Objective To investigate the effect of the aqueous extract of Chuan Xiong Rhizoma(CR)on brain metastasis of melanoma B16F10 cells in mice.Methods C57BL/6J mouse models of brain metastasis of melanoma were established by ultrasound-guided intraventricular injection of Luc-labeled B16F10 cells,and brain tumor growth was monitored by in vivo imaging.The mouse models were then randomized for daily gavage of saline or aqueous extract of CR(equivalent crude drug concentration of 1 mg/g).Behavioral tests were used to evaluate the neuroprotective effects of CR in the tumor-bearing mice,and the changes in proteins associated with blood-brain barrier integrity,neuronal cell proliferation and apoptosis,and microglial cell apoptosis and activation were observed using immunofluorescence assay.The efficacy of CR combined with temozolomide(25 mg/kg)against brain metastases of B16F10 cells was observed by in vivo imaging.Results CR-treated mouse models did not show obvious progression of brain metastases and had a reduced rate of body weight loss and lowered protein expressions of ZO-1,claudin-5,occludin,P-gp,TNF-α,AQP4 and PDGFRβ.In the behavioral tests,the CR-treated mice showed prolonged stay on the wooden stick with a shortened time of sticky stick removal.Immunofluorescence assay showed increased proliferation and decreased apoptosis of neuronal cells and microglia in CR-treated mice.CR treatment significantly increased the levels of CD86,CD206,IL-4 and IL-10 and decreased the levels of CD163 and IL-1β in the microenvironment of brain metastases.The mice receiving combined treatments with CR and temozolomide showed significantly lower intensity of fluorescent signals in the brain than those treated with temozolomide alone.Conclusion CR does not promote brain metastasis of melanoma while inducing opening of the blood-brain barrier,and its combined use with TMZ results in enhanced inhibition against brain metastasis of melanoma B16F10 cells in mice.

Objective To investigate the effect of the aqueous extract of Chuan Xiong Rhizoma(CR)on brain metastasis of melanoma B16F10 cells in mice.Methods C57BL/6J mouse models of brain metastasis of melanoma were established by ultrasound-guided intraventricular injection of Luc-labeled B16F10 cells,and brain tumor growth was monitored by in vivo imaging.The mouse models were then randomized for daily gavage of saline or aqueous extract of CR(equivalent crude drug concentration of 1 mg/g).Behavioral tests were used to evaluate the neuroprotective effects of CR in the tumor-bearing mice,and the changes in proteins associated with blood-brain barrier integrity,neuronal cell proliferation and apoptosis,and microglial cell apoptosis and activation were observed using immunofluorescence assay.The efficacy of CR combined with temozolomide(25 mg/kg)against brain metastases of B16F10 cells was observed by in vivo imaging.Results CR-treated mouse models did not show obvious progression of brain metastases and had a reduced rate of body weight loss and lowered protein expressions of ZO-1,claudin-5,occludin,P-gp,TNF-α,AQP4 and PDGFRβ.In the behavioral tests,the CR-treated mice showed prolonged stay on the wooden stick with a shortened time of sticky stick removal.Immunofluorescence assay showed increased proliferation and decreased apoptosis of neuronal cells and microglia in CR-treated mice.CR treatment significantly increased the levels of CD86,CD206,IL-4 and IL-10 and decreased the levels of CD163 and IL-1β in the microenvironment of brain metastases.The mice receiving combined treatments with CR and temozolomide showed significantly lower intensity of fluorescent signals in the brain than those treated with temozolomide alone.Conclusion CR does not promote brain metastasis of melanoma while inducing opening of the blood-brain barrier,and its combined use with TMZ results in enhanced inhibition against brain metastasis of melanoma B16F10 cells in mice.

Objective:To investigate the prognostic value of cardiac ultrasound left ventricular ejection fraction (LVEF) on admission in patients with septic cardiomyopathy.Methods:A retrospective cohort study was conducted. The patients with septic cardiomyopathy hospitalized in the intensive care unit of Zhoupu Hospital Affiliated to Shanghai Health College from January 2019 to March 2023 were enrolled. The general information including gender and age, LVEF on admission, severity of illness scores within 24 hours after admission [acute physiology and chronic health evaluationⅡ (APACHEⅡ) score and sequential organ failure assessment (SOFA) score], procalcitonin (PCT), cardiac biomarkers [N-terminal pro-brain natriuretic peptide (NT-proBNP), cardiac troponin T (cTnT), and MB isoenzyme of creatine kinase (CK-MB)], mitochondria related indicators [aspartate aminotransferase (AST), AST/alanine aminotransferase (ALT) ratio], blood lactate (Lac), the usage of vasoactive drugs and mechanical ventilation, and the prognosis during hospitalization were collected. The differences in above clinical data between the two groups were compared. The variables with statistically significant differences in univariate analysis were incorporated into multivariate Logistic regression analysis to analyze the independent risk factors for death during hospitalization in patients with septic cardiomyopathy. Receiver operator characteristic curve (ROC curve) was drawn to evaluate the prognostic value of LVEF by echocardiography on admission in patients with septic cardiomyopathy during hospitalization.Results:A total of 62 patients were enrolled, including 36 males and 26 females. Thirty-nine cases died and 23 cases survived during hospitalization, and the mortality was 62.90%. Compared with the survival group, the LVEF of patients on admission was lower in the death group [0.51 (0.40, 0.57) vs. 0.56 (0.51, 0.63), P < 0.01], APACHEⅡ score, SOFA score, Lac, NT-proBNP, CK-MB within 24 hours after admission were higher [APACHEⅡ score: 22.18±8.38 vs. 17.39±8.22, SOFA score: 9.90±3.87 vs. 7.09±3.27, Lac (mmol/L): 5.10 (2.63, 11.50) vs. 2.00 (1.40, 5.00), NT-proBNP (μg/L): 5.24 (2.84, 11.29) vs. 2.53 (0.35, 6.63), CK-MB (U/L): 1.88 (0.21, 5.33) vs. 0.17 (0.02, 1.62), all P < 0.05], and the proportion of vasoactive drug application was higher (82.05% vs. 47.83%, P < 0.01). Multivariate Logistic regression analysis showed that LVEF on admission was an independent risk factor for predicting the prognosis of patients with septic cardiomyopathy during hospitalization [odds ratio ( OR) = 0.920, 95% confidence interval (95% CI) was 0.855-0.990, P = 0.025]. ROC curve analysis showed that the area under the ROC curve (AUC) of LVEF on admission for predicting the death of patients with septic cardiomyopathy was 0.715 (95% CI was 0.585-0.845, P = 0.005). When LVEF ≤ 0.52, the sensitivity was 73.9%, and the specificity was 61.5%. Conclusions:The lower cardiac ultrasound LVEF on admission, the worse the prognosis of patients with septic cardiomyopathy. The cardiac ultrasound LVEF on admission can be used as a clinical index to evaluate the severity of the condition and predict the prognosis of patients with septic cardiomyopathy.

Objective:To understand the contents of edible salt iodine and urinary iodine of children and pregnant women in Yunnan Province, and to evaluate the iodine nutrition status, so as to provide a basis for scientific prevention and treatment of iodine deficiency disorders (IDD).Methods:From November to December 2020, one county (city) was selected from each prefecture (city), two townships (towns and streets) were selected from each county (city) and two villages (neighborhood committees) were selected from each township (town and street) from each of the 16 prefectures (cities) in Yunnan Province as the investigation sites. A total of 20 non-boarding children (male and female balanced) aged 8 - 10 years old were selected from each primary school in each village (neighborhood committee) to collect salt and urine samples. A total of 80 children were investigated in each county (city). A total of 20 pregnant women were selected from each township (town and street) to collect salt and urine samples. A total of 40 pregnant women were investigated in each county (city). All salt samples and urine samples were tested for iodine contents.Results:A total of 2 009 salt samples and 2 041 urine samples (1 375 for children, 666 for pregnant women) were collected from children aged 8 - 10 years old and pregnant women in 16 counties (cities) of Yunnan Province. Among them, the median salt iodine was 26.0 mg/kg, the coverage rate of iodized salt was 100.0% (2 009/2 009), the qualified rate of iodized salt was 98.7% (1 982/2 009), and the consumption rate of qualified iodized salt was 98.7% (1 982/2 009). The difference of salt iodine content in key populations in different counties (cities) was statistically significant ( H = 258.98, P < 0.01). The median urinary iodine of children aged 8 - 10 years old was 188.5 μg/L. There was statistically significant difference in urinary iodine content among children of different ages ( H = 29.45, P < 0.01), but there was no statistically significant difference in urinary iodine content among children of different genders ( H = 1.43, P > 0.05). In addition, the median urinary iodine of pregnant women was 141.9 μg/L, 52.1% (347/666) was < 150 μg/L. There was statistically significant difference in urinary iodine content of pregnant women in different counties (cities, H = 88.32, P < 0.01). Conclusions:The qualified rate of iodized salt, the coverage rate of iodized salt and the consumption rate of qualified iodized salt in key populations of Yunnan Province are more than 90%, and the iodized salt supply is good. Iodine nutrition of children aged 8 - 10 years old is at an appropriate level (100 - 199 μg/L); iodine nutrition of pregnant women is in an state of iodine deficiency ( < 150 μg/L). It is suggested to strengthen IDD monitoring and health education among key populations, improve residents' awareness of disease prevention, and make scientific iodine supplementation.

Objective:To evaluate the prognostic value of Karnofsky performance scores (KPS) in elderly patients with sepsis, so as provide a basis for clinical evaluation of the condition, prognosis and corresponding treatment measures.Methods:A retrospective cohort study was conducted to collect the general information, clinical data, and follow-up data of limb motor function status and self-care ability of elderly patients with sepsis who were hospitalized in the Intensive Care Unit of our hospital from January 2018 to June 2021. Patients were divided into the survival group and death group according to whether they survived the hospitalization. Statistical analysis was performed using t-test, chi-square test, and Mann-Whitney test. The KPS score before admission, disease severity scores (APACHEⅡ and SOFA), serum procalcitonin (PCT), N-terminal pro-brain natriuretic peptide (NT-proBNP), and arterial blood lactate level on admission were compared between the two groups. Then, the factors with significance in univariate analysis were analyzed by Logistic regression method, the independent risk factors for predicting in-hospital mortality were determined, and the receiver operating characteristic (ROC) curve was drawn to evaluate the prognostic value of KPS score in elderly patients with sepsis.Results:A total of 135 patients were collected. There were 85 males and 50 females, 60 died and 75 survived during hospitalization, with a mortality rate of 44.4%. The preadmission KPS score of elderly patients with sepsis in the death group was significantly lower than that in the survival group [30 (30, 40) vs. 70 (50, 90), P<0.001]. Multivariate logistic regression analysis showed that KPS score ( OR=0.938, 95% CI: 0.914-0.963, P<0.001), SOFA score ( OR=1.255, 95% CI: 1.066-1.451, P=0.002) and arterial blood lactate ( OR=1.219, 95% CI: 1.059-1.404, P=0.006) were independent risk factors for predicting the prognosis during hospitalization. ROC curve analysis of mortality showed that compared with SOFA score and blood lactate, the area under the curve of KPS score was the largest, with AUC of 0.830 (95% CI: 0.756-0.890, P<0.001). In addition, the combination of KPS, SOFA and blood lactate had a greater predictive value for the prognosis of elderly patients with sepsis than that of the single index, with an AUC of 0.883 (95% CI: 0.826-0.940, P<0.001). Conclusions:The lower the KPS score, the worse the prognosis of elderly patients with sepsis. The KPS score can be used as a clinical indicator to predict the prognosis of elderly patients with sepsis.

This paper aims to detect the antigens in porcine circovirus type 2 (PCV2) vaccines by high-performance size-exclusion chromatography (HPSEC) coupled with multi-angle laser light scattering (MALLS). With purified inactivated PCV2 and PCV2 virus-like particles (VLP) as references, two inactivated vaccines (a and b) and two VLP vaccines (c and d) for PCV2 from four manufacturers were analyzed by HPSEC-MALLS after demulsification. The antigen peaks in HPSEC-MALLS were identified by PCV2 antigen test strips, Western blotting and transmission electron microscope (TEM). The repeatability and linearity of the method were investigated. The results showed the virus antigens in the two inactivated vaccines were eluted at about 13.3 min in HPSEC. The molecular weight of these antigens was 2.61×10⁶ (±4.34%) Da and 2.40×10⁶ (±2.51%) Da, respectively, as calculated by MALLS. The antigen peaks of the two VLP vaccines also appeared at 13.3 min and the molecular weight was 2.09×10⁶ (±2.94%) Da and 2.88×10⁶ (±11.85%) Da, respectively, which was close to the theoretical molecular weight of PCV2. Moreover, an antigen peak of VLP vaccine c was observed at 11.4 min and the molecular weight was 4.37×10⁶ (±0.42%) Da. The antigen was verified to be the dimer of VLP by TEM. Vaccine d and purified Cap VLP antigens were tested repeatedly, and the RSD of the peak area (n=3) was all < 1.5%, indicating that the method was repeatable. The purified VLP were diluted in serial and tested for linearity. The result suggested good linear relationship between the peak area of VLP or VLP aggregates and the protein concentration of the sample with R² of 0.999 and 0.997, respectively. Thus, the method met the requirement for quantification and aggregate analysis. This method is accurate and efficient in in vitro quality evaluation and improvement of PCV2 vaccine.
Animals ; Antibodies, Viral ; Capsid Proteins ; Chromatography, Gel ; Circoviridae Infections/prevention & control* ; Circovirus ; Lasers ; Swine ; Vaccines, Inactivated ; Vaccines, Virus-Like Particle ; Viral Vaccines

Objective:To characterize limb traumatic osteomyelitis in a regional trauma center in South China.Methods:The case system at Division of Orthopaedics and Traumatology, Department of Orthopaedics, Nanfang Hospital was searched for the data of confirmed limb traumatic osteomyelitis from January 1, 2010 through September 1, 2019. The clinical items collected were gender, age of onset, cause of injury, injury nature (open or closed), infected site (single site or multiple sites), intraoperative culture of pathogenic microorganisms, number and type of pathogenic microorganisms, and amputation. The above data were sorted out and analyzed statistically.Results:A total of 674 patients suffered limb traumatic osteomyelitis in South China with a male to female ratio of 4.81∶ 1. The limb traumatic osteomyelitis resulted mainly from open injury (64.09%, 432/674). It was mainly caused by a traffic accident (39.51%, 211/534). It was featured by single-site infection (83.68%, 564/674). It affected mainly the lower limbs: the tibia (53.19%, 300/564), the femur (18.97%, 107/564), the calcaneus (11.70%, 66/564) and other foot bones (4.61%, 26/564). The positive rate of intraoperative microorganisms culture was 65.26% (355/544). In the majority of patients (74.65%, 265/355), the traumatic osteomyelitis was caused by infection of a single pathogenic microorganism. The most common single pathogenic microorganism was Staphylococcus aureus (38.11%, 101/265), followed by Pseudomonas aeruginosa (20.00%, 53/265). The rate of amputation related to traumatic osteomyelitis was 3.71% (25/674), and the overall disability rate was 4.45% (30/674).Conclusions:In South China, more males were prone to limb traumatic osteomyelitis. Most cases were secondary to an open fracture and caused by a traffic accident. Traumatic osteomyelitis usually occurred at a lower limb. The most common pathogenic microorganism was Staphylococcus aureus.

Early management of open extremity fractures and treatment of chronic osteomyelitis,in general,require thorough debridement.Radical debridement of the contaminated or infected bone tissues usually results in bone defects.If such bone defects are not dealt with properly,they can easily deteriorate into infected ones.Treatment of infected bone defects can be extremely complicated.Application of local antibiotic delivery system can simultaneously fill the bone defects,prevent and treat the infection by delivering a high concentration of antibiotics to the lesion.The clinical antibiotic carriers can be divided roughly into 2 types:non-biodegradable and biodegradable.The biodegradable type involves bioceramic,natural polymer,synthetic polymer and composite materials.This literature review summaries the characteristics and clinical applications of current local antibiotic carriers.

We developed a pre-treatment method to remove interfering substances during quantification of 146S antigens in foot-and-mouth disease (FMD) vaccines by high performance size exclusion chromatography (HPSEC). Three methods, including ultracentrifugation, PEG precipitation and nuclease digestion, were optimized and compared for removal efficiency of the interfering impurities in FMD vaccines. Under optimized conditions, the 146S contents in two batches of FMD vaccines were determined to be 7.1 and 7.6 μg/mL by ultracentrifugation, 9.7 and 10.4 μg/mL by PEG precipitation, and 10.5 and 10.4 μg/mL by nuclease digestion. The optimal condition for nuclease digestion using Benzonase determined by response surface method was as follows: appending Benzonase into 200 μL of antigen phase to a final concentration of 421 U/mL and incubating at 25.1 °C for 1.29 h. This method has advantages including efficient removal of the interfering impurities, fast processing speed, and mild operating conditions. Then 12 bathes of FMD vaccines with different serotypes produced by 4 manufacturers were tested to verify the established treatment method. Results showed the method was applicable to various FMD vaccines with good reproducibility (RSD<5.3%, n=3). The developed method removed interference from impurities during quantification of 146S, and therefore would broaden the application of HPSEC in vaccine quality control and ensure the accuracy and reliability.
Animals ; Chromatography, Gel ; Foot-and-Mouth Disease ; Foot-and-Mouth Disease Virus ; Reproducibility of Results ; Viral Vaccines

The aim of this study is to quantify the 146S antigen in foot-and-mouth disease virus (FMDV) inactivated vaccine by size-exclusion chromatography (SEC). The analysis was performed on a TSKgel G4000SWXL column (7.8 mm×30 cm), with a pH 7.2 buffer salt system as the mobile phase. The flow rate was 0.6 mL/min, the injection volume was 100 μL and the detection wavelength was 259 nm. The calibration curve was established by using purified inactivated FMDV (serotype O) 146S antigen; 3 batches of vaccine formulated by inactivated antigen solution were tested to verify the accuracy, reproducibility, specificity and tolerability of the method. At last 16 batches of vaccine were determined by the SEC method. Results showed a good linearity between peak area and concentration of 146S antigen in the range between 0.56 and 67.42 μg/mL (R2=0.996, n=10), and the average recovery rate of 146S antigen in the 3 batches of vaccine formulated in lab were 93.6% (RSD=2.7%, n=3), 102.3% (RSD=2.6%, n=3), and 95.5% (RSD=5.1%, n=3). The method was proved accurate and reliable with good reproducibility (RSD=0.5%, n=6), and applied to determine 16 batches of the commercial FMDV vaccine. According to the above results, the SEC method is high effective for 146S antigen quantify in the inactivated FMDV vaccine and would provide strong support for the vaccine quality control.