movement parameters of a virtual articulator. METHODS: Twenty-two participants were recruited. Digital casts of the maxillary and mandibular arches were obtained using an intraoral scanner, and the jaw registration system was used to record the data of the mandibular movement track and the movement parameters of the articulator. Four kinds of restorations with 0.3 mm occlusal interferences were designed with dental design software. In particular, single crowns were designed for teeth 44 and 46, whereas three-unit bridges were designed for teeth 44-46 and 45-47, and the corresponding natural teeth were virtually extracted. Virtual adjustment of the restorations was performed using two dynamic occlusal recordings, namely, the mandibular movement track and the movement parameters of the virtual articulator. A reverse-engineering software was used to measure the root-mean-square of the three-dimensional deviation of the occlusal surfaces between natural teeth and the adjusted restorations. The differences between the two methods of virtual-occlusion adjustment were compared and analyzed. RESULTS: For the same group of restorations, the three-dimensional deviation of the mandibular movement track group were lower than those of the virtual articulator group, and the differences were statistically significant (P<0.05). For the four groups of restorations adjusted by the same method, the three-dimensional deviation of the 46-tooth single crown was the largest and the smallest three-dimensional deviation was that of the 44-tooth single crown. Statistical differences existed between the 44-tooth single crown and the other groups (P<0.05). CONCLUSIONS For the occlusal design of posterior single crown and three-unit bridge, the mandibular movement track could be a more effective approach to virtual occlusal adjustment than the movement parameters of the virtual articulator.
Humans ; Mouth, Edentulous ; Occlusal Adjustment ; Jaw Relation Record ; Dental Articulators ; Software

The CRISPR/Cas systems comprising the clustered regularly interspaced short palindromic repeats (CRISPR) and its associated Cas protein is an acquired immune system unique to archaea or bacteria. Since its development as a gene editing tool, it has rapidly become a popular research direction in the field of synthetic biology due to its advantages of high efficiency, precision, and versatility. This technique has since revolutionized the research of many fields including life sciences, bioengineering technology, food science, and crop breeding. Currently, the single gene editing and regulation techniques based on CRISPR/Cas systems have been increasingly improved, but challenges still exist in the multiplex gene editing and regulation. This review focuses on the development and application of multiplex gene editing and regulation techniques based on the CRISPR/Cas systems, and summarizes the techniques for multiplex gene editing or regulation within a single cell or within a cell population. This includes the multiplex gene editing techniques developed based on the CRISPR/Cas systems with double-strand breaks; or with single-strand breaks; or with multiple gene regulation techniques, etc. These works have enriched the tools for the multiplex gene editing and regulation and contributed to the application of CRISPR/Cas systems in the multiple fields.
Gene Editing ; CRISPR-Cas Systems/genetics* ; Bacteria/genetics* ; Archaea ; Bioengineering