OBJECTIVE To analyze the differences in chemical components of Gardenia jasminoides before and after processing with ginger， and to evaluate the quality differences among different producing areas. METHODS Ultra-high performance liquid chromatography-tandem time-of-flight mass spectrometry was used to analyze the compositional differences of G. jasminoides before and after processing with ginger. The water content， total ash， and ethanol-soluble extract content of ginger- processed G. jasminoides were determined according to the 2020 edition of Chinese Pharmacopoeia. High performance liquid chromatography was adopted to determine the contents of genipin gentiobioside， geniposide， crocin Ⅰ and crocin Ⅱ in ginger- processed G. jasminoides. RESULTS A total of 49 chemical components were identified from raw G. jasminoides and ginger- processed G. jasminoides， including 14 flavonoids， 15 iridoids， 10 organic acids， 2 alkaloids and 8 other compounds. Among them， 42 components were detected in raw G. jasminoides， 28 in ginger-processed G. jasminoides， and 21 components were common to both. After processing with ginger， raw G. jasminoides lost 21 components （including iridoids， flavonoids， alkaloids， and others）， while 7 chemical components were added （including coumarins， organic acids， organic acid esters， and flavonoids）. For the 15 batches of ginger-processed G. jasminoides， the water content ranged from 5.64% to 7.11%， total ash from 2.92% to 4.87%， and ethanol-soluble extract from 40.61% to 58.02%. The average contents of genipin gentiobioside， geniposide， crocin Ⅰ and crocin Ⅱ were 0.108 7， 0.542 2， 0.565 0， and 0.012 5 mg/g， respectively. CONCLUSIONS After processing with ginger， G. jasminoides loses 21 components， while 7 new components are added. Differences are observed in the water content， total ash， ethanol-soluble extract， and the contents of genipin gentiobioside， geniposide， crocin Ⅰ， and crocin Ⅱ of ginger-processed G. jasminoides from different producing areas. Notably， samples from Fujian exhibit high contents of genipin gentiobioside and ethanol-soluble extract， while samples from Jiangxi have a high content of crocin Ⅰ.

Objective:To analyze the differences in emergency rescue service capacities between county-level hospitals in Zhejiang and Jiangsu provinces, and propose targeted improvement suggestions.Methods:Cluster sampling was employed, with representative county-level hospitals recommended by provincial hospital alliances in Zhejiang and Jiangsu. Emergency departments voluntarily submitted data, resulting in 24 hospitals from Zhejiang and 34 from Jiangsu. Data were collected through the China County-Level Hospital Emergency Service Capability Evaluation System. Differences between provinces were analyzed. Spearman correlation analysis explored relationships between multidimensional indicators, and multivariate logistic regression identified factors influencing hospital grade B++ and above.Results:In terms of department scale, Jiangsu had a larger emergency department area (4 853 m 2vs. 1 927 m 2, P<0.001), more beds of comprehensive inpatient wards in the emergency department (25.5 beds vs. 5.5 beds, P=0.011), and higher scores for the ratio of emergency department area to beds (4.8 vs. 3.4, P=0.005) than Zhejiang. In terms of staffing, Jiangsu had a higher proportion of doctors with master's degrees or above (13.8% vs. 0.0%, P<0.001), a higher proportion of frontline emergency medical staff with over three years of clinical experience (100.0% vs. 91.5%, P=0.001), and more doctors (19 vs. 15, P=0.039) and nurses (46 vs. 32, P=0.039). In terms of quality and safety and medical service capacity, Zhejiang had a higher severe trauma resuscitation success rate (92.9% vs. 83.75%, P=0.003), annual emergency surgery volume (1 002 vs. 428, P=0.015), and number of emergency surgery procedures (125 vs. 42, P=0.027), but Jiangsu had a shorter emergency room stay time (87.5 min vs. 136.2 min, P=0.029). In terms of informatization, Zhejiang outperformed Jiangsu in all indicators, especially in patient information interconnection and sharing (100.0% vs. 82.4%, P=0.030) and in-hospital diagnosis and treatment timeline (91.7% vs. 73.5%, P=0.043). Correlation analysis showed that the total specialty scale score was significantly positively correlated with the in-hospital emergency rescue service capacity score ( r=0.576) and the staffing score ( r=0.455). The total evaluation score was significantly positively correlated with the informatization total score ( r=0.397) and the pre-hospital emergency rescue service capacity score ( r=0.322). Binary Logistic regression analysis showed that the emergency rescue service capacity score was significantly positively correlated with hospital grade ( B=1.431, P=0.03, OR=4.185, 95% CI:1.152-15.205). For every 1-unit increase in the emergency department area ( B=0.002, P=0.05), the probability of a hospital being upgraded to B++ or above increased by 0.2% ( OR=1.002, 95% CI:1.000-1.004). Conclusions:Both provinces exhibit distinct advantages in emergency department development. Departmental scale and configuration significantly impact emergency service capabilities, while staffing and quality management constitute core elements for improvement. Informatization construction notably enhances emergency service efficiency and quality.

AIM:To establish methods for the isolation,identification,primary culture and evaluation of gas-tric smooth muscle cells(GSMC)from rat and mouse.METHODS:Thirty SD rats and thirty C57BL/6 mice were random-ly assigned into three groups,with 10 rats or mice in each group.Three parallel experiments were conducted to isolate pri-mary GSMC under aseptic conditions.The isolated cells were identified through morphological observation,immunofluo-rescence and Western blot.Flow cytometry was employed to analyze the purity of the cells.Additionally,trypan blue ex-clusion test was utilized to evaluate the viability of the cells after resuscitation.RESULTS:Isolated rat and mouse GSMC exhibited a fusiform or polygonal morphology.Immunofluorescence and Western blot results demonstrated the expression of α-smooth muscle actin(α-SMA).Flow cytometry showed that the positive expression rate of α-SMA was 99.6%in rat GSMC and 97.4%in mouse GSMC.Moreover,the viability rates of rat and mouse GSMC after cryopreservation were found to be greater than 97%.CONCLUSION:A stable and reliable method for the isolation,culture and evaluation of primary GSMC from rat and mouse has been established.

AIM:To establish methods for the isolation,identification,primary culture and evaluation of gas-tric smooth muscle cells(GSMC)from rat and mouse.METHODS:Thirty SD rats and thirty C57BL/6 mice were random-ly assigned into three groups,with 10 rats or mice in each group.Three parallel experiments were conducted to isolate pri-mary GSMC under aseptic conditions.The isolated cells were identified through morphological observation,immunofluo-rescence and Western blot.Flow cytometry was employed to analyze the purity of the cells.Additionally,trypan blue ex-clusion test was utilized to evaluate the viability of the cells after resuscitation.RESULTS:Isolated rat and mouse GSMC exhibited a fusiform or polygonal morphology.Immunofluorescence and Western blot results demonstrated the expression of α-smooth muscle actin(α-SMA).Flow cytometry showed that the positive expression rate of α-SMA was 99.6%in rat GSMC and 97.4%in mouse GSMC.Moreover,the viability rates of rat and mouse GSMC after cryopreservation were found to be greater than 97%.CONCLUSION:A stable and reliable method for the isolation,culture and evaluation of primary GSMC from rat and mouse has been established.

Objective:To evaluate the role of aquaporin 4 (AQP4) in dexmedetomidine-induced reduction of blood-brain barrier permeability in mechanically ventilated mice and the relationship with protein kinase C (PKC).Methods:One hundred and fifty clean-grade healthy male C57BL6 mice, weighing 20-25 g, aged 8-12 weeks, were divided into 5 groups ( n=30 each) using a random number table method: control group (group C), mechanical ventilation group (group V), LY317615 group (group L), dexmedetomidine group (group D), and dexmedetomidine+ PMA group (group DP). Group C spontaneously breathed air for 6 h. The animals were mechanically ventilated for 6 h in group V. PKC inhibitor LY3176 15 μg/kg was intraperitoneally injected at 30 min before mechanical ventilation in group L. Dexmedetomidine 50 μg/kg was intraperitoneally injected at 30 min before mechanical ventilation in D and DP groups. PKC activator PMA 15 μg/kg was intraperitoneally injected at 60 min before mechanical ventilation in group DP. Mice were anesthetized at 1 day after mechanical ventilation, then sacrificed and hippocampal tissues were taken for microscopic examination of pathological changes in the hippocampal CA1 and CA3 areas (with a light microscope). Brain tissues were also taken to measure the water content and content of Evans blue (EB) and to detect the expression of PKC and AQP4 (by Western blot). The cognitive function was evaluated using a novel object recognition task at 3 days after mechanical ventilation. Results:Compared with group C, the water content and EB content of brain tissues were significantly increased after mechanical ventilation, the expression of PKC and AQP4 in brain tissues was up-regulated, the percentage of novel object exploration and discrimination index were decreased ( P<0.05), and the histopathological damage in the hippocampal CA1 and CA3 areas was aggravated in group V and group DP. Compared with group V, the water content and EB content of brain tissues were significantly decreased after mechanical ventilation, the expression of PKC and AQP4 in brain tissues was down-regulated, the percentage of novel object exploration and discrimination index were increased ( P<0.05), and the histopathological damage in the hippocampal CA1 and CA3 areas was significantly attenuated in group D and group L. Compared with group D, the water content and EB content of brain tissues were significantly increased after mechanical ventilation, the expression of PKC and AQP4 in brain tissues was up-regulated, the percentage of novel object exploration and discrimination index were decreased ( P<0.05), and the histopathological damage in the hippocampal CA1 and CA3 areas was aggravated in group DP. Conclusions:AQP4 is involved in dexmedetomidine-induced reduction of blood-brain barrier permeability in mechanically ventilated mice, and the mechanism is related to inhibiting activation of PKC.

Objective:To compare 2 modalities of Kirschner pin fixation for severely displaced (Campbell type Ⅱ) juxta-epiphyseal fracture of the proximal phalanx in children.Methods:A retrospective study was conducted to analyze the clinical data of 69 children with severely displaced (Campbell type Ⅱ) juxta-epiphyseal fracture of the proximal phalanx who had been treated at Department of Orthopaedics, Shenzhen Children's Hospital from January 2018 to December 2020. Clinical data: 41 boys and 28 girls; (7.6±3.2) years in age; 34 left hands and 35 right hands affected; 4 thumbs, 4 middle fingers, 5 ring fingers, and 56 little fingers injured. The patients were divided into 2 groups according to surgical modalities. Group A (35 fingers in 35 cases) was treated by closed reduction, longitudinal intramedullary fixation with a single Kirschner pin, and external plaster fixation while group B (34 fingers in 34 cases) by closed reduction, crossing fixation with 2 Kirschner pins, and external plaster fixation. The 2 groups were compared in terms of preoperative fracture angulation, operation time, postoperative fracture angulation, hospitalization days, and postoperative functional recovery.Results:There was no statistically significant difference in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). All fractures got united after 3 to 6 weeks. There was no statistically significant difference between the 2 groups in terms of preoperative fracture angulation, operation time, or postoperative fracture angulation ( P>0.05). The hospitalization days in group A were significantly shorter than that in group B ( P<0.05). Fifty children (24 cases in group A and 26 cases in group B) were followed up for (54.3±10.5) months. By the trial criteria for upper limb functional evaluation of the Hand Surgery Society, the hand function was evaluated at the last follow-up as excellent in 24 cases in group A, giving an excellent rate of 100% (24/24), and as excellent in 26 cases in group B, giving an excellent rate of 100% (26/26) too, showing no significant difference between the 2 groups ( P<0.05). No postoperative complications such as Kirschner wire breakage, needle tract infection, significant angular deformity, rotational deformity, or bone bridge formation occurred in any of the patients. Conclusion:For children with severely displaced (Campbell type Ⅱ) juxta-epiphyseal fracture of the proximal phalanx, longitudinal intramedullary fixation with a single Kirschner pin and crossing fixation with 2 Kirschner pins may achieve comparable efficacy in terms of fracture stability and function, but the former may result in shorter hospitalization days.

Objective:To study the correlation between zinc finger protein A20 (A20) and basic fibroblast growth factor (BFGF) and liver fibrosis in chronic hepatitis B.Methods:A retrospective study was conducted to select 120 patients with chronic hepatitis B diagnosed and treated in the Fifth Hospital of Shijiazhuang City from January 2019 to December 2020, all of whom underwent liver tissue biopsy, and 25 cases of liver pathological specimens who underwent liver hemangioma resection were selected. The correlation between the expression of A20 and BFGF in liver tissue and the stage of hepatic fibrosis and inflammation were analyzed.Results:The expression of A20 in the S1 - S4 phase was higher than that in the S0 phase: (6.12 ± 1.22)%, (10.18 ± 2.43)%, (16.94 ± 5.06)%, (25.99 ± 7.57)% vs. (0.81 ± 0.29)%; the expression of BFGF in the S1 - S4 phase was higher than that in the S0 phase: (6.12 ± 1.22)%, (10.18 ± 2.43)%, (16.94 ± 5.06)%, (25.99 ± 7.57)% vs. (0.81 ± 0.29)%, there were statistical differences ( P<0.05). The expression of A20 in the G1 - G4 phase was higher than that in the G0 phase: (6.56 ± 1.87)%, (10.01 ± 3.29)%, (15.54 ± 5.01)%, (25.86 ± 8.02)% vs. (0.85 ± 0.71)%; the expression of BFGF in the G1 - G4 phase was higher than that in the G0 phase: (5.91 ± 1.52)%, (9.65 ± 2.48)%, (15.03 ± 4.86)%, (24.62 ± 7.22)% vs. (0.79 ± 0.41)%, there were statistical differences ( P<0.05). The results of Pearson test showed that there was a positive correlation between liver A20 and BFGF ( r = 0.824, P<0.05). Conclusions:The expressions of A20 and BFGF in liver tissue increase with the aggravation of liver pathological fibrosis and inflammation, which can be used as important indicators to evaluate the severity of liver fibrosis.

Objective:To evaluate the healthcare accessibility and quality of diagnosis and treatment services of Internet hospitals in China.Methods:One hundred and eighty Internet hospitals in 60 cities were seleted based on the sampling of development levels in the eastern, central and western regions of China. From April to May 2023, standardized patients methodology was applied to evaluate the accessibility(including the number of Internet hospitals, functional settings, online doctor status, the doctor′s attending rate and consultation fees) and diagnosis and treatment service quality(including the diagnosis and treatment services quality, response speed and patient′s evaluation) of Internet hospitals.Results:The average opening rate of Internet hospitals in China was 52.9% (560/1 058), the average online rate of doctors was 64.2% (1 099/1 713), the average doctor′s attending rate was 33.6% (112/333), the average consultation fee was 4.85 yuan, the average score of consultation was 1.92 out of 9, the average score of diagnosis and treatment was 1.12 out of 4, the average score of the response speed was 1.70 out of 3, and patient satisfaction was 2.73 out of 3.Conclusions:The Internet hospital accessibility in China is unevenly developed, and the overall quality of diagnosis and treatment is low. It is recommended to accurately position and optimize the function of Internet hospital, establish the incentive mechanism for online consultation doctors, construct and improve the regulatory system of Internet hospital diagnosis and treatment, so as to improve the accessibility and quality of diagnosis and treatment of Internet hospitals.

Objective:To investigate the effect of early protein supplementation on the clinical outcomes of elderly ICU patients with critically ill.Methods:The study was a post-hoc analysis of a multicenter, cluster randomized controlled trial (NEED trial), which aimed to evaluate the impact of feeding protocol on nutritional implementation and outcomes in ICU patients. It was planned to include elderly patients aged ≥70 years from the NEED trial, and patients who had not started nutritional therapy by the Day 3 after enrolment, stayed in the ICU less than 7 days, missing the primary outcome were excluded. The primary outcome of this study was 28-day mortality of enrolment. Patients were categorized into Q1 (<0.6 g/kg/d), Q2 (0.6-0.83 g/kg/d), and Q3 (≥0.83 g/kg/d) groups according to the tertiles of protein supply. The log-rank test was used to compare the Kaplan-Meier survival curves for 28-day mortality. The associations between different protein groups and 28-day mortality were tested by Cox proportional hazards regression models. Subgroup analysis was conducted in patients with high (mNUTRIC score≥5) nutritional risk or patients with baseline acute kidney injury.Results:A total of 789 elderly (≥70 years) patients was included in the study, with a mean protein amount of 0.69 (0.53, 0.91) g/(kg·d) during days 3-7 after ICU admission, and mean protein amounts in the Q1 low-protein group, the Q2 medium-protein group, and the Q3 high-protein group were 0.46 (0.36, 0.53), 0.69 (0.63, 0.76), and 1.03 (0.91, 1.23) g/(kg·d), respectively. The results showed that the medium protein group associated with lower 28-day mortality compared to the high protein group, and the association between the medium protein group and lower 28-day mortality still held after controlling for possible confounders by Cox multivariate regression analysis. In the high-nutritional risk subgroup (mNUTRIC≥5), a significant association was also found between the medium protein group and lower 28-day mortality.Conclusions:Early high protein supply are not beneficial for elderly ICU patients by this large sample size post-hoc analysis, and medium protein supply associate with lower 28-day mortality compared with the high protein group. This study may provide a theoretical basis for the optimal dose of early protein supply in elderly ICU patients, as well as a reference for clinical implementation.

Objective:To evaluate the role of transient receptor potential vanillic acid 4 (TRPV4) in dexmedetomidine-induced improvement in cognitive function in mice with mechanical ventilator-caused brain injury.Methods:Ninety clean-grade healthy male C57BL6 mice, weighing 20-25 g, aged 8-12 weeks, were divided into 5 groups ( n=18 each) using a random number table method: control group (group C), mechanical ventilation group (group V), HC-067047 group (group H), dexmedetomidine group (group D), and dexmedetomidine+ GSK1016790A group (group DG). In group C, the animals breathed air spontaneously for 6 h without mechanical ventilation. In group V, the animals were mechanically ventilated for 6 h. In group H, TRPV4 blocker HC-067047 10 mmol was injected into the cerebral ventricle at 3 and 6 h of mechanical ventilation. In D and DG groups, dexmedetomidine 50 μg/kg was intraperitoneally injected at 30 min before mechanical ventilation. In group DG, TRPV4 agonist GSK1016790A 5 μmol was injected into the cerebral ventricle at 60 min before mechanical ventilation. Morris water maze test was performed on 6 mice in each group at 1 day before mechanical ventilation and 3 and 7 days after mechanical ventilation. Six mice in each group were randomly selected and sacrificed at 1 day after mechanical ventilation, and the brain tissue was taken for determination of the neuronal apoptosis in hippocampal CA1 area by TUNEL method, and the apoptosis index was calculated. Six mice in each group were randomly selected and sacrificed at 1 day after mechanical ventilation, and the hippocampal tissues were taken for determination of the expression of TRPV4, serine-threonine protein kinase (Akt), phosphorylated Akt (p-Akt), Bcl-2, Bax and caspase-3 by Western blot. Results:Compared with group C, the escape latency was significantly prolonged and the number of crossing the original platform was reduced at 3 and 7 days after mechanical ventilation, the expression of TRPV4 and caspase-3 was up-regulated, the ratio of Bcl-2/Bax was decreased, and the apoptosis index of neurons was increased in group V and group DG ( P<0.05). Compared with group V, the escape latency was significantly shortened and the number of crossing the original platform was increased at 3 and 7 days after mechanical ventilation, the expression of TRPV4 and caspase-3 was down-regulated, the expression of p-Akt was up-regulated, the ratio of Bcl-2/Bax was increased, and the apoptosis index of neurons was decreased in group D and group H ( P<0.05). Compared with group D, the escape latency was significantly prolonged at 3 and 7 days after mechanical ventilation, the number of crossing the original platform was reduced, the expression of TRPV4 and caspase-3 was up-regulated, the expression of p-Akt was down-regulated, the ratio of Bcl-2/Bax was decreased, and the apoptosis index of neurons was increased in group DG ( P<0.05). Conclusions:TRPV 4 is involved in dexmedetomidine-induced improvement in cognitive function, which is related to up-regulation of p-Akt expression and inhibition of apoptosis in hippocampal neurons in mice with mechanical ventilation-caused brain injury.