ObjectiveTo investigate the inhibitory effect of tauroursodeoxycholic acid （TUDCA） on neurogenic heterotopic ossification （NHO） after spinal cord injury and its potential mechanism. MethodsSPF-grade male C57BL/6 mice were used to establish an NHO model. The mice were randomly divided into three groups： sham operation group （laminectomy only）， NHO model group （spinal cord injury + intramuscular injection of cardiotoxin）， and TUDCA treatment group （NHO model + daily intragastric administration of TUDCA at 200 mg/kg for 4 consecutive weeks）. The area and maturity of heterotopic ossification were quantitatively assessed by micro-computed tomography（micro-CT） and hematoxylin-eosin（H&E） staining. Flow cytometry was used to quantify the interleukin（IL）-1β secretion level of M2 macrophages. RAW264.7 macrophages were cultured in vitro and divided into control， tumor necrosis factor（TNF）α， and TNFα+TUDCA （200 μmol/L） groups to detect the expression level of the polarization marker CD206. Additionally， macrophages were divided into control， TUDCA 200 μmol/L， and TUDCA 400 μmol/L groups to assess the IL-1β secretion level. ResultsCompared with the NHO group， the bone volume in the TUDCA-treated group was significantly reduced （12.69±1.13 vs. 7.647±1.19， P=0.003）. Compared with the TNFα group， qPCR results showed that the expression of the M2 macrophage marker CD206 was significantly upregulated in the TNFα+TUDCA group （0.55±0.019，1.11±0.023， P＜0.001）. Compared with the control group， ELISA results showed that TUDCA stimulation of macrophages significantly upregulated IL-1β secretion （404.7±24.38 vs. 652.8±32.61， P=0.003 7）. TUDCA administration upregulated IL-1β secretion by local muscle macrophages and inhibited heterotopic ossification formation. ConclusionsTUDCA might participate in inhibiting NHO formation by inducing M2 macrophage polarization and upregulating IL-1β secretion， thereby alleviating the development and progression of neurogenic heterotopic ossification after spinal cord injury. This finding provides experimental evidence for expanding the application of TUDCA in the treatment of complications following spinal cord injury.

Purpose To investigate the changes in quadriceps femoris microcirculatory perfusion level after low-load blood flow restriction training and its relationship with muscle strength.Materials and Methods Twenty-five healthy subjects were prospectively recruited in the First Affiliated Hospital of Kunming Medical University from September to November 2022.A 200 mmHg pressure cuff was applied at the root of the left thigh for blood flow restriction,and the subjects completed regular knee extension training within 4 weeks.Before the first training session and within 24 hours after the last training session,all subjects underwent scanning with the 3.0T MRI intravoxel incoherent motion sequence and the multi-echo steady-state acquisition three-dimentional imaging sequence.After image post-processing,the quadriceps femoris cross-sectional area,perfusion fraction and pseudo-diffusion coefficient were obtained,and the peak torque was measured using an isokinetic dynamometer.The MRI and muscle strength test parameters before and after training were compared,and correlation analyses were performed between the change of peak torque and the change of perfusion fraction,cross-sectional area,and pseudo-diffusion coefficient respectively.Results After low-load blood flow restriction training,the cross-sectional area of the left quadriceps femoris in subjects increased(t=-4.515,P<0.05).Among its components,the cross-sectional area of the left rectus femoris,vastus intermedius and vastus lateralis all increased(t=-3.302,-2.877,-3.207,all P<0.05).The perfusion fraction value of the left quadriceps femoris increased(t=-5.447,P<0.05);the perfusion fraction values of the left rectus femoris,vastus intermedius,vastus lateralis and vastus medialis all increased(t=-5.723,-4.621,-3.767,-4.682,all P<0.05);the muscle strength of the left quadriceps femoris increased(t=-3.983,P<0.05).There was a highly positive correlation between change of perfusion fraction and peak torque of the left quadriceps femoris in subjects(r=0.708,P<0.05).Conclusion After low-load blood flow restriction training,the changes in quadriceps femoris muscle microperfusion quantified by intravoxel incoherent motion are related to muscle strength,which is helpful for formulating rehabilitation training strategies for young patients.

Purpose To investigate the changes in quadriceps femoris microcirculatory perfusion level after low-load blood flow restriction training and its relationship with muscle strength.Materials and Methods Twenty-five healthy subjects were prospectively recruited in the First Affiliated Hospital of Kunming Medical University from September to November 2022.A 200 mmHg pressure cuff was applied at the root of the left thigh for blood flow restriction,and the subjects completed regular knee extension training within 4 weeks.Before the first training session and within 24 hours after the last training session,all subjects underwent scanning with the 3.0T MRI intravoxel incoherent motion sequence and the multi-echo steady-state acquisition three-dimentional imaging sequence.After image post-processing,the quadriceps femoris cross-sectional area,perfusion fraction and pseudo-diffusion coefficient were obtained,and the peak torque was measured using an isokinetic dynamometer.The MRI and muscle strength test parameters before and after training were compared,and correlation analyses were performed between the change of peak torque and the change of perfusion fraction,cross-sectional area,and pseudo-diffusion coefficient respectively.Results After low-load blood flow restriction training,the cross-sectional area of the left quadriceps femoris in subjects increased(t=-4.515,P<0.05).Among its components,the cross-sectional area of the left rectus femoris,vastus intermedius and vastus lateralis all increased(t=-3.302,-2.877,-3.207,all P<0.05).The perfusion fraction value of the left quadriceps femoris increased(t=-5.447,P<0.05);the perfusion fraction values of the left rectus femoris,vastus intermedius,vastus lateralis and vastus medialis all increased(t=-5.723,-4.621,-3.767,-4.682,all P<0.05);the muscle strength of the left quadriceps femoris increased(t=-3.983,P<0.05).There was a highly positive correlation between change of perfusion fraction and peak torque of the left quadriceps femoris in subjects(r=0.708,P<0.05).Conclusion After low-load blood flow restriction training,the changes in quadriceps femoris muscle microperfusion quantified by intravoxel incoherent motion are related to muscle strength,which is helpful for formulating rehabilitation training strategies for young patients.

Regenerating periodontal bone defect surrounding periodontal tissue is crucial for orthodontic or dental implant treatment. The declined osteogenic ability of periodontal ligament stem cells (PDLSCs) induced by inflammation stimulus contributes to reduced capacity to regenerate periodontal bone, which brings about a huge challenge for treating periodontitis. Here, inspired by the adhesive property of mussels, we have created adhesive and mineralized hydrogel microspheres loaded with traditional compound cordycepin (MMS-CY). MMS-CY could adhere to the surface of alveolar bone, then promote the migration capacity of PDLSCs and thus recruit them to inflammatory periodontal tissues. Furthermore, MMS-CY rescued the impaired osteogenesis and ligament-forming capacity of PDLSCs, which were suppressed by the inflammation stimulus. Moreover, MMS-CY also displayed the excellent inhibitory effect on the osteoclastic activity. Mechanistically, MMS-CY inhibited the premature senescence induced by the inflammation stimulus through the nuclear factor erythroid 2-related factor (NRF2) pathway and reducing the DNA injury. Utilizing in vivo rat periodontitis model, MMS-CY was demonstrated to enhance the periodontal bone regeneration by improving osteogenesis and inhibiting the osteoclastic activity. Altogether, our study indicated that the multi-pronged approach is promising to promote the periodontal bone regeneration in periodontitis condition by reducing the inflammation-induced stem cell senescence and maintaining bone homeostasis.
Animals ; Bone Regeneration/drug effects* ; Rats ; Periodontal Ligament/cytology* ; Microspheres ; NF-E2-Related Factor 2 ; Hydrogels ; Periodontitis/therapy* ; Osteogenesis/drug effects* ; Disease Models, Animal ; Stem Cells ; Male ; Rats, Sprague-Dawley ; Humans

Objective:To investigate the efficacy of Shuotong ureteroscope combined with flexible ureteroscope in the treatment of 2-3 cm lower calyceal calculi, and analyze the influencing factors.Methods:A total of 102 patients with lower calyceal calculi were treated in the Second People′s Hospital of Yulin from February 2019 to December 2022, and they were divided into the observation group and the control group, with 51 cases in each group. The patients of the observation group were treated with Shuotong ureteroscope combined with flexible ureteroscope, while the patients of the control group were treated with flexible ureteroscope. According to whether the stones were completely removed after operation, all patients were divided into non-stone removal group ( n=13) and stone removal group ( n=89). The operation time, hospitalization time, lithotripsy time, intraoperative blood loss, complication rate and stone clearance rate were compared between the observation group and the control group. Generalized Estimation Equation was used to analyze and evaluate the effects of treatment time, treatment scheme and their interaction on visual analogue scale (VAS), white blood cell (WBC), blood urea nitrogen (BUN), blood creatinine (Cr), hemoglobin (HGB) and procalcitonin (PCT). Univariate and multivariate Logistic regression were used to analyze the risk factors of stone removal rate. Nomogram model was constructed based on risk factors and evaluate the model. Results:Compared with the control group, operation time [(118.72±9.61) min vs (136.65±11.27) min], hospitalization stay [(6.43±1.12) d vs (10.29±2.23) d] and the lithotripsy time [ (51.23±10.38) min vs (56.62±11.43) min] of the observation group were shorter, and the amount of intraoperative blood loss [(128.52±10.20) mL vs (157.53±15.31) mL] were significantly less than those of the control group ( P< 0.05). The results of Generalized Estimation Equation analysis showed that treatment time, treatment regimen and their interaction had significant effects on WBC, HGB, BUN, Cr, PCT and VAS ( P< 0.05). Compared with the control group, the incidence of complications (5.88% vs 19.61%) of the observation group was lower and the stone clearance rate ( 94.12% vs 80.39%) was significantly higher than those in the control group ( P< 0.05). The mode of operation, infundibulopelvic angle(IPA), caliceal pelvic height (CPH) and the maximum diameter of stones were all influencing factors of stone removal rate in patients with 2-3 cm lower calyceal calculi. The nomogram model constructed in this study has good differentiation, calibration and clinical practicability, and can better identify high-risk patients with incomplete removal of 2-3 cm lower calyceal calculi. Conclusions:Shuotong ureteroscope combined with flexible ureteroscope is a safe, effective method for the treatment of 2-3 cm lower calyceal calculi. It has the advantages of simple operation, less intraoperative bleeding, less postoperative complications and high stone clearance rate. IPA, CPH, the maximum diameter of calculi and the mode of operation were all independent factors affecting the stone clearance rate of 2-3 cm lower calyceal calculi. The nomogram model constructed in this study can well identify the high-risk patients with incomplete clearance of 2-3 cm lower calyceal calculi.

Ultrafine powder and cell wall-broken powder of herbal medicine lack of the morphological characters and microscopic identification features. This makes it hard to identify herb's authenticity with traditional methods. We tested ITS2 sequence as DNA barcode in identification of herbal medicine in ultrafine powder and cell wall-broken powder in this study. We extracted genomic DNAs of 93 samples of 31 representative herbal medicines (28 species), which include whole plant, roots and bulbs, stems, leaves, flowers, fruits and seeds. The ITS2 sequences were amplified and sequenced bidirectionally. The ITS2 sequences were identified using Basic Local Alignment Search Tool (BLAST) method in the GenBank database and DNA barcoding system to identify the herbal medicine. The genetic distance was analyzed using the Kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic tree was constructed using MEGA 6.0. The results showed that DNA can be extracted successfully from 93 samples and high quality ITS2 sequences can be amplified. All 31 herbal medicines can get correct identification via BLAST method. The ITS2 sequences of raw material medicines, ultrafine powder and cell wall-broken powder have same sequence in 26 herbal medicines, while the ITS2 sequences in other 5 herbal medicines exhibited variation. The maximum intraspecific genetic-distances of each species were all less than the minimum interspecific genetic distances. ITS2 sequences of each species are all converged to their standard DNA barcodes using NJ method. Therefore, using ITS2 barcode can accurately and effectively distinguish ultrafine powder and cell wall-broken powder of herbal medicine. It provides a new molecular method to identify ultrafine powder and cell wall-broken powder of herbal medicine in the quality control and market supervision.

Objective To investigates factors affecting the positive rate of blocking antibody treated by paternal lymphocyte immunotherapy in patients with recurrent spontaneous abortion (RSA).Methods From January 2008 to August 2012,326 RSA cases undergoing treatment in Infertility Center of Qilu Hospital were studied retrospectively.Those patients were divided into 2 groups randomly:260 cases in intradermal injection group were administered via bilateral forearm intradermal injections for immunotherapy once 21 days,then the blocking antibody was determined after 2 (23 cases),3 (73 cases),4 (74 cases),5(90 cases) times respectively,while in subcutaneous injection group,the 66 cases were administered via subcutaneous injection once 21 days,the blocking antibody measured after 3 times; In both cases,the blocking antibody was all determined 2 weeks later.The positive rate of blocking antibodies and the rate of successful pregnancy was recorded,and then followed up after the blocking antibody turning positive.Results (1)Positive rate of blocking antibodies:the positive rate of blocking antibodies were 17％ (4/23),58％ (42/73),72％ (53/74) and 84％ (76/90) in the 2,3,4,and 5 times of intradermal injection group,respectively (P ＜ 0.05).In subcutaneous injection group,the positive rate of blocking antibodies was 38 ％ (25/66),which was significantly lower than that in group intradermal injection receiving 3 times immunotherapy (P ＜0.05).(2) The rate of pregnancy:the 176 patients out of 200 patients were pregnant when antibody was positive after immunotherapy,with 71.6％ (126/176)of patients gained successful pregnancy(the length of pregnancy more than 5 months).Conclusions The route and frequency of administration of immunotherapy could influence the positive rate of blocking antibody.The rate of successful pregnancy will be increased after blocking antibody turning positive.