This study investigated the functions and regulatory mechanism of Portulacae Herba and its chemical components on the Toll-like receptor 4(TLR4)/myeloid differentiation primary response 88(MyD88)/nuclear factor kappa B(NF-κB) inflammatory signaling pathway in the colon tissue of mice with dextran sodium sulfate(DSS)-induced ulcerative colitis(UC). A total of 35 mice were randomly divided into groups, including a blank group, a model group, a mesalazine group(0. 5 g·kg~(-1)), and low, medium,and high dose alkaloids from Portulacae Herba groups(9, 18, 36 mg·kg~(-1)), and a combination treatment group, with 5 mice in each group. The blank group was given purified water, while the other groups were continuously given a 3% DSS solution for 7 days to induce the UC model. From day 8 onwards, the treatment group received oral gavage according to the prescribed doses for 14 days. The overall condition, body weight, stool characteristics, and presence of blood in the stool were recorded daily. After the experiment, the disease activity index(DAI) was assessed for each group, and colon length was measured. Histopathological changes in colon tissue were examined using hematoxylin-eosin(HE) staining. The levels of pro-inflammatory cytokines, tumor necrosis factor-α(TNF-α),and interleukin-1β( IL-1β) in serum were measured by enzyme-linked immunosorbent assay( ELISA). The protein and m RNA expression of TLR4, MyD88, and NF-κB in colon tissue were measured using Western blot and quantitative real-time PCR(qPCR).Compared to the blank group, the model group showed a significant decrease in body weight, a notable increase in DAI scores, a significant shortening of colon length, and evident histopathological damage. The levels of inflammatory cytokines TNF-α and IL-1β in the serum were significantly elevated, and the protein and m RNA expression of TLR4, MyD88, and NF-κB in colon tissue were significantly up-regulated. In contrast, the alkaloids from Portulacae Herba treatment groups significantly improved symptoms and reduced body weight loss in mice, decreased DAI scores, alleviated colon shortening, lowered serum levels of TNF-α and IL-1β,significantly down-regulated the expression levels of TLR4, MyD88, and NF-κB proteins and genes in colon tissue, as well as reduced histopathological damage. Therefore, the study suggests that alkaloids from Portulacae Herba can alleviate intestinal inflammation damage in DSS-induced UC mice, with its mechanism involving the TLR4/MyD88/NF-κB signaling pathway.
Animals ; Colitis, Ulcerative/immunology* ; Toll-Like Receptor 4/immunology* ; Myeloid Differentiation Factor 88/metabolism* ; Mice ; NF-kappa B/metabolism* ; Signal Transduction/drug effects* ; Male ; Alkaloids/administration & dosage* ; Drugs, Chinese Herbal/administration & dosage* ; Humans ; Female ; Colon/metabolism* ; Disease Models, Animal

A new method was developed for simultaneous detection of total 19 kinds of organophosphate esters(OPEs)and their diester metabolites(di-OPEs)in human serum(1.0 mL)and urine(1.5 mL)with low volume of samples.The target compounds were determined using ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)after acetonitrile liquid-liquid extraction combined with purification using an ENVI-18 solid-phase extraction(SPE)column.OPEs and di-OPEs were separated using a Shim-pack GIST C18 column(100 mm×2.1 mm,2 μm)with a Shim-pack GIST-HP(G)C18 guard column.An electrospray ionization source(ESI)was employed in mass spectrometry analysis,with positive/negative ion mode using the multiple reaction monitoring(MRM).All target compounds were separated within 15 min,and exhibited good linear relationships in the concentration range of 2-100 ng/mL,with correlation coefficients(R2)above 0.994.The method detection limits(MDL)in serum ranged from 0.001 to 0.178 ng/mL and the MDL in urine ranged from 0.001 to 0.119 ng/mL.The recoveries of the analytes spiked in serum and urine matrices at two concentration levels were 30.5%-126.8%,with the relative standard deviations(RSDs)ranged from 1%to 23%.In addition,paired serum and urine samples from 11 patients were analyzed.For all samples tested,the internal standards of OPEs exhibited recoveries between 61%and 114%,whereas the internal standards for di-OPEs had recoveries ranging from 43%to 103%.OPEs and di-OPEs exhibited high detection frequencies in 22 serum and urine samples.Triethyl phosphate(TEP),tributyl phosphate(TBP),tris(2-ethylhexyl)phosphate(TEHP),tris(2-butoxyethyl)phosphate(TBEP),tris(1-chloro-2-propyl)phosphate(TCIPP),triphenyl phosphate(TPHP),tri-m-tolyl-phosphate(TMTP)and 2-ethylhexyl diphenyl phosphate(EHDPP)were universally detected in all serum samples.TCIPP was identified at the highest concentrations(median 0.548 ng/mL)in serum samples.In urine samples,the detection frequency for 12 kinds of target compounds reached 100%.Notably,TBP emerged as the predominant OPE in urine,demonstrating a median concentration of 0.506 ng/mL.Regarding di-OPEs,bis(2-chloroethyl)phosphate(BCEP)and bis(2-butoxyethyl)hydrogen phosphate(BBOEP)were the most abundant in urine,with median concentrations of 6.404 and 2.136 ng/mL,respectively.The total concentrations of OPEs and di-OPEs in serum and urine were 1.580-3.843 ng/mL and 5.149-17.537 ng/mL,respectively.These results not only confirmed the effectiveness of the method in detection of OPEs and di-OPEs in biological matrices,but also revealed the widespread presence of OPE compounds in human body and pointed to potential exposure risks.

AIM: To investigate the impact of meteorological factors in different environments in the eastern and western regions of China on the incidence of lipid-abnormal dry eye.METHODS: This is a multicenter retrospective study. From March 1, 2021 to February 28, 2022, all patients with dry eye were selected from the ophthalmology clinic of Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine and Friendship Hospital of Xinjiang Yili Kazakh Autonomous Prefecture. General data(gender, age, employment and education)and climate data for outdoor environments(temperature, humidity, air quality index and wind)on the day of the visit were collected. Patients who met the inclusion and exclusion criteria were selected. Single factor, multiple factors and nonlinear model analysis were applied to identify environmental factors of lipid-abnormal dry eye in both regions.RESULTS: There was no significant difference in the incidence of lipid-abnormal dry eye between Nanjing and Yili in different seasons. The incidence of lipid-abnormal dry eye in Yili was significantly higher in all seasons than in Nanjing(P<0.001). The results of univariate research showed that the factors affecting the incidence of lipid-abnormal dry eye were gender, employment, humidity, air quality, and wind. The results of multivariate Logistic regression analysis showed that humidity, temperature and air quality were statistically significant, and remained significant after adjusting for the three confounding factors of age, gender and employment situation. Nonlinear analysis showed that the probability of lipid-abnormal dry eye increased with the decrease of temperature when the temperature was below 10℃; within the range of 10℃～15℃, the probability of lipid-abnormal dry eye tended to stabilize. When the temperature exceeded 15℃, the probability of lipid-abnormal dry eye increased with the increase of temperature. Humidity was negatively correlated with the onset of lipid-abnormal dry eye. As humidity increased, the probability of lipid-abnormal dry eye decreased.CONCLUSIONS: The risk of lipid-abnormal dry eye in Yili is higher than that in Nanjing throughout the four seasons. Humidity, temperature, air quality and other environmental and meteorological factors can all affect the incidence of lipid-abnormal dry eye.

Humans ; Temperature ; Cities/epidemiology* ; Hot Temperature ; Cardiovascular Diseases/epidemiology* ; China/epidemiology* ; Mortality

Abstract: Objective　This article summarizes the clinical characteristics and diagnosis and treatment experience of an elderly patient infected with Omicron variant BA.5.1.3 of COVID-19 in Hainan Province. Methods　The clinical data and treatment of an elderly patient infected with Omicron variant BA.5.1.3 of COVID-19 admitted to Haikou designated hospital on August 15, 2022 were retrospectively analyzed. Results　A 107-year-old female patient was admitted to the hospital with "fever and cough for 1 day". Two of her family members have infected with COVID-19. The patient initially developed fever, accompanied by cough, expectoration, a little white sticky sputum, accompanied by sore throat, muscle pain, fatigue. Nucleic acid test was positive in throat swab, indicating Omicron variant BA.5.1.3 infection. The patient was diagnosed as mild COVID-19 and treated with antiviral therapy, Chinese medicine conditioning, anticoagulation, electrolyte disorder regulation and symptomatic treatment for 9 days. The patient's clinical symptoms were relieved, and she was cured and discharged after two negative nucleic acid tests. One week later, the patient recovered well. Conclusions　Omicron variant BA.5.1.3 is highly infectious, and comprehensive treatment such as antiviral treatment and traditional Chinese medicine treatment has achieved good efficacy. For elderly patients, attention should be paid to maintaining the stability of organ function and internal environment, which is helpful to improve the prognosis of patients.

Arthropods of medical importance such as mosquitoes, ticks and sandflies are one of the key drivers of arthropod-borne diseases outbreak, posing a great threat to global public health security. For further understanding the transmission mechanisms of arthropod-borne diseases and establishing the prevention and control measures, a series of experiments of arthropods infection need to be carried out under laboratory conditions. Besides the regular biosafety requirements, some specific considerations need to be taken into account when performing arthropod infection and the infected arthropod rearing. Except for the physical containment composed of biosafety facilities, a comprehensive assessment of the biosafety risks during operations and corresponding preventive measures are also critical to eliminate or mitigate the biosafety risks. In this paper, we introduce our practice in handling mosquito infection with Risk Group 2 pathogens in Arthropod Containment Level-2 (ACL-2) laboratory, with an aim to provide a reference for researchers in related fields.

Objective: To establish an ultrahigh performance liquid chromatography tandem mass spectrometry method for the determination of creatinine (Cre) and 2-thiothiazolidine-4-carboxylic acid (TTCA) in urine. Methods: In October 2020, the end-of-shift urine samples of the monitored subjects were taken, and the filtrate was prepared by centrifugation. After separated by ultra high performance liquid chromatography C18 column, acetonitrile and 0.2% acetic acid aqueous solution were used as mobile phases for gradient elution, the three quadrupole tandem mass spectrometry adopted an electrospray ion source (ESI) , the ion source temperature was 500 ℃ , and the air curtain gas flow rate was 31.4 L/min, qualitative and quantitative analysis of Cre and TTCA were carried out under the multiple reaction monitoring mode. Results: The linear range of Cre was 1.0-1 000.0 μg/L, the linear equation was y=947.3x-1605.6, and the correlation coefficient was 0.9994. The detection limit and the limit of quantitation were 0.3, 1.0 μg/L. When the addition concentrations were 50.0, 150.0 and 450.0 μg/L, the recovery rates were 92.8%-94.6% , the intra assay precisions were 3.6%-5.7% , and the inter assay precisions were 3.4%-5.4%. The linear range of TTCA was 0.1-200.0 μg/L, the linear equation was y=1164.7x-2243.9, and the correlation coefficient was 0.9991. The detection limit and the limit of quantitation were 0.03, 0.1 μg/L. When the addition concentrations were 10.0, 40.0 and 160.0 μg/L, the recovery rates were 90.8%-93.6%, the intra assay precisions were 4.6%-7.4%, and the inter assay precisions were 4.4%-6.9%. Conclusion: The sample pretreatment process of the ultra high performance liquid chromatography tandem mass spectrometry method for the determination of Cre and TTCA in urine is simple, and the continuous determination of Cre and TTCA in urine can be realized only by switching mass spectrometry parameters under the same chromatographic conditions, which is accurate and efficient, and each performance index of the method meets the determination requirements.
Chromatography, High Pressure Liquid ; Creatinine ; Humans ; Tandem Mass Spectrometry ; Thiazolidines

Objective: To explore the correlation of ultrasound-guided fine-needle aspiration(US-FNA) combined with BRAF V600E mutation detection and ultrasound features and central cervical lymph nodes metastasis of classic papillary thyroid cancer(PTC) for providing a reliable molecular basis for clinical preoperative evaluation of patients. Methods: Ninty-three cases of patients collected from October 2017 to November 2018 in Gansu Province Hospital were enrolled, who underwent general ultrasonic examination TI-RADS ≥4a, the US-FNA highly suspicious of PTC, thyroid surgery including total thyroidectomy and central cervical lymph node dissection, with the postoperative pathologic results of classical PTC and whether the central cervical lymph node metastasis happened in the patients. Part of the specimen applied HE staining for cytological diagnosis, the other part of specimen was used real-time for detection of BRAF V600E gene mutation by fluorescent quantitative polymerase chain reaction (PCR) method. Results: Univariate analysis showed that the occurrence of cervical lymph node metastasis for classic PTC were significantly correlated with gender(χ²=10.303, P=0.002), BRAF V600E mutation(χ²=31.204, P=0.000) and extrathyroidal invasion(χ²=12.848, P=0.000). Multi-logistic regression analysis showed that BRAF V600E mutation(OR=13.324, 95%CI=4.058-43.744, P=0.000) and extrathyroidal invasion(OR=5.738, 95%CI=1.766-18.643, P=0.004) were the risk predictors of cervical lymph node metastasis of classic PTC. Gender(OR=0.385, 95%CI=0.112-1.324, P=0.130) was not the risk predictor. Conclusions US-FNA combined with BRAF V600E mutation and extrathyroidal invasion are the risk factors in predicting central cervical lymph node metastasis in classic PTC. Patients with these two risk factors should be elected to undergo prophylactic central cervical lymph node dissection.

Hispidulin(HPDL) chitosan microspheres were prepared in this study to deliver HPDL to the lesion sitevia intravenous injection, and further evaluate their anticancer effects in vitro and the growth inhibition effect on A549 cells spheroids. HPDL chitosan microspheres were prepared by emulsion crosslinking method with chitosan as a drug carrier and the amount of HPDL was determined by high performance liquid chromatography (HPLC). The morphology of microspheres was observed under laser scanning confocal microscope. Additionally, the drug release amount of targeting microspheres was detected by dialysis method. Furthermore, the anti-proliferative effects against A549 lung cancer cells were tested by sulforhodamine B (SRB) method, and the effects of HPDL chitosan micrpsphereson early apoptosis of A549 cellswere determined by flow cytometry. A549 cells tumor spheroids were developed in vitro and then HPDL chitosan microspheres were added. On the 0, 1, 3, 7 d after adding the drugs, the inverted microscope was used to observe the mythologicaland volume changes of A549 cells spheroids. The encapsulation efficiency of HPDL chitosan microspheres was （75.32±0.52）%, and the drug loading amount was （7.76±0.67）%. Meanwhile, the microspheres were round shaped andhad smooth surface. The HPDL chitosan microspheres exhibited stronger inhibitory effects on A549 lung cancer cells. The results of flow cytometry indicated that, the early apoptosis rate of lung cancer A549 cells was （37.0±0.75）% at 24 h cells culture after drug administration. The volume of tumor spheroid was significantly inhibited, which had been shrunk by (50.09±11.06)% after the treatment by drug-loaded microsphere at day 7 as compared with blank group; meanwhile, the cells surface were obviously lysed. The preparation method in this research was simple and practicable, and the microspheres prepared with this method were round and smooth, with high encapsulation efficiency, which can significantly inhibit proliferation of lung adenocarcinoma A549 cells and induce cell apoptosis, and at the same time can cause lysisand death of A549 cell tumor spheroid.

Objective To analyze the changes of varicella outbreak in Minhang District before and after practicing "the varicella emergency vaccination implementation program (2013-2014 trial version),Shanghai",and to evaluate the effects of live attenuated vaccine against varicella.Methods We used cross-sectional investigation to collect monitoring data of vaticella outbreak recorded in "China Information System for Disease Control and Prevention".The observation periods before and after the implementation of the program were during 2012-01-01 to 2013-06-30 and during 2013-07-01 to 2014-12-31.Results Before the implementation of the program,14 cases of varicella epidemic were reported in Minhang District.All cases were up to the outbreaks in the epidemic,the longest duration was 82 days,the average duration continued was 50.86 days (912/14),the average cumulative generation was 2.42 generations (50.86/21),the total number of people was 159,the total incidence rate was 1.26％ (159/12 625),the class incidence rate was 9.09％ (159/1 750),the average number of people was 11.36 (159/14),the average number of class was 3.21 (45/14).After the implementation of the program,169 cases of the varicella epidemic were reported in Minhang District,only 22 cases rose to the outbreaks in the epidemic,the longest duration was 66 days,the average duration continued was 41.32 days (909/22),the average cumulative generation was 1.97 generations (41.32/21),the total number of people was 200,the total incidence rate was 1.10％ (200/18 200),the class incidence rate was 8.38％ (200/2 387),the average number of people was 9.09 (200/22),the average number of class was 2.50 (55/22);Although the varicella epidemic in the total outbreaks had significantly increased in Minhang District before and after emergency implementation,but the varicella outbreak in the varicella epidemic significantly decreased,the total incidence rate,the class incidence rate,the longest duration,the average duration,the average cumulative generation,the average number of people and the average number of class had decreased in varying degrees.There is statistical difference between the average duration before and after the implementation (x2 =4.06,P＜0.05).Conclusions Emergency vaccination of varicella vaccine in high risk population can effectively control the spread of epidemic situation and reduce the possibility of secondary transmission.