BACKGROUND:Bone relies on the close connection between blood vessels and bone cells to maintain its integrity.Bones are in a physiologically hypoxic environment.Therefore,the study of angiogenesis and osteogenesis in hypoxic environment is closer to the microenvironment in vivo. OBJECTIVE:To explore the influence of Wenshen Tongluo Zhitong(WSTLZT)formula on H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell co-culture system in hypoxia environment and its related mechanism. METHODS:Enzyme digestion method and flow sorting technique were used to isolate and identify H-type bone microvascular endothelial cells.Mouse bone marrow mesenchymal stem cells were isolated and obtained by bone marrow adhesion method.H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell hypoxic co-culture system was established using Transwell chamber and anoxic culture workstation.WSTLZT formula powder was used to intervene in each group at a mass concentration of 50 and 100 μg/mL.The angiogenic function of H-type bone microvascular endothelial cells in the co-culture system was evaluated by scratch migration test and tube formation test.The osteogenic differentiation ability of bone marrow mesenchymal stem cells in the co-cultured system was evaluated by alkaline phosphatase staining and alizarin red staining.The protein and mRNA expression changes of PDGF/PI3K/AKT signal axis related molecules in H-type bone microvascular endothelial cells in the co-cultured system were detected by Western Blotting and q-PCR,respectively. RESULTS AND CONCLUSION:(1)Compared with the normal oxygen group,the scratch mobility and new blood vessel length of H-type bone microvascular endothelial cells were significantly higher(P<0.05);the osteogenic differentiation capacity of bone marrow mesenchymal stem cells was higher(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular protein and mRNA increased(P<0.05)in the hypoxia group.(2)Compared with the hypoxia group,scratch mobility and new blood vessel length were significantly increased in the H-type bone microvascular endothelial cells(P<0.05);bone marrow mesenchymal stem cells had stronger osteogenic function(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular proteins and mRNA further increased(P<0.05)after treatment with different dose concentrations of WSTLZT formula.These findings conclude that H-type angiogenesis and osteogenesis under hypoxia may be related to the PDGF/PI3K/AKT signaling axis,and WSTLZT formula may promote H-type vasculo-dependent bone formation by activating the PDGF/PI3K/AKT signaling axis,thereby preventing and treating osteoporosis.

【Objective】 To compare the effects of 3 rehydration methods before blood donation on the prevention of on-site and delayed blood donation-related vasovagal response (VVR) . 【Methods】 From January to June 2021, 6 250 whole blood donors in 6 fixed blood donation sites signed informed consent and were divided into 198 clusters according to donor sites and dates, then they were randomly assigned to receive either oral rehydration salts (ORS), sugar water, or water group, and each drank 500 mL of ORS, sugar water or water within 20 minutes before blood donation. The researchers recorded the actual intervention accepted on site, and recorded the immediate VVR and related information. At rest after blood donation, donors submitted an electronic questionnaire containing socio-demographic information. At 48 hours after blood donation, the researchers called back every donor to record delayed VVR and related information. Logistic regression based on intention to treat (ITT) was used to analyze the difference of the incidence of VVR among the three groups, and the average treatment effect on treated (ATT) was calculated. PASS 2021was used to estimate the sample size and R (4.2.0) for statistical analysis. 【Results】 The cumulative incidence of blood donation-related VVR was 2.67% (2.29%-3.11%) among street whole blood donors under the 3 rehydration methods, in which, the incidence of immediate and delayed VVR was 1.02% (0.79%-1.31%) and 1.65% (1.36%-2.01%) respectively. ITT analysis found that ORS were more effective than water in reducing the incidence of delayed VVR【OR=0.59,95% CI[0.37,0.94]】.There was no significant difference in the incidence of immediate VVR between any two groups (P > 0.05), and there was no significant difference in the incidence of delayed VVR in the sugar water group compared with the water group (P > 0.05). There was a difference of -0.013 (【95% CI[-0.022, -0.004]】or -0.008【95% CI[-0.017, -0.000]】in the incidence of delayed VVR in the ORS group compared with water group or sugar water group, the difference was significant (P<0.05). The cumulative VVR of the three groups showed similar results to the delayed VVR. 【Conclusion】 Drinking ORS before blood donation is the most effective rehydration method to prevent delayed VVR. The next step is to establish the predictive model of delayed VVR to screen the susceptible population and provide them with ORS before blood donation, while other population can choose any liquid they like, thus achieving personalized blood donation-related VVR prevention and control.

【Objective】 To explore the model of "One Core, Multiple Elements" emergency blood donation volunteer team (referred to as the Model) . 【Methods】 The Nanjing City Voluntary Blood Donation Joint Meeting serves as the core, with diverse entities including party committees, government departments, district governments, social organizations, enterprises, blood donors, etc. Following the principles of "emergency response in emergencies, wartime readiness, combining regular and wartime efforts," and adhering to the framework of the Model, the emergency blood donation volunteer team system in Nanjing was constructed. 【Results】 1) After the construction of the Model (2018—2022), the total number of emergency blood donation volunteers in Nanjing City increased by 191% compared to the pre-construction period (2013—2017), with an average annual blood donation of 20 929, showing significant differences (P<0.05). 2) After the construction of the Model, the number of emergency blood donation during the winter and summer increased by 206% and 185%, respectively, compared to the pre-construction period, demonstrating significant differences (P< 0.05) .3) Prior to the construction of the Model, Nanjing lacked a relatively stable emergency volunteer team. After the construction, Nanjing established a total of 5 relatively stable volunteer teams, with the district-level government and township personnel accounting for the highest proportion (52.96%), followed by medical personnel (23.95%), enterprise team (11.10%), State-owned Assets Supervision and Administration Commission team (7%), and Nanjing municipal government team (4.98%). 4) Following the Model, during the COVID-19 pandemic, Nanjing successfully initiated two Level Ⅱ blood emergency responses, with emergency blood donations reaching 23 041. 【Conclusion】 The Model can effectively ensure the blood supply in Nanjing region during emergencies.

Objective To analyze the epidemic status of newly diagnosed occupational diseases in Guangdong Province during the “13th Five-Year Plan” period, and scientifically evaluate the critical risk factors for occupational disease prevention and control. Methods The data of newly diagnosed occupational diseases reported by internet in Guangdong Province from 2016 to 2020 was collected from Report Card of Occupational Diseases using Occupational Diseases and Health Hazard Factors Monitoring Information System under China Disease Prevention and Control Information System. Data was used to determine the epidemic status and identify key risk factors. Results A total of 4 846 cases of occupational diseases were reported in Guangdong Province during the “13th Five-Year Plan”period, with an increase rate of 39.13% compared with the “12th Five-Year Plan” period (3 483 cases). The annual distribution of newly diagnosed occupational disease cases showed a trend of increasing and then declining. The top five percentage on types of occupational diseases were occupational noise-induced deafness, occupational silicosis, occupational chronic benzene poisoning, other occupational pneumoconiosis and occupational hand-arm vibration disease, accounting for 45.23%, 16.28%, 11.52%, 7.92% and 4.60%, respectively. Cases from the Pearl River Delta region accounted for 92.76%, while five non-Delta cities had “zero reported cases”. Regarding industry sector, cases were primarily concentrated in manufacturing, construction, and mining, accounting for 84.21%, 5.49% and 3.59%, respectively. Regarding industry categories, cases were concentrated in metal products, non-metallic mineral products, manufacturing of cultur, education, art, sports and entertainment goods, manufacturing of computer, communication and other electric devices, and manufacturing of specialized equipment, accounting for 11.70%, 10.17%, 8.60%, 7.82%, and 4.81%, respectively. A total of 196 enterprises (accounting for 7.39%) reported an average of three or more new cases per year, while 19 enterprises (accounting for 0.72%) reported an average of ten or more cases per year. Conclusion The epidemic status of occupational diseases in Guangdong Province showed a “triple concentration” characteristic in disease types, regions, and industries during the “13th Five-Year Plan” period. Particular attention should be paid to epidemic status of occupational noise-induced deafness, occupational silicosis, occupational chronic benzene poisoning, other occupational pneumoconiosis, and occupational hand-arm vibration disease, and also pay special attention to the prevention and control of occupational diseases in the Pearl River Delta region, cities with “zero reported cases”, manufacturing industry and occupational disease-prone enterprises.

Objective To investigate the effects of zalcitabine(ddC),a mitochondrial DNA polymerase γ(POLG)inhibitor,on the migration,invasion,and to preliminarily explore mitochondrial biogenesis of human tri-ple-negative breast cancer MDA-MB-231 cells.Methods The effect of ddC on cell viability was detected using the MTT assay.The migration and invasion abilities of the cells were evaluated using the cell scratch and Transwell in-vasion assays.Cell apoptosis was determined using flow cytometry and a V-FITC/PI cell apoptosis detection kit.The protein expression of POLG,NADH dehydrogenase subunit Ⅰ(NADH1),NADH dehydrogenase subunit Ⅱ(NADH2),ATP synthase subunit 6(ATPase6),cytochrome c oxidase subunit Ⅰ(COX-1)and cytochrome c ox-idase subunit Ⅲ(COX-3)were determined using Western blot.The POLG mRNA level and mtDNA copy number were determined using qPCR.The mitochondrial content and ATP levels were determined using MitoTracker Green fluorescent probe staining and an ATP determination kit.MDA-MB-231 cells were transfected with pcDNA3.1-EG-FP-POLG plasmids to overexpress POLG.The inhibitory effects of ddC on cell migration and invasion were detected in POLG-overexpressed MDA-MB-231 cells.Results POLG expression was higher in MDA-MB-231 cells than in normal mammary epithelial cells(MCF-10A)(P＜0.01).ddC inhibited cell viability in a dose-dependent man-ner.ddC inhibited the migration(P＜0.01)and invasion(P＜0.01)of MDA-MB-231 cells;however,it dis-played no significant inhibitory effects on cell viability in normal mammary epithelial cells(MCF-10A)at the same concentration.ddC downregulated the protein(P＜0.01)and mRNA(P＜0.01)levels of POLG,reduced mtD-NA copy number(P＜0.01)and downregulated mtDNA-coded NADH1,NADH2,ATPase6,COX-1 and COX-3 protein expression(P＜0.01)in MDA-MB-231 cells.Furthermore ddC inhibited mitochondrial content(P＜0.01)and ATP(P＜0.01)levels in MDA-MB-231 cells.POLG overexpression increased the migration(P＜0.05)and invasion(P＜0.05)abilities of MDA-MB-231 cells,while ddC did not significantly inhibit the migra-tion and invasion abilities of MDA-MB-231 cells overexpressing POLG.Conclusion ddC downregulates POLG ex-pression in MDA-MB-231 cells and inhibits mitochondrial biogenesis and ATP levels,thereby inhibiting the migra-tion and invasion of MDA-MB-231 cells.

Objective:To construct a mouse asthma model induced by mugwort pollen and to explore endotyping,providing methods for subsequent precision treatment.Methods:BALB/c mice were intraperitoneally injected with mugwort pollen extract(MPE)to sensitize,following MPE intranasal challenge to construct MPE allergic asthma murine model.Mice were randomly divided into PBS sensitization and PBS challenge(P-P),MPE sensitization and PBS challenge(M-P),MPE sensitization and MPE challenge model(M-M)groups.24 h after final challenge,mice were performed to examine airway responsiveness;bronchoalveolar lavage fluid(BALF)was harvested for cell counting and statistical classification of inflammatory cells through flow cytometry analysis.Pulmonary slides were collected for pathological examination,including HE,PAS,Masson and α-SMA immunohistochemical staining.ELISA was used to detect levels of IFN-γ,IL-4,IL-5,IL-13,IL-17A in lung tissue and serum,as well as serum total IgE and MPE-specific IgE,IgG1,IgG2a levels.Results:Pathological examination showed higher airway reactivity,more inflammatory cells infiltration around airway,obvious goblet metaplasia,thickening of airway smooth muscles and dramatical fibrin deposition around airway in model group.Total cell numbers of BALF were increased from<1×105 cells/ml in P-P group to>5×105 cells/ml in model group,in which eosinophils were predominant cellular type,levels of IL-4,IL-13,IL-17A in lung and IL-5,IL-13 levels in serum were significantly increased,as well as significant increasing levels of total IgE and MPE-specific IgE,IgG1,IgG2a.Conclusion:MPE-sensitized and challenged mice induces typical eosinophilic asthma featured with elevated eosinophils,as well as secretion of inflammatory factors of type 2 and type 17,IgE,IgG1 and IgG2a subtypes soars to high levels.

Objective:To investigate the clinical value of deep learning model based on contrast enhanced ultrasound (CEUS) video in the differential diagnosis of benign and malignant liver tumors.Methods:Between May 2010 and June 2022, 1 213 patients who underwent CEUS examination for liver masses in the Affiliated Hospital of Southwest Medical University were retrospectively collected, and the enrolled patients were divided into training and independent test cohorts with December 31, 2021 as the time cut-off. In the training cohort, the TimeSformer algorithm was used as the infrastructure, and multiple fixed-time segments were obtained from CEUS arterial videos by using the sliding window of the video, and the classification results of the entire video were obtained after fusing the features of multiple segments, so as to build a deep learning model based on CEUS videos. In the independent test cohort, ROC curves were used to verify the validity of the model and compared with three radiologists with different CEUS experience (R1, R2, and R3, with 3, 6, and 10 years of CEUS experience, respectively).Results:A total of 1 213 patients with liver masses were included in the study, including 1 066 patients in the training cohort (426 cases of malignancy) and 147 patients in the independent test cohort (50 cases of malignancy). The area under curve (AUC)value of deep learning model was 0.93±0.01 in the training cohort and 0.89±0.01 in the independent test cohort, and the accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were 80.42%, 74.19%, 92.00%, 94.52% and 65.71%, respectively. Among the three radiologists, R1 had the lowest diagnostic performance, with accuracy, sensitivity, specificity, PPV and NPV of 67.83%, 51.61%, 98.00%, 97.96% and 52.13%, respectively, while the above indicators of R3 were 82.52%, 76.36%, 94.00%, 95.95% and 68.12%, respectively. McNemar′s test showed that the difference between R1 and the deep learning model was statistically significant ( P<0.001), while the differences between R2 and R3 and the deep learning model were not statistically significant ( P=0.720, 0.868). In addition, the analysis time of the model for a single case was (340.24±16.32)ms, while the average analysis time of radiologists was 62.9 s. Conclusions:The deep learning model based on CEUS can better identify benign and malignant liver masses, and may reach the diagnostic level of experienced radiologists.

OBJECTIVE To study the pharmacokinetics of Esketamine hydrochloride nasal spray in rats and ciliary toxicity to maxillary mucosa of bullfrog. METHODS The plasma concentration of esketamine hydrochloride in rats was determined by LC-MS/ MS after intravenous injection of esketamine hydrochloride solution and nasal administration of esketamine hydrochloride； the pharmacokinetic parameters were calculated by using Phoenix WinNonlin 8.1.0 software. Using the maxillary mucosa of isolated bullfrog as a model， the morphological changes of maxillary mucosa were investigated， and the duration and recovery of ciliary oscillation were recorded after nasal administration of esketamine hydrochloride. RESULTS The peak of blood concentration occurred 2 min after nasal administration of esketamine hydrochloride； cmax was （814.58±418.80） ng/mL， AUC0－∞ was （203.75± 92.76） ng·h/mL， and the absolute bioavailability was 60.68%. After nasal administration of esketamine hydrochloride， it was observed that the cilia of bullfrog were arranged neatly， the edges were clear， the cilia tissue structure was complete and the cilia moved actively. The cilia movement time was （178.17±13.30） min for the first time， and after the cilia moved again， the ciliary movement time measured again was （24.50±9.19）min with a relative movement percentage of 53.56%. CONCLUSIONS Esketamine hydrochloride nasal spray has a rapid onset of action， high bioavailability， and low ciliary toxicity.

Objective To investigate the prevalence of Echinococcus infections in small rodents around human residential areas in Yushu City, Qinghai Province in 2023, so as to provide insights into precision echinococcosis control. Methods One or two quadrats, each measuring 50 m × 50 m, were randomly assigned in Shanglaxiu Township and Longbao Township, Yushu City, Qinghai Province on June 2023, respectively, and 300 plate-type mouse traps, each measuring 12.0 cm × 6.5 cm, were assigned in each quadrat. Small rodents were captured during the period between 10 : 00 and 18 : 00 each day for 4 days. Then, all captured small rodents were identified and dissected, and liver specimens with suspected Echinococcus infections were subjected to pathological examinations. The Echinococcus cytochrome c oxidase 1 (cox1) gene was amplified using PCR assay, and the sequence of the amplified product was aligned to that was recorded in the GenBank to characterize the parasite species. In addition, a phylogenetic tree of Echinococcus was generated based on the cox1 gene sequence using the neighbor-joining method. Results A total of 236 small rodents were captured in Shanglaxiu and Longbao townships, Yushu City, including 65 Qinghai voles and 51 plateau pikas in Shanglaxiu Township, and 62 Qinghai voles and 58 plateau pikas in Longbao Township, and there was no significant difference in the constituent ratio of small rodents between the two townships (χ² = 0.294, P > 0.05). Seven plateau pikas and 12 Qinghai voles were suspected to be infected with Echinococcus by dissection, and pathological examinations showed unclear structure of hepatic lobules and disordered hepatocyte arrangement in livers of small rodents suspected of Echinococcus infections. PCR assay identified E. shiquicus DNA in 7 Qinghai voles, which were all captured from Shanglaxiu Township. Phylogenetic analysis showed that the cox1 gene sequence of Echinococcus in small rodents was highly homologous to the E. shiquicus cox1 gene sequence reported previously. Conclusion Plateau pika and Qinghai vole were predominant small rodents around human residential areas in Yushu City, Qinghai Province in 2023, and E. shiquicus infection was detected in Qinghai voles.

Objective: To investigate the molecular typing and drug resistance of Salmonella in market-sold raw livestock meats in Shaoxing City, Zhejiang Province, so as to provide the basis for the prevention and control of drug-resistant Salmonella. Methods: Salmonella isolates were detected and serotyped from raw livestock meat samples collected from supermarkets, agricultural markets and restaurants in Shaoxing City between March and November 2023. Molecular typing was performed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Antibiotic resistance phenotypes and resistance gene analyses were conducted using broth microdilution method and whole genome sequencing. Results: A total of 34 Salmonella isolates were detected from 329 samples and the positive rate was 10.33%. There were 14 serotypes, with S. Rissen (9 isolates, 26.47%) and S. London (8 isolates, 23.53%) being dominant. PFGE typing revealed 31 band patterns and 23 banding pattern clusters of Salmonella, and the similarity ranged from 30.80% to 100.00%, without dominant band patterns. MLST identified 15 sequence typing (ST) types, and S. Rissen were all ST469 and S. London were all ST155. Salmonella strains exhibited high levels of resistance to tetracycline (44.12%) and ampicillin (35.29%). Thirteen resistance patterns were identified, with 8 multidrug resistant patterns (29.41%). There were 8 classes of 14 resistance genes, with aminoglycoside resistance gene aac(6' )-Iaa found in 100.00% of the strains. The highest concordance rate between the resistance phenotype and gene was observed for phenicol antibiotics (97.06%), while there was no concordance between the quinolone resistance phenotype and genes. Conclusions S. Rissen and S. London are the predominant serotypes in market-sold raw livestock meats in Shaoxing City. The PFGE band types of the Salmonella isolates show polymorphism distribution, and MLST are mainly ST469 and ST155. The isolates have high levels of resistance to tetracycline and ampicillin, manifesting multidrug resistance.