Natural products (NPs) are an important source of new drugs for the treatment of stroke. Identifying cellular targets for bioactive molecules is a major challenge and critical issue in the development of new drugs for stroke. Small-molecule probes play a unique role in target discovery. However, drawbacks to these probes include non-specificity, unstable activity, and difficulty in synthesis. Small-molecule probes based on NPs at least partially compensate for these shortcomings. NPs feature rich chemical and structural diversity, biocompatibility, and unique biological activities. These features could be exploited to provide new ideas and tools for target discovery. Small-molecule probes based on NPs provide a precise and direct search for interacting protein targets of NPs-active small molecules. This review explores the properties of small-molecule probes based on NPs and their applications in mechanistic studies of stroke and other diseases. We hope that this review will bring new perspectives to the mechanistic study of NPs-active small molecules and accelerate the translation of these ingredients into drug candidates for the treatment of stroke.

Purpose To investigate the expression of Versican(VCAN)and thrombospondin 2(THBS2)and their relationship with cancer-associated fibroblast(CAFs)and clinicopathological significance in papillary thyroid car-cinoma(PTC).Methods Bioinformatics analyses were performed using PTC single-cell sequencing data from the GEO and TCGA database.Weighted correlation network analysis identified CAFs-related genes,and enrichment analy-sis highlighted pivotal genes associated with CAFs;the expression of VCAN,THBS2,and α-SMA in 130 PTC tissue samples were detected by immunohistochemistry EnVision method.Masson staining evaluated tumor stromal fibrosis.Relationships between these markers,clinicopathological parameters,and CAF proliferation were analyzed.Results Bioinformatics analysis identified VCAN and THBS2 as core genes significantly associated with CAFs,and extracellular matrix-related pathways.The proliferation rate of CAFs in PTC was 83.1％(108/130),with positivity rate of 96.9％(126/130)for VCAN and 75.4％(98/130)for THBS2.The median mesenchymal fibrosis index was 32.4(inter-quartile range:22.7-50.0).High CAF proliferation correlated positively with lymph node metastasis(P＜0.001),higher TNM stage(P＜0.05),and specific histologic subtypes of PTC.Similarly,VCAN expression,THBS2 expres-sion,and the degree of PTC stromal fibrosis were positively correlated with lymph node metastasis(P＜0.001,P＜0.05,and P＜0.001,respectively).Both THBS2 expression and the degree of PTC stromal fibrosis correlated with the histologic subtype of PTC.The percentage of tumor mesenchymal α-SMA-positive cells strongly correlated with the im-mune response score(IRS)of VCAN and THBS2(rs=0.713,P＜0.001;rs=0.646,P＜0.001).Additionally,the percentage of Masson-stained area was positively correlated with the percentage of tumor mesenchymal α-SMA-posi-tive cells,and the IRS of VCAN and THBS2(rs=0.892,P＜0.001;rs=0.729,P＜0.001;rs=0.616,P＜0.001).Conclusion VCAN and THBS2 serve as potential markers for assessing invasiveness and lymph node metas-tasis of PTC.Their strong association with CAFs provides a basis for further investigation of the malignant biological be-havior of PTC.

Purpose To investigate the expression of Versican(VCAN)and thrombospondin 2(THBS2)and their relationship with cancer-associated fibroblast(CAFs)and clinicopathological significance in papillary thyroid car-cinoma(PTC).Methods Bioinformatics analyses were performed using PTC single-cell sequencing data from the GEO and TCGA database.Weighted correlation network analysis identified CAFs-related genes,and enrichment analy-sis highlighted pivotal genes associated with CAFs;the expression of VCAN,THBS2,and α-SMA in 130 PTC tissue samples were detected by immunohistochemistry EnVision method.Masson staining evaluated tumor stromal fibrosis.Relationships between these markers,clinicopathological parameters,and CAF proliferation were analyzed.Results Bioinformatics analysis identified VCAN and THBS2 as core genes significantly associated with CAFs,and extracellular matrix-related pathways.The proliferation rate of CAFs in PTC was 83.1％(108/130),with positivity rate of 96.9％(126/130)for VCAN and 75.4％(98/130)for THBS2.The median mesenchymal fibrosis index was 32.4(inter-quartile range:22.7-50.0).High CAF proliferation correlated positively with lymph node metastasis(P＜0.001),higher TNM stage(P＜0.05),and specific histologic subtypes of PTC.Similarly,VCAN expression,THBS2 expres-sion,and the degree of PTC stromal fibrosis were positively correlated with lymph node metastasis(P＜0.001,P＜0.05,and P＜0.001,respectively).Both THBS2 expression and the degree of PTC stromal fibrosis correlated with the histologic subtype of PTC.The percentage of tumor mesenchymal α-SMA-positive cells strongly correlated with the im-mune response score(IRS)of VCAN and THBS2(rs=0.713,P＜0.001;rs=0.646,P＜0.001).Additionally,the percentage of Masson-stained area was positively correlated with the percentage of tumor mesenchymal α-SMA-posi-tive cells,and the IRS of VCAN and THBS2(rs=0.892,P＜0.001;rs=0.729,P＜0.001;rs=0.616,P＜0.001).Conclusion VCAN and THBS2 serve as potential markers for assessing invasiveness and lymph node metas-tasis of PTC.Their strong association with CAFs provides a basis for further investigation of the malignant biological be-havior of PTC.

Objective:To assess the value of optical genome mapping (OGM) for the detection of chromosomal structural abnormalities including ring chromosomes, balanced translocations, and insertional translocations.Methods:Clinical data of four patients who underwent pre-implantation genetic testing concurrently with OGM and chromosomal microarray analysis at the Center of Reproductive Medicine of the Sixth Affiliated Hospital of Sun Yat-sen University from January to October 2022 due to chromosomal structural abnormalities were selected as the study subjects. Some of the results were verified by multi-color fluorescence in situ hybridization. Results:The OGM has successfully detected a balanced translocation and fine mapped the breakpoints in a patient. Among two patients with insertional translocations, OGM has provided more refined breakpoint locations than karyotyping analysis in a patient who had chromosome 3 inserted into chromosome 6 and determined the direction of the inserted fragment. However, OGM has failed to detect the chromosomal abnormalit in a patient with chromosome 8 inserted into the Y chromosome. It has also failed to detect circular signals in a patient with ring chromosome mosaicism.Conclusion:OGM has successfully detected chromosomal structural variations in the four patients and provided assistance for their diagnosis.

Objective:To investigate the molecular epidemiological characteristics of coxsackievirus A16 (CVA16) in Fujian province from 2020 to 2023.Methods:The epidemiological characteristics of CVA16 associated hand, food and mouth disease (HFMD) in Fujian province from 2020 to 2023 was analyzed. The complete VP1 gene of CVA16 was amplified by RT-PCR and then sequenced, and genetic evolution was analyzed by MEGA X and other softwares.Results:From 2020 to 2023, there were 13 120 cases of HFMD in Fujian province, and the proportion of HFMD which caused by CVA16 was 16.5% (2 160/13 120). From 2020 to 2023, the proportion of accounted cases was 4.7% (94/2 019), 14.1% (457/3 243), 47.6% (1 521/3 199) and 1.9% (88/4 659) respectively. HFMD caused by CVA16 was mainly concentrated in children aged 1 to 5 years, and most of them were 3 years old. The genetic evolution and genotype analysis of 92 complete VP1 gene sequences obtained from 2020 to 2023 showed that the genetic distance between CVA16 strains in Fujian province and the prototype strain was far away. The CVA16 genotype in Fujian province from 2020 to 2023 has three clusters of B1a, B1b and B1c, among which the composition ratio of B1a and B1b in Fujian province in 2020 was 40% and 60% respectively. In 2021, B1a and B1b accounted for 81.8% and 18.2% respectively. Only B1a in 2022; in 2023, there were B1a, B1b and B1c, which respectively accounted for 44.4%, 7.4% and 48.2%. During the period from January to September, B1a was the main cluster. After October we observed an emergence of B1c cluster, which had never been found in Fujian province and was rare in China, was detected and became the dominant cluster.Conclusions:The evolutionary cluster of CVA16 dominant changed from B1b in 2020 to B1a in 2021-2023. After October 2023, the newly discovered B1c became the dominant cluster in Fujian province.

Objective To explore the effect of intermittent fasting on blood glucose control in patients with type 2 diabetes mellitus(T2DM)complicated by overweight and obesity.Methods 46 T2DM patients with overweight and obesity were divided into control group(Con,n=22)and intermittent fasting group(IF,n=24)according to their wishes.Con group was given routine diet and drug treatment by specialists in endocrinology.IF group completed the intermittent fasting by using a combination of meal replacement and natural diet,and hypoglycemic drugs were reduced on fasting days.Results Compared with Con group,IF group showed a significant decrease in body weight,BMI,hypoglycemic drug efficacy score,WC,fat index,hip circumference and visceral fat(VF)(P＜0.05).After intervention,the proportion of diabetes in remission or near remission in IF group was higher than that in Con group(P＜0.05).Conclusion Intermittent fasting can reduce the application of hypoglycemic drugs in T2DM patients with overweight and obesity,and can also reduce weight and VF.

Objective:To investigate the effect of fluid resuscitation and circulatory support, directed by different target mean arterial pressure (MAP), on abdominal blood flow, gastrointestinal function and inflammatory response in septic shock patients with hypertension.Methods:A prospective randomized controlled study was conducted. Hypertensive patients with septic shock admitted to the department of intensive care unit (ICU) of Liuzhou People's Hospital from January 1, 2019 to May 31, 2020 were enrolled. Patients were randomly divided into the low MAP groups (low standard group, LS group) or high MAP group (high standard group, HS group). According to the Surviving Sepsis Campaign Guidelines in 2016 and the updated guideline in 2018, all patients were given treatment of primary disease, fluid resuscitation, supportive management. The target MAP was 65-70 mmHg (1 mmHg = 0.133 kPa) in LS group, and was 75-80 mmHg in HS group. Acute gastrointestinal function injury (AGI) classification was performed on the 1st, 3rd and 7th day. The mean flow rate (Vm) and resistance index (RI) of superior mesenteric artery were evaluated using ultrasound, and the gastrointestinal function was dynamically evaluated using the modified single section ultrasonic gastric antrum method. The gastric antrum movement index (MI) and gastric empaging time (GET) were recorded. The levels of inflammatory markers in serum were detected by enzyme linked immunosorbent assay (ELISA), such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), procalcitonin (PCT) and vascular endothelial growth factor (VEGF). The target MAP, the days of use of vasopressors and the amount of fluid resuscitation were recorded.Results:A total of 208 hypertensive patients with septic shock were enrolled, including 109 in the LS group and 99 in the HS group. There were no significant differences in gender, age, acute physiology and chronic health evaluationⅡ (APACHEⅡ) score and sequential organ failure assessment (SOFA) score between the two groups when diagnosed. After treatment, there was no significant difference in AGI classification between the LS group and HS group on the 1st day. On the 3rd and 7th day, there were statistical differences between the two groups (3rd day: proportion of Ⅰ, Ⅱ, Ⅲ, Ⅳ grades were 25.69%, 56.88%, 11.93%, 5.50% in LS group, 15.15%, 54.55%, 25.25%, 5.05% in HS group, respectively, χ 2 = 7.900, P = 0.048; 7rd day: proportion of Ⅰ, Ⅱ, Ⅲ, Ⅳ grades were 44.96%, 49.54%, 3.67%, 1.83% in LS group, 31.31%, 52.53%, 11.11%, 5.05% in HS group, respectively, χ 2 = 8.178, P = 0.042). The Vm of superior mesenteric artery was higher and the RI was lower in the LS group than those in the HS group on day 1, 3 and 7 [Vm (cm/s): 21.72±3.02 vs. 19.50±2.83, 20.42±2.62 vs. 17.02±1.99, 26.52±2.70 vs. 22.47±4.03; RI: 0.86±0.05 vs. 0.92±0.04, 0.87±0.05 vs. 0.95±0.05, 0.81±0.03 vs. 0.85±0.03, all P < 0.01]. The MI was higher and the GET was shorter in the LS group than those in the HS group on day 3 and day 7 [MI: 3.00±0.33 vs. 2.60±0.29, 4.50±0.51 vs. 3.90±0.33; GET (minutes): 86.01±19.78 vs. 100.99±25.01, 71.00±16.37 vs. 84.98±20.18, all P < 0.01]. In addition, the levels of serum TNF-α, IL-6, PCT, VEGF were lower in the LS group than those in the HS group after 3 days of treatment [TNF-α (ng/L): 147.05±28.32 vs. 256.99±27.04, IL-6 (ng/L): 762.99±57.83 vs. 1 112.30±118.32, PCT (μg/L): 37.00±5.58 vs. 56.00±12.36, VEGF (ng/L): 123.00±19.78 vs. 167.01±21.55, all P < 0.05]. The target MAP was maintained at (68.02±4.71) mmHg in LS group, and (79.04±3.04) mmHg in HS group. The difference between the two groups was statistically significant ( P < 0.01). Compared with the HS group, the days of using vasopressors was shorter in LS group (days: 3.50±1.27 vs. 4.55±1.47), and the amountof fluid was reduced significantly (mL: 1 602.29±275.49 vs. 2 000.30±272.59, both P < 0.01). Conclusion:Maintaining a low target mean arterial pressure (65-70 mmHg) in hypertensive patients with septic shock can improve blood supply of superior mesenteric artery, protect the gastrointestinal function, reduce the level of inflammatory factors, and diminish the duration of using vasopressors and the amount of fluid.

Objective To understand the characteristics of minipigs infected withJapanese encephalitis virus(JEV).Methods After the brain tissues were treated, the pig brain tissue treatment solution was inoculated with BHK21 cells.Then, virus culture,indirect immunofluorescence assay, neutralization test, electron microscopic observation, and reverse transcription-polymerase chain reaction (RT-PCR) amplification of the new isolate E segment and PrM segment nucleotide sequence were performed and the genotype was identified.Results BHK21 cells were inoculated into 25 pigbrain tissues.Among them, three tissue-treated fluid couldinduce shrinkage and aggregation of BHK21 cells, and immunofluorescence staining showed strong green fluorescence response.The results of neutralization test showed that the neutralization titer of these three new isolates was 1:64, and the size of the virus particles was about 40nm under the electron microscope.The homology of both RT-PCR product sequencing results and E-segment of vaccine strain were 95%.Three new isolates were type GIII JEV.Conclusion The results ofthisstudydemonstrate that there is G III type Japanese encephalitis virus infection in the minipig farm.

Objective: To investigate the efficacy of 200IU hepatitis B immunoglobulin (HBIG) injection at 1 month after birth to interrupt the mother-to-children transmission (MTCT) of hepatitis B virus (HBV). Methods: Infants born to mothers who were hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) positive, with HBV DNA load ≥1.0×10⁶ IU/ml and who did not receive antiviral drug treatment during pregnancy, were randomly divided into 2 groups. Infants in the control group were treated with standard immunoprophylaxis: 200 IU HBIG and 10 μg recombinant hepatitis B vaccine injection within 2 h after birth and a vaccine booster at 1 and 6 months after birth. For infants in the HBIG group the standard immunoprophylaxis and an additional 200 IU HBIG were administered at 1 month. HBsAg, the antibody to HBsAg (anti-HBs), and HBV DNA load were measured at birth and after 7 months. later.Immunoprophylaxis failure was defined as the presence of HBV DNA and HBsAg positivity or the presence of HBV DNA and HBsAg negativity at 7 months. Results: In this prospective cohort study, of the 280 infants enrolled, 14 infants (HBIG/control: 6/8) were lost to follow-up and 266 subjects (HBIG/control: 134/132) completed the 7-month study. The log₁₀HBV DNA load of mothers in the HBIG group and control group were (7.31±0.66) log₁₀IU/ml and (7.32±0.74) log₁₀IU/ml, respectively (P=0.92). The MTCT rate of the two groups was similar (5.97% vs. 7.58%, P=0.63). At 7 months, the HBsAg positive rate and the level of anti-HBs in the two groups were 94.03%(126/134)vs. 91.67% (121/132) and 623.60±412.93 mIU/mL vs. 620.38±399.10 mIU/ml, respectively with no significant difference (P=0.48 and P=0.95, respectively). The log₁₀ HBV DNA load of mothers in immunoprophylaxis failure group and success group was similar (P=0.09). The number of infants who were serum HBsAg positive and HBV DNA positive at birth in the immunoprophylaxis failure group were higher than those in the success group (100% and 100% vs. 35.89% and 31.85%, P<0.01, respectively). The serum HBsAg levels in infants at birth was the only independent relevant factor for HBV MTCT, with risk rates of 11.18 (95% Confidence interval (CI), 1.23-101.88), 352.00 (95%CI, 15.82-7833.20), and 968.00 (95%CI 81.35-11519.19) for HBsAg levels of 0.05-< 1, 1-< 10, and ≥ 10 IU/ml, respectively, compared to infants with HBsAg levels < 0.05 IU/ml. Conclusions Administering 200IU HBIG injection at 1 month did not reduce the risk of HBV MTCT.

OBJECTIVE:To establish a method for the content determination of amygdalin in Lianhua qingwen capsule. METH-ODS:HPLC was performed on the column of Phenomenex Kinetex XB-C18 with mobile phase of acetonitrile-0.2%Phosphoric acid so-lution(6∶94,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 207 nm,column temperature was 30℃,and the injec-tion volume was 10μl. RESULTS:The linear range of amygdalin was 43.16-215.80 μg/ml(r=0.999 7);the limit of detection was 0.431 6μg/ml,the limit of quantitation was 1.294 8μg/ml;RSDs of precision,stability and reproducibility tests no more than 0.69%;recovery was 95.16%-100.49%(RSD=1.67%,n=9). CONCLUSIONS:The method is simple and rapid with high accuracy and well reproducibility,and can be used for the content determination of amygdalin in Lianhua qingwen capsule.