Objective:To investigate the relationship between mutated genes and clinical features in patients with essential thrombocythemia(ET).Methods:The clinical data of 69 patients with ET from October 2018 to March 2022 were retrospectively analyzed.According to driver mutation type,patients were divided into JAK2 group,CALR group and triple-negative group.The sex,age,cardiovascular risk factors,thrombosis,splenomegaly,routine blood test and coagulation status of patients in three groups were analyzed.Results:Among 69 ET patients,46 cases were associated with JAK2 mutation,14 cases with CALR mutation,8 cases with triple-negative mutation,and one with MPL gene mutation.There were no significant differences in age and sex among the three groups(P＞0.05).The highest thrombotic rate was 26.09％(12/46)in JAK2 group,then 12.5％(1/8)in triple-negative group,while no thrombotic events occurred in CALR group.The incidence of splenomegaly was the highest in JAK2 group(34.78％),while no splenomegaly occurred in triple-negative group.The white blood cell(WBC)count in JAK2 group was(9.00±4.86)× 109/L,which was significantly higher than(6.03±2.32)× 109/L in CALR group(P＜0.05).The hemoglobin(Hb)and hematocrit(HCT)in JAK2 group were(148.42±18.79)g/L and(0.44±0.06)％,respectively,which were both significantly higher than(131.00±15.17)g/L and(0.39±0.05)％in triple-negative group(P＜0.05).The platelet(PLT)in JAK2 group was(584.17±175.77)× 109/L,which was significantly lower than(703.07±225.60)× 109/L in CALR group(P＜0.05).The fibrinogen(Fg)in JAK2 and triple-negative group were(2.64±0.69)g/L and(3.05±0.77)g/L,respectively,which were both significantly higher than(2.24±0.47)g/L in CALR group(P＜0.05,P＜0.01).The activated partial thromboplastin time(APTT)in triple-negative group was(28.61±1.99)s,which was significantly decreased compared with(31.45±3.35)s in CALR group(P＜0.05).Conclusions:There are differences in blood cell count and coagulation status among ET patients with different driver gene mutations.Among ET patients,JAK2 mutation is most common.Compared with CALR group,the thrombotic rate,WBC and Fg significantly increase in JAK2 group,while PLT decrease.Compared with triple-negative group,the incidence of splenomegaly and HCT significantly increase.Compared with CALR group,Fg significantly increases but APTT decreases in triple-negative group.

Objective:To analyze the genes related to platelet activation in essential thrombocythemia (ET)based on transcriptome sequencing technology (RNA-seq ),and to explore the potential targets related to ET thrombosis. Methods:Blood samples from ET patients and healthy individuals were collected for RNA-seq,and differentially expressed lncRNAs,miRNAs,and mRNAs were selected to construct a lncRNA-miRNA-mRNA regulatory network. Differential mRNAs in the regulatory network were enriched and analyzed using Gene Ontology (GO ) and Kyoto Encyclopedia of Genes and Genomes (KEGG).The real-time PCR method was applied to validate differential mRNAs on crucial signaling pathways.Results:A total of 32 lncRNAs (3 up-regulated,29 down-regulated),16 miRNAs (8 up-regulated,8 down-regulated),and 35 mRNAs (27 up-regulated,8 down-regulated)were identified as differentially expressed.Among them,5 lncRNAs,12 miRNAs,and 19 mRNAs constituted the regulatory network.KEGG enrichment analysis showed that the differential mRNAs were related to the platelet activation signaling pathway,and there were 6 differential mRNAs related to platelet activation,namely F2R,ITGA2B,ITGB1,ITGB3,PTGS1,and GP1 BB,which were all up-regulated in their expression.RT-PCR results showed that the expression of five mRNAs including F2R,ITGA2B,ITGB1,ITGB3,and GP1BB were upregulated in ET patients compared with healthy subjects,and consistent with RNA-seq results,while PTGS1 expression was not significantly different.Conclusion:Differential mRNAs in ET patients are related to the platelet activation pathway,and F2R,ITGA2B,ITGB1,ITGB3,and GP1BB mRNAs may serve as novel targets associated with platelet activation in ET.

Objective To explore the clinical effect of hip arthroscopy in the treatment of borderline developmental dysplasia of the hip(BDDH)with acetabular labrum injury.Methods A total of 32 BDDH patients with acetabular labrum injury and undergoing hip arthroscopy between December 2019 and December 2021 were analyzed retrospectively.There were 8 males and 24 females,aged 39.41±16.27 years(18～59 years)and their mean lateral acetabular central-edge angle(LCEA)was 21.25±2.96°(18°～25°).Another 32 patients with acetabular femur impingement(FAI)matching in age and undergoing hip arthroscopy during the same period were selected into the control group,which includ-ed 15 males and 17 females,aged 42.78±12.18 years(24～70 years),with their average LCEA of 32.97°±4.96°(26°～42°).Both groups received the arthroscopic glenolabial repair,head and neck ar-ea plasty and joint capsule suture.Three,six and twenty-four hours after the surgery,both groups were evaluated the clinical effect using the modified Harris hip score(mHHS),hip outcome score sports specific subscale(HOS-SSS),hip outcome score-activities of daily living(HOS-ADL)and visual analogue scale(VAS).Results The BDDH and FAI groups were followed up for(27.6±5.4)and(28.3±6.5)months,respectively.There was no significant difference in all measurements between the two groups before the operation(P>0.05).However,3,12 and 24 months after surgery,the aver-age mHHS,HOS-SSS and HOS-ADL scores of both groups increased significantly,while the average VAS scores decreased significantly(P<0.001).Moreover,the average mHHS scores of BDDH group were significantly lower than FAI group 3,12 and 24 months after surgery(P<0.05).However,3 months after surgery,the average HOS-SSS and HOS-ADL scores in BDDH group were significantly lower than FAI group(P<0.05).None of the patients underwent re-surgery due to recurrence of symp-toms during the follow-up period.Conclusion Hip arthroscopy can achieve satisfying short-term out-comes in treating BDDH.

Objective:This study aims to establish an early warning diagnosis model for acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and to provide a simple, rapid, and accurate auxiliary diagnosis basis for clinical practice.Methods:The sample bank of subjects (patients admitted to the Eighth Medical Center of the PLA General Hospital from September 10, 2021, to July 25, 2023) was constructed, including the model establishment cohort [SCOPD group 49, 42 males and 7 females, (69.71±11.16) years old; AECOPD group 53, 49 males and 4 females, (72.60±10.19) years old] and the model validation cohort [SCOPD group 35, 28 males and 7 females, (69.97±10.40) years old; AECOPD group 35, 33 males and 2 females, (71.43±9.67) years old]. Fasting peripheral blood samples were collected, and the expression levels of IL-5, IL-17A, and IFN-α were detected by flow cytometry. Different expression levels were analyzed by Mann-Whitney U test. Binary logistic regression analysis was used to screen the related risk factors of COPD patients in acute exacerbation. The diagnostic efficacy of the model was evaluated by the receiver operating characteristic (ROC) curve.Results:The levels of IL-5 [1.64 (0.60, 2.86) pg/ml], IL-17A [1.42 (0.88, 2.29) pg/ml], and IFN-α [0.91 (0.59, 1.81) pg/ml] in the SCOPD group were significantly decreased compared with the AECOPD group IL-5 [4.68 (2.34, 9.40) pg/ml, Z=-5.033, P<0.001], IL-17A [2.33 (1.59, 4.62) pg/ml, Z=-3.919, P<0.001], IFN-α [2.83 (0.91, 3.75) pg/ml, Z=-4.127, P<0.01] in the cohort of model establishment. The results of binary logistic regression analysis between SCOPD and AECOPD groups showed that IL-5, IL-17A, and IFN-α were independent risk factors for acute exacerbation of patients with COPD ( P<0.05). And the regression equation is Y=-2.861+0.364×IL-5+0.385×IL-17A+0.445×IFN-α. The AUC value of IL-5, IL-17A, IFN-α and combined detection was 0.866 ( P<0.001). Compared to the SCOPD group and the AECOPD group in the cohort of model validation, the receiver operating characteristic (ROC) curve showed that the combined model of three (AUC=0.858, P<0.001) could be used to diagnose the AECOPD. And the Kappa value was 0.773( P<0.05). Conclusion:The combined detection of IL-5, IL-17A, and IFN-α has high diagnostic efficacy for patients with acute exacerbation of COPD. This method provides a new potential tool for the clinical diagnosis of AECOPD and has the value of further exploration and optimization, promotion, and application.

As a new clinical trial mode,decentralized & digitalized clinical trial(DCT)is based on digital health equipment and uses internet and artificial intelligence technologies to complete the screening,registration,randomization,intervention,evaluation and follow-up of subjects,which is helpful to improve efficiency and reduce trial costs.The DCT mode has been applied to evaluate the treatment and management effects of cardiovascular diseases such as atrial fibrilla-tion,heart failure,coronary heart disease,and hypertension,showing broad development prospects and application space.This article will provide a brief introduction to representative DCT in the global cardiovascular disease field,and look for-ward to the application prospects of this model,providing reference and guidance for accelerating the development of cardio-vascular DCT in China.

Objective To explore the in vitro and in vivo antimicrobial activity of pinaverium bromide(PVB)against Staphylococcus epidermidis(S.epidermidis).Methods S.epidermidis isolated from blood specimens of hospitalized patients in a hospital in Changsha from January to December 2022 were collected.Susceptibility test of S.epidermidis to PVB was performed using broth microdilution method and disc diffusion method.The time-and concentration-dependent antimicrobial activity of PVB were determined by time-killing assay.Ultrastructural chan-ges in bacteria after PVB treatment was observed by transmission electron microscope.The inhibitory and clearance effects of PVB on S.epidermidis biofilm were detected by crystal violet staining test.The combined antimicrobial effect of PVB and antimicrobial agents was studied through microdilution checkerboard technique.A skin abscess infection model was constructed to detect the in vivo antimicrobial activity of PVB.Results Antimicrobial suscepti-bility testing results showed that the minimum inhibitory concentration(MIC)and minimum bactericidal concentra-tion(MBC)of PVB against the standard strains RP62A and ATCC 12228 were 8 and 16 μg/mL,respectively;The MIC and MBC of clinical strains of S.epidennidis were 4-8 μg/mL and 8-16 μg/mL,respectively.Disc diffu-sion method results showed that compared with the untreated control group(0.60±0)cm,0.2 mg PVB treatment showed a significant inhibitory zone([2.26±0.09]cm;t=45.34,P＜0.001),and the diameter of inhibitory zone increased with the increase of PVB dosage.The time-killing curves indicated PVB had bactericidal activity,which enhanced with increased concentration and action duration.Transmission electron microscope observed that PVB could significantly damage the normal structure of S.epidermidis,leading to bacterial edema and lysis.In addition,at the concentration of 1 × MIC,PVB could significantly inhibit the formation of S.epidermidis biofilm,reducing the amount of biofilm formation(A570nm)from(2.30±0.18)to(0.47±0.11;t=14.85,P＜0.001).Meanwhile,PVB at the concentration of 1 × MIC could effectively destroy the formed biofilm,reducing the amount of biofilm from(2.64±0.10)to(1.77±0.30;t=4.76,P=0.009).The combination of PVB with amikacin and gentamicin exhibited synergistic antimicrobial activity,with synergistic inhibitory indexes of 0.50 and 0.31,respectively.Ani-mal models showed that 10 mg/kg body weight of PVB could reduce the area of abscesses from(68.83±10.68)mm2 to(35.50±10.58)mm2(t=6.52,P＜0.001),and reduce the amount of viable bacteria in abscesses from(6.11±0.55)lg(CFU/abscess)to(3.60±0.34)lg(CFU/abscess)(t=3.08,P=0.014).Hematoxylin-eosin staining revealed that the infiltration of inflammatory cells in skin abscesses in the PVB treatment group reduced significantly compared with the control group,tending to be normal.Conclusion PVB exhibits effective in vitro and in vivo an-timicrobial effect against S.epidermidis,which can be used as an alternative for the treatment of S.epidermidis-related infections.

Objective To perform laboratory detection and molecular traceability analysis on a case of imported kala-azar in Shenzhen to determine the infection strain. Methods Bone marrow puncture fluid and blood samples from a case of kala-azar in Shenzhen were collected for laboratory tests. The patient's bone marrow puncture fluid smears were stained with Giemsa and examined under a microscope. Blood samples were examined for antibodies using the rk39 visceral leishmania rapid diagnostic reagent. Whole blood DNA was extracted, and the ITS-1 sequence was amplified by PCR, sequenced and aligned, and a phylogenetic tree was constructed based on the ITS-1 sequence. Results Microscopic examination of the patient's bone marrow smears revealed a large number of Leishmania amastigotes without flagella, confirming the diagnosis of kala-azar. The patient's blood was tested positive with the rk39 rapid diagnostic reagent, and PCR amplification yielded an ITS-1 gene product sequence that matched the expected size. Sequence alignment with the NCBI database showed 100% sequence similarity with the ITS-1 gene sequence of Leishmania infantum, confirming the infecting strain as Leishmania infantum. Phylogenetic tree construction of the amplified ITS-1 sequence revealed clustering into a clade with Leishmania infantum , and close to KC347299, one of the reference sequence selected. Conclusions The case of kala-azar in Shenzhen was caused by Leishmania infantum. Kala-azar still occurs in China, so the diagnostic technology of medical personnel in non-epidemic areas should be strengthened so that they can actively use new diagnostic technologies to assist in diagnosis, thus improving their prevention and control ability of Leishmania parasites.

Objective To investigate the impact of propofol(Pro)on high glucose(HG)-induced myocardial cell injury through autophagy mediated by forkhead box O1(Fox O1)/thioredoxininteracting protein(TXNIP)signaling pathway.Methods H9c2 cells were divided into blank group(5.5 mmol·L-1 glucose),high glucose(HG)group(30 mmol·L-1 glucose),HG+Pro group(30 mmol·L-1 glucose+25 μmol·L-1 Pro),HG+Pro+negative control(OE NC)group(first transfected with OE NC,then treated with 30 mmol·L-1 glucose+25 μmol·L-1 Pro),HG+Pro+Fox O1 overexpression plasmid(Fox O1-OE)group(first transfected with Fox O1-OE,then treated with 30 mmol·L-1 glucose+25 μmol·L-1 Pro).Cell counting kit-8(CCK-8)method,TdT-mediated dUTP nick end labeling(TUNEL)method,enzyme-linked immunosorbent assay(ELISA),transmission electron microscope and Western blot were applied to detect the cell survival rate,apoptosis rate,superoxide dismutase(SOD)and malondialdehyde(MDA)levels in the supernatant,and the changes in Autophagosome,Fox O1/TXNIP and autophagy protein expression levels.Results The cell viabilities of blank group,HG group,HG+Pro group,HG+Pro+OE NC group and HG+Pro+Fox O1-OE group were(100.00±0.00)％,(48.15±4.82)％,(79.66±7.98)％,(78.89±7.91)％and(49.22±4.93)％,respectively;the apoptosis rates were(12.04±1.21)％,(42.34±4.25)％,(26.22±2.63)％,(27.02±2.71)％,(38.29±3.86)％,respectively;SOD levels were(62.24±6.25),(28.21±2.85),(55.37±5.58),(55.09±5.53),(30.66±3.08)U·mg-1 pro,respectively;MDA levels were(0.38±0.04),(1.43±0.15),(0.67±0.07),(0.72±0.08)and(1.34±0.14)U·mg-1 pro,respectively;the number of autophagosomes was 6.24±0.63,13.05±1.32,8.31±0.84,8.55±0.86 and 12.22±1.23,respectively;phosphorylated Fox O1(p-Fox O1)/Fox O1 ratios were expressed as 0.34±0.04,0.86±0.09,0.48±0.05,0.43±0.05 and 0.74±0.08;TXNIP were expressed as 0.24±0.03,0.62±0.08,0.38±0.04,0.36±0.04 and 0.58±0.06;Bcelin-1 were expressed as 1.12±0.12,0.53±0.06,1.02±0.11,1.05±0.11 and 0.62±0.07;p62 were expressed as 0.56±0.06,1.56±0.16,0.82±0.09,0.86±0.09 and 1.44±0.15;there were statistical differences in the above indexes between HG group and blank group,HG+Pro group and HG group,HG+Pro+Fox O1-OE group and HG+Pro+OE NC group(all P＜0.05).Conclusion Pro inhibits autophagy by inhibiting Fox O1/TXNIP signaling pathway,and improves HG-induced myocardial cell injury.

Orodispersible films (oral dispersible films), a novel form of oral solid dosage forms, are widely used for patients with dysphagia and those with uncontrollable autonomic behavior. In this study, suvorexant orodispersible film was prepared by hot melt extrusion technology, and the disintegration time, mechanical properties, in vitro dissolution and pharmacokinetics were evaluated, compared with other orodispersible films and commercially available tablets Belsomra. All experiments were approved by the Committee on the Management and Use of Laboratory Animals of Academy of Military Medical Sciences (IACUC-DWZX-2024-504). The results showed that the dissolution rate of suvorexant orodispersible film was faster and the mechanical strength was better than that of other commercially available orodispersible film, which could meet the needs of storage and transportation. Differential scanning calorimetry and X-ray diffraction results showed that suvorexant was dispersed within the orodispersible film in an amorphous state. The in vitro dissolution of the film was observed to be four times faster than that of the commercial tablets, achieving complete dissolution within five minutes. Pharmacokinetic evaluations in Beagle dogs revealed that the self-formulated orodispersible film exhibited no significant differences in the area under the blood concentration-time curve when compared with Belsomra. However, the film showed a faster onset of action, with a peak time that was twice as rapid, and a maximum blood concentration that was twice as high as that of Belsomra. Leveraging hot melt extrusion technology, the suvorexant orodispersible film offers a straightforward, continuous production process with consistent quality. It serves as an excellent platform for the development of solvent-free film preparations tailored for patients with special needs.

The rheological properties of drug and carrier materials have a wide range of guiding significance for the formulation and process development of solid dispersions. In this study, the rheological properties of materials with different drug carrier ratios were systematically studied with suvorexant as the model drug and copovidone as the carrier material, which provided a sufficient basis for determining the formulation and process of solid dispersions. The optimal suvorexant-copovidone ratio obtained by oscillating temperature scanning was 1∶4. If the ratio is greater than 1∶ 4, the glass transformation temperature of the material will increase significantly, and the solubilization effect of the solid dispersion will show a downward trend. The results of oscillation temperature scanning and oscillation temperature sweep can show that when the extrusion temperature is greater than 150 ℃, the viscosity of the material is less than 10 000 Pa·s, and the melt can be extruded smoothly, and the best extrusion temperature of 160-180 ℃ can be obtained by combining the dissolution results. Finally, the dissolution of suvorexant tablets guided by rheological property studies in multiple media is similar to that of the commercially available tablets Belsomra. Therefore, rheological studies can screen and optimize the formulation and process of suvorexant solid dispersions at the mechanism level, which is of great significance to improve the success rate of R&D and shorten the R&D cycle of solid dispersions prepared by hot melt extrusion.