OBJECTIVETo explore the effects of microtubule depolymerization (MD) on the spontaneous beating rate, action potential (AP), and oxygen consumption of cardiac myocytes in rats and its mechanism.

METHODSOne-hundred and eighty neonatal SD rats divided into 12 batches were used in the experiment, and 15 rats in each batch were sacrificed for the isolation and culture of cardiac myocytes after the heart tissues were harvested. The cardiac myocytes were respectively inoculated in one 12-well plate filled with 6 round cover slips, one 12-well plate filled with 6 square cover slips, two cell culture flasks, and two cell culture dishes. After routine culture for three days, the cardiac myocytes from all the containers were divided into normal control group (NC, routinely cultured with 3 mL DMEM/F12 solution rewarmed at 37 °C for 3 h) and group MD (routinely cultured with 3 mL DMEM/F12 solution rewarmed at 37 ° and containing 8 µmol/L colchicine for 3 h) according to the random number table, with 3 holes, 1 flask, or 1 dish in each group. The morphological changes in microtubules were observed with confocal laser scanning microscope after immunofluorescent staining. The content of polymerized or dissociative α-tubulin was determined by Western blotting. Spontaneous beating rate of the cells was observed and calculated under inverted microscope. Dissolved oxygen concentration of DMEM/F12 solution containing cardiac myocytes was determined by oxygen microelectrode system before and after the addition of colchicine. Additionally, dissolved oxygen concentration of DMEM/F12 solution and colchicine + DMEM/F12 solution was determined. The whole-cell patch-clamp technique was used to record AP, delayed rectifier K+ current (I(K)), and L-type Ca2+ current (I(Ca-L)) in cardiac myocytes; current density-voltage (I-V) curves were drawn based on the traces. Data were processed with independent or paired samples t-test.

RESULTS(1) In group NC, microtubules of cardiac myocytes were around the nucleus in radial distribution with intact and clear linear tubiform structure. The microtubules in group MD were observed in dispersive distribution with damaged structure and rough linear tubiform structure. (2) In group MD, the content of dissociative α-tubulin of cells (0.61 ± 0.03) was obviously higher than that in group NC (0.46 ± 0.03, t = -6.99, P < 0.05), while the content of polymerized α-tubulin (0.57 ± 0.04) was significantly lower than that in group NC (0.88 ± 0.04, t = 9.09, P < 0.05). (3) Spontaneous beating rate of cells was (59 ± 8) times per min in group MD, which was distinctly higher than that in group NC [(41 ± 7) times per min, t = 5.62, P < 0.01]. (4) Dissolved oxygen concentration of DMEM/F12 solution containing cardiac myocytes was (138.4 ± 2.5) µmol/L, and it was reduced to (121.7 ± 3.6) µmol/L after the addition of colchicine ( t = 26.31, P < 0.05). There was no obvious difference in dissolved oxygen concentration between DMEM/F12 solution and colchicine + DMEM/F12 solution (t = 0.72, P > 0.05). (5) Compared with that of group NC, AP morphology of cells in group MD changed significantly, with unobvious repolarization plateau phase and shorter action potential duration (APD). The APD20, APD50, and APD90 were respectively (36.2 ± 3.8), (73.7 ± 5.7), and (115.1 ± 8.0) ms in group MD, which were significantly shorter than those of group NC [(40.2 ± 2.3), (121.4 ± 7.0), and (169.4 ± 5.6) ms, with t values respectively 2.61, 15.88, and 16.75, P values below 0.05]. (6) Compared with that of group NC, the I-V curve of I(K) of cells in group MD moved up with higher current density under each test voltage (0 to 40 mV) after activation ( with t values from 2. 70 to 3. 76, P values below 0.05) . (7) There was not much alteration in current density of I(Ca-L) under each test voltage (-30 to 50 mV) between 2 groups (with t values from -1.57 to 1.66, P values above 0.05), and their I-V curves were nearly overlapped.

CONCLUSIONSAfter MD, the I(K) is enhanced without obvious change in I(Ca-L), making AP repolarization faster and APD shortened. Then the rapid spontaneous beating rate increases oxygen consumption of cardiac myocytes of rats.

Action Potentials ; Animals ; Cells, Cultured ; Energy Metabolism ; Microtubules ; metabolism ; Mitochondria, Heart ; metabolism ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tubulin ; metabolism

Objective To investigate the value of fractional contrast medium bolus injection in improving the quality of CT portography.MethodsA total of 42 patients were randomly divided into two groups which were all given iohexol (350mgI/mL) as the contrast medium.20 patients in the group A were injected with conventional method (dosage of 100ml, rate of 4mL/s).The group B (22 patients) were treated with fractional contrast medium bolus injection, the first phase with 60mL contrast medium (rate of 4mL/s) and the second phase (10s delayed) with 40mL contrast medium (rate of 4m/s).The tube was washed by 20mL saline with the same rate of injection at both phases.The CT values and the image quality of the branches of the portal vein were evaluated according to the original and postprocessed images.Independent samples test was used to compare the CT values of the portal vein, splenic vein, superior mesenteric vein, hepatic parenchyma and portal vein-liver parenchyma.The subjective evaluation scores of the image quality were compared by wilcoxon.ResultsThe CT values of the portal vein, splenic vein and portal vein-liver parenchyma in the group B were significantly higher than that in the group A (t=3.317,3.523,P<0.01).There was no significant difference in CT values of hepatic parenchyma and superior mesenteric vein between the two groups.Subjective quality score in the group B was superior to that in the group A, and the difference was statistically significant.T The two evaluation physicians agreed well.ConclusionThe technique of fractional contrast medium bolus injection can significantly improve the image quality of CT portograghy.

Aim To investigate the role of endoplasmic reticulum stress in adenosine induced HepG2 cell apoptosis.Methods HepG2 cells were treated with different concentrations of ADO for 36 h,and the effect of ADO on cell proliferation was measured by MTT assay.Cell nuclei DAPI staining was used to detect the nuclei change after being treated with different concentrations ADO for 36 h or 2.0 mmol?L-1 ADO for different time.The effect of ADO on HepG2 cell cycle was analysed by flow cytometry after being treated with 2.0 mmol?L-1 ADO for 12 h or 24 h.Translocation of CHOP and Caspase-3 were measured by immunofluorescence after being treated with 2.0 mmol?L-1 ADO for 36 h.The proteins expressions of CHOP,Caspase-4,Caspase-3 and JNK were assayed by western blot.Results The viability of HepG2 cell decreased in a dose-dependent manner;the relative cell viability of 0.5,1,2,4,6 mmol?L-1 decreased by 13.48%?0.12%,27.92%?0.25%,35.21%?0.42%,51.46%?0.24%,71.42%?0.58%,compared with the control group respectively.The nuclei of HepG2 cells treated with different ADO concentrations or 2.0 mmol?L-1 ADO showed condensation,rounding and shrinkage,then fragmentation,which demonstrated cell apoptosis.After being treated with 2.0 mmol?L-1 ADO for 12 h or 24 h,cell cycle analysis showed sub-G1 phase increased;the apoptotic ratio of control,12 h and 24 h was 1.55%?0.12%,10.96%?0.07% and 21.04%?0.26% respectively.Immunofluorescence assay showed the CHOP and Caspase-3 translocation to the nuclei after being treated with 2.0 mmol?L-1 ADO.The expressions of CHOP,Caspase-3 and Caspase-4 increased after being treated with different concentrations ADO for 36 h,while the expression of JNK did not change.Conclusion Endoplasmic reticulum stress is involved in adenosine-induced HepG2 cell apoptosis.

Objective To investigate the effect of dexmedetomidine on perioperative cardiac function in patients with mild cardiac dysfunction undergoing laparoscopic radical resection of rectal cancer. Methods Sixty patients scheduled with laparoscopic radical resection of rectal cancer with mild heart failure were selected, with New York Heart Association (NYHA) cardiac function gradingⅠtoⅡ, American Association of Anesthesiologists (ASA) grading Ⅱ to Ⅲ, and age from 63 to 72 years. The patients were divided into dexmedetomidine group (group D) and control group (group C) according to the random digits table method with 30 cases each. At the beginning of induction, the patients in group D were given intravenous infusion a loading dose of dexmedetomidine at 0.5 μg/kg for more than 10 min. Then continuous intravenous infusion of dexmedetomidine was given at 0.3 μg/(kg·h) for 60 min. The patients in group C were given 0.9% sodium chloride with the same method. The small vessel resistance (SVR), left ventricular end-diastolic volume (LVEDV), left ventricular ejection fraction (LVEF), cardiac output (CO) and myocardial working index (Tei index) before induction (T0) and after administration of 20 min (T1), 40 min (T2), 60 min (T3) were measured by color Doppler ultrasound, and the heart rate (HR) and mean arterial pressure (MAP) were recorded at the same times. The time from the end of operation to extubation and incidences of agitation during recovery period were recorded. Results The T1to T3SVR in group D were significantly lower than those of T0: (883 ± 30), (827 ± 36) and (804 ± 38) dyn·s·cm-5vs. (1 075 ± 37) dyn·s·cm-5, and there were statistical differences (P＜0.05); compared with those in group C, the T1 to T3 SVR in group D were significantly lower, and there were statistical differences (P＜0.05). In group D, there were no statistical differences in CO between T1to T3 and T0(P＞0.05); compared with those in group D, the T1 to T3 CO in group C were significantly lower: (3.4 ± 0.6) L/min vs. (4.4 ± 1.0) L/min, (3.2 ± 0.7) L/min vs. (4.3 ± 0.8) L/min and (3.3 ± 0.9) L/min vs. (4.4 ± 0.9) L/min, and there were statistical differences (P＜0.05). In group D, there were no statistical differences in LVEF between T1to T3 and T0(P＞0.05); compared with those in group D, the T1to T2 LVEF in group C were significantly lower, and there were statistical differences (P＜0.05). In group D, there were no statistical differences in Tei index between T1 to T3 and T0(P＞0.05); compared with group D, the T1 to T2 Tei index in group C were significantly higher, and there were statistical differences (P＜0.05). There were no statistical differences in LVEDV after intra-group and inter-group comparison (P＞0.05). In group D, the T1 to T3 HR were significantly lower than T0: (68.1 ± 12.8), (67.3 ± 11.9) and (65.4 ± 11.7) times/min vs. (88.2 ± 15.1) times/min, and there were statistical differences (P＜0.05); compared with those in group C, the T1 to T3 HR in group were significantly slower. In group D, the T1 MAP significantly increased, significantly higher than those in T0 and in group C (P＜0.05). There was no statistical difference in the time from the end of operation to extubation between 2 groups (P＞0.05). The incidence of agitation during recovery period in group D was significantly lower than that in group C. Conclusions Dexmedetomidine can effectively promote the perioperative cardiac function recovery in patients with cardiac dysfunction undergoing laparoscopic radical resection of rectal cancer, suggesting that it has a certain myocardial protection effect.

Objective To investigate the correlation between salivary cystatin D level and salivary gland injury in patients with primary Sj?gren syndrome(pSS).Methods A total of 51 pSS patients admitted in the Department of Rheumatology and Immunology of the First Affiliated Hospital of Xi'an Jiaotong University from September 1,2022 to June 30,2023,and 51 age-and gender-matched healthy individuals who took physical examination in the hospital during same period were enrolled in the study.The level of salivary cystatin D was detected,and the difference in the level between the 2 groups was compared using an independent-samples t test.Pearson correlation analysis was applied to analyze the correlation between salivary cystatin D and clinical parameters in the patients.Results The pSS patients had significantly lower cystatin D level than the healthy controls(206.55±108.11 vs 374.32±172.24 pg/mL,P＜0.01).The cystatin D level in the pSS patients was positively correlated with both static(r=0.433,P=0.002)and dynamic salivary flow rates(r=0.363,P=0.009).The patients with higher score of salivary gland ultrasonography(SGUS)had obviously lower cystatin D than those with lower SGUS score(parotid gland:160.75±85.56 vs 290.53±95.17 pg/mL,P＜0.01;submandibular gland:157.76±87.59 vs 276.25±97.06 pg/mL,P＜0.01).The cystatin D level was also negatively correlated with peripheral blood IL-6 level(r=-0.453,P=0.001)and CD4+T cell count(r=-0.396,P=0.005)in the pSS patients.Conclusion Salivary cystatin D level can be used as an indicator of salivary gland damage for pSS patients.

Objective To evaluate the efficacy of Budesonide/formoterol to control asthma under real-life conditions.Methods A muhi-center, open label, non-interventional study was conducted.Asthma control after 12 week therapy with Budesonide/formoterol was assessed by Asthma Control Questionnaire (ACQ) and modified Asthma Control Questionnaire (ACQ5).Results A total of 360 asthma patients were recruited,including 228 adult patients and 132 child patients.After 12 weeks' therapy,all the patients' medium value of ACQ was decreased significantly from 2.03 (adults 2.20, children 1.74) at baseline to 0.60 (adults 0.78, children 0.29) (P < 0.0001), and the medium value of ACQ5 was also decreased significantly from 2.4 (adults 2.24, children 1.76) at baseline to 0.47 (adults 0.62, children 0.20) (P < 0.0001).Conclusion Budesonide/formoterol is effective in asthma treatment, by which most asthma patients obtain and maintain clineal control.

Objective: To analyze the clinicopathological features of the posttransplant lymphoproliferative disorders (PTLD) and to improve the diagnostic levels. Methods: The clinical data of 11 patients diagnosed with PTLD between January 2008 and January 2018 from Henan Provincial People's Hospital, Peking University Science Center and the Affiliated Third Hospital of Peking University were collected. The clinicopathological features and the potential prognostic predictors were retrospectively analyzed by using immunohistochemical staining, EB virus in situ hybridization, fluorescence in situ hybridization and gene sequencing. Results: There were 9 males and 2 females in 11 PTLD patients, and the median age of the total patients was 18 years old (3-34 years old). The median time of 9 cases who underwent hematopoietic stem cell transplantation developing PTLD was 4 months (2-24 months) after the transplantation. The other 2 cases undergoing solid organ transplantation (SOT) occurred PTLD after 6 months and 13 months, respectively. The lymph node was the most common site to be involved (9 cases), 1 case occurred in liver and 1 case occurred in nasopharynx site. Among 11 patients, 3 cases were classified as polymorphic PTLD (P-PTLD) and the other 8 cases were monomorphic PTLD (M-PTLD). EB virus of all cases was positive, and 8 cases of M-PTLD were classified as diffuse large B-cell lymphoma (DLBCL). Fluorescence in situ hybridization was used to detect bcl-2, myc, IGH and A20 gene, and only one case had the gene break of IGH, while other cases didn't find any other abnormalities. Ig gene clone analysis was made in 5 patients with PTLD, including 4 cases of M-PTLD with gene rearrangement and 1 case of P-PTLD without gene rearrangement. Univariate analysis showed that age (≤18 years old) was associated with poor prognosis (P = 0.040). Conclusions The clinicopathologic features of PTLD are various and infected by EB virus. Gene rearrangement can help the diagnosis.

Tooth germ injury can lead to abnormal tooth development and even tooth loss, affecting various aspects of the stomatognathic system including form, function, and appearance. However, the research about tooth germ injury model on cellular and molecule mechanism of tooth germ repair is still very limited. Therefore, it is of great importance for the prevention and treatment of tooth germ injury to study the important mechanism of tooth germ repair by a tooth germ injury model. Here, we constructed a Tg(dlx2b:Dendra2-NTR) transgenic line that labeled tooth germ specifically. Taking advantage of the NTR/Mtz system, the dlx2b⁺ tooth germ cells were depleted by Mtz effectively. The process of tooth germ repair was evaluated by antibody staining, in situ hybridization, EdU staining and alizarin red staining. The severely injured tooth germ was repaired in several days after Mtz treatment was stopped. In the early stage of tooth germ repair, the expression of phosphorylated 4E-BP1 was increased, indicating that mTORC1 is activated. Inhibition of mTORC1 signaling in vitro or knockdown of mTORC1 signaling in vivo could inhibit the repair of injured tooth germ. Normally, mouse incisors were repaired after damage, but inhibition/promotion of mTORC1 signaling inhibited/promoted this repair progress. Overall, we are the first to construct a stable and repeatable repair model of severe tooth germ injury, and our results reveal that mTORC1 signaling plays a crucial role during tooth germ repair, providing a potential target for clinical treatment of tooth germ injury.
Animals ; Mice ; Mechanistic Target of Rapamycin Complex 1/pharmacology* ; Signal Transduction ; Tooth/metabolism* ; Tooth Germ/metabolism* ; Odontogenesis

【Objective】 To investigate the clinical characteristics and risk factors of systemic lupus erythematosus （SLE） combined with symptomatic knee osteonecrosis （KON）. 【Methods】 We retrospectively analyzed the clinical data of 26 cases of SLE with KON treated in the Department of Rheumatology and Immunology， The First Affiliated Hospital of Xi’an Jiaotong University， from April 2013 to December 2019. 【Results】 The age of the 26 patients （2 males and 24 females） was （35.3±9.0） years old at the diagnosis of KON， and the course of SLE was （48.7±35.1） months. The time from glucocorticoids initiation to the development of KON was （37.8±42.7） months， the maximum dosage of methylprednisolone was （197.7±290.7）mg and the cumulative dosage was （6.02±6.66）g. Six of the patients had a history of large-dose glucocorticoids impulse therapy. All of them had a history of immunosuppressant treatment. SLEDAI score was （11.23±5.46） at the onset of SLE and （4.46±4.81） at the onset of KON. The most common initial symptoms were edema and arthritis. The most common systemic damages were blood system damage and lupus nephritis. The most common immunological abnormalities were positive antinuclear antibody （25/26）， positive anti-SSA/Ro52kD antibody （16/26）， and positive anti-SmD1 antibody （15/26）. There were 4 patients with positive anticardiolipin antibody （ACA）. Bone metabolism was characterized by vitamin D3 （Vit-D3） deficiency， insufficient N-terminal osteocalcin （N-OST）， and increased β-C-terminal telopeptide of type I collagen （β-CTx）. In 9 out of the 26 patients， SLE was combined with aseptic necrosis of the femoral head. Multifocal bone necrosis （at least 3 lesions） was common （12/26）. Longer disease course and glucocorticoids using time， larger cumulative dose and ACA positive were seen in patients with multifocal bone necrosis compared with those who had one lesion site. 【Conclusion】 KON most possibly occurs 3 to 4 years after the diagnosis of SLE， which is associated with high disease activity， large hormone dose， and long duration in the treatment process. Multifocal bone necrosis is easily seen in patients with severe disease， large initial hormone dose and high cumulative dose， as well as in ACA positive patients.

ABSTRACT OBJECTIVE To investigate the current situation of pediatric pharmacist-managed clinic in China, and to provide reference for pediatric pharmacist-managed clinic construction and improvement. METHODS Domestic Children's hospitals, Women's & Children's Hospital, and general hospitals with pediatric unit were selected as the survey objects, questionnaires were distributed through the Wenjuanxing Application, and SPSS 26.0 was used to describe the data. The development of pediatric pharmacist-managed clinic, the qualification of visiting pharmacists, the situation of post training and training needs were analyzed. RESULTS A total of 101 valid questionnaires were collected. Pediatric pharmacist-managed clinics had been set up in 55(54.5%) hospitals, of which 35 were independent pharmacists' clinics, and most had well medical document and patient management processes. However, about 70% of hospitals did not charge registration fees, and more than half of hospitals had fewer than 5 patient-visits per day. 85.5% of the hospitals were visited by clinical pharmacists, and about half of them were senior clinical pharmacists with more than 10 years of working experience. But only 3.6% of visiting pharmacists had the right of specific prescription. In 101 hospitals relevant post training for pharmacists had been carried out in 36 hospitals, of which 25 hospitals had set up pharmacist-managed clinic. In addition to pharmaceutical expertise, more than 50% of pharmacists had a strong demand for the improvement of physician-patient communication and problem-solving ability, and 30%-40% of the demand was focused on the collection of medication history, psychological counseling ability, and case-based learning in the pharmacist-managed clinic. CONCLUSION At present, the domestic pediatric pharmacist-managed clinic shows a vigorous development trend, however, there are insufficient registration fees, visits, and post training. Appropriate service methods of pediatric pharmacist-managed clinic should be actively explored, and a pediatric specialized training system for should be well constructed to improve the competency.