Objective To develop an ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method to simultaneously determine 10 main components, including berberine, phellodendrine, specnuezhenide, mangiferin, loganin, paeoniflorin, geniposide, baicalin, and acteoside in Compound Dihuang oral solution. Methods An UPLC-MS/MS method was established with an ACQUITY UPLC BEH-C18 （2.1 mm×100 mm, 1.7 μm）column and mobile phase of 0.1% formic water（A）-methanol solution（B） in a gradient elution manner. The flow rate of mobile phase was 0.2 ml/min.The temperature of column was 30℃. The injection volume was 2 μl. The MS detection was in MRM mode. Results 10 components in Compound Dihuang oral solution had a good linear relationship within their concentration range,and the precision, repeatability, stability and recovery met the requirements. The contents of berberine, phellodendrine, specnuezhenide, mangiferin, loganin, paeoniflorin, geniposide, baicalin, and acteoside in 7 batches of samples were （89.7-95.6） μg/ml, （164.0-177.7） μg/ml, （540.0-610.0） μg/ml, （408.7-429.0） μg/ml, （726.0-825.0） μg/ml, （503.7-572.0） μg/ml, （1380.0-1540.0） μg/ml, （2596.7-2896.7) μg/ml, （4866.7-5520.0) μg/ml, （61.8-67.2) μg/ml, respectively. Conclusion The established method was easy to be repeated and operated. The contents of 10 components in Compound Dihuang oral solution could be determined quickly and accurately, which provided a reference for the quality control of hospital preparation.

Humans ; Aged ; Lung Diseases ; Pneumonia/drug therapy* ; Adrenal Cortex Hormones/therapeutic use* ; Pulmonary Disease, Chronic Obstructive/drug therapy* ; Administration, Inhalation ; Community-Acquired Infections/drug therapy*

ObjectiveTo explore the common syndrome elements of cerebral ischemic stroke （CIS） complicated with obstructive sleep apnea-hypopnea syndrome （OSAHS）， reveal the characteristics of traditional Chinese medicine （TCM） syndromes of the disease， clarify the syndrome differentiation and syndrome types， provide evidence for clinical syndrome differentiation， and provide reference for establishing the TCM syndrome type standards of CIS complicated with OSAHS. MethodThe clinical information form of CIS complicated with OSAHS formulated by the research group was used for syndrome survey， and the four-examination information of 300 patients with CIS complicated with OSAHS was collected. The four-examination information of patients was analyzed by latent structure method and comprehensive cluster analysis， and the common syndrome elements of CIS complicated with OSAHS were extracted by combining the TCM basic theory and clinical experience. On this basis， the characteristics of TCM syndromes and the syndrome types in line with reality were summarized. ResultThe top five syndrome elements in patients with CIS and OSAHS are sleep snoring， open mouth breathing， physical obesity， night awakening and dizziness. The top five tongue and pulse manifestations are enlarged tongue， slippery pulse， slippery coating， thick and white coating and purple tongue. The disease locations are the lung， spleen， stomach， kidney， liver and brain. The nature of disease includes deficiency， depression， blood stasis， phlegm， dampness and fire. The clinical syndrome types include the syndrome of stagnation of phlegm and dampness， syndrome of phlegm-dampness blocking the mind， syndrome of spleen deficiency with dampness， syndrome of Yin deficiency leading to fire hyperactivity， syndrome of Qi depression blocking collaterals， syndrome of liver depression and blood stasis， syndrome of Qi deficiency with dampness， and syndrome of Yang deficiency induced water retention. ConclusionIn addition to the common phlegm-， dampness- and blood stasis-related syndromes in patients with CIS and OSAHS， there are also depression- and deficiency-related syndromes. The main etiology and pathogenesis is the disturbance of Qi movement. In clinical practice， attention should be paid to the specific situation of individual patients to differentiate between deficiency and excess， and the treatment should be performed by the method of soothing and reinforcing， or unblocking and clearing， or both.

ObjectiveTo observe the effects of Jianpi Bushen Huoxue prescription （JPBSHX） on rat brain microvascular endothelial cells （RBMECs） based on hypoxia-inducible factor-1α （HIF-1α）/vascular endothelial growth factor （VEGF） signaling pathway， aiming to provide a theoretical basis for the treatment of ischemic stroke. MethodTwelve 8-week-old male SPF-grade SD rats were selected. Eight of them were randomly chosen and given 3.25 g·mL^-1 JPBSHX solution by gavage at a dose of 10 mL·kg^-1 for 5 consecutive days to prepare the medicated serum， which was then preserved for later use. The remaining four rats were given the same volume of normal saline. Follow-up operations were the same as those of the above eight rats. Normal rat serum was collected and stored for later use. RBMECs were revived， cultured， passaged， and randomly divided into five groups： normal group （20% normal rat serum+80% high glucose DMEM）， model group （hypoxia-reoxygenation injury） （20% normal rat serum+80% glucose-free DMEM）， medicated serum group （20% JPBSHX-medicated serum+80% glucose-free DMEM）， medicated serum+HIF-1α inhibitor group （20% JPBSHX-medicated serum+HIF-1α inhibitor 1 mg +80% glucose-free DMEM）， and medicated serum+VEGF inhibitor group （20% JPBSHX-medicated serum +VEGF inhibitor 1 mg+80% glucose-free DMEM）. The relative protein expression levels of Claudin-1 and Claudin-5 in RBMECs， the expression levels of HIF-1α and VEGF in RBMEC culture supernatants， the repair ability of RBMECs， and the number of nodes， microvessels， and their lengths after 72 h of culture were observed in each group. ResultAfter 24 h of reoxygenation， the scratch healing rate in the model group was significantly lower than in the normal group （P<0.01）. Compared with the result in the model group， the scratch healing rates significantly improved in the medicated serum group， medicated serum+HIF-1α inhibitor group， and medicated serum+VEGF inhibitor group （P<0.05）. However， the healing rates in the medicated serum+HIF-1α inhibitor group and medicated serum+VEGF inhibitor group were significantly lower than that in the medicated serum group （P<0.05）. The number of nodes， microvessels， and total length of microvessels in the model group were significantly lower than those in the normal group （P<0.01）. These indicators significantly improved in the medicated serum group， medicated serum+HIF-1α inhibitor group， and medicated serum+VEGF inhibitor group compared with those in the model group （P<0.05）， but were significantly lower in the medicated serum+HIF-1α inhibitor group and medicated serum+VEGF inhibitor group compared with those in medicated serum group （P<0.05）. The relative expression levels of Claudin-1 and Claudin-5 proteins were significantly lower in the model group than in the normal group （P<0.01）. These levels were significantly higher in medicated serum group， medicated serum+HIF-1α inhibitor group， and medicated serum+VEGF inhibitor group than those in the model group （P<0.05）， but were significantly lower in the medicated serum+HIF-1α inhibitor group and medicated serum+VEGF inhibitor group than those in the medicated serum group （P<0.05）. The expression levels of HIF-1α and VEGF in the RBMEC culture supernatants were significantly lower in the model group than those in the normal group （P<0.01）. These levels were significantly higher in the medicated serum group， medicated serum+HIF-1α inhibitor group， and medicated serum+VEGF inhibitor group than those in the model group （P<0.05）， but were significantly lower in the medicated serum+HIF-1α inhibitor group and medicated serum+VEGF inhibitor group than those in the medicated serum group （P<0.05）. ConclusionJPBSHX can promote the proliferation， migration， and angiogenesis， such as tubule formation， of RBMECs damaged by hypoxia-reoxygenation injury， and this effect may be achieved through the regulation of the HIF-1α/VEGF signaling pathway.

Objective:To investigate the relationship between plasma fluoride content, daily calcium intake and blood cell parameters in children and adolescents.Methods:This study was based on the National Health and Nutrition Examination Survey (NHANES) database of the United States from 2013 to 2016, with 3 684 children and adolescents aged 6 - 19 as the research subjects. Information on plasma fluoride content, daily calcium intake and blood cell parameters from the database were collected. Non-linear relationships between plasma fluoride content, daily calcium intake and blood cell parameters were analyzed using restricted cubic splines. If there was a non-linear relationship, the optimal inflection point was calculated using threshold/saturation effect analysis method. Subsequently, multiple linear regression models were used to analyze the associations among the three, and the modification effect of daily calcium intake (binary classification, stratified by median daily calcium intake) on the association between plasma fluoride content and blood cell parameters was analyzed.Results:There was no non-linear relationship between plasma fluoride content and white blood cell count, hemoglobin content and platelet count ( Pnon-linear > 0.05), but there was a non-linear relationship between plasma fluoride content and erythrocyte count and hematocrit ( Pnon-linear < 0.001). After adjusting for confounding factors, the optimal inflection points of the effects of plasma fluoride content on erythrocyte count and hematocrit were 0.54 and 0.31 μmol/L, respectively. There was no non-linear relationship between daily calcium intake and blood cell parameters ( Pnon-linear > 0.05). After adjusting for confounding factors, for every 1 μmol/L increase in plasma fluoride content, the white blood cell count increased by 0.49 × 10 9/L ( P = 0.009). There was a saturation effect in the association between plasma fluoride content, erythrocyte count and hematocrit: when plasma fluoride content was < 0.54 μmol/L, the erythrocyte count decreased by 0.46 × 10 12/L for every 1 μmol/L increase ( P < 0.001). When plasma fluoride content was < 0.31 μmol/L, the hematocrit decreased by 6.29% for every 1 μmol/L increase ( P = 0.006). The above associations were not statistically significant when plasma fluoride content was higher than the optimal inflection points ( P > 0.05). After stratification according to the median daily calcium intake, in the low-calcium group (daily calcium intake < 0.87 g), for every 1 μmol/L increase in plasma fluoride content, the white blood cell count increased by 0.77 × 10 9/L ( P = 0.001). When plasma fluoride content was < 0.54 μmol/L, the erythrocyte count decreased by 0.41 × 10 12/L for every 1 μmol/L increase ( P = 0.002). When plasma fluoride content was ≥0.54 μmol/L, erythrocyte count decreased by 0.47 × 10 12/L for every 1 μmol/L increase ( P < 0.001). When the plasma fluoride content was < 0.31 μmol/L, the hematocrit decreased by 8.29% for every 1 μmol/L increase ( P = 0.011). The above associations were not statistically significant in the high-calcium group (daily calcium intake ≥0.87 g, P > 0.05). There was an interaction of daily calcium intake and plasma fluoride content on platelet count ( Pinteraction = 0.070), as demonstrated by an increase in platelet count of 12.68 × 10 9/L ( P = 0.013) in the low-calcium group and a decrease in platelet count of 9.05 × 10 9/L ( P = 0.035) in the high-calcium group for every 1 μmol/L increase in plasma fluoride content. Conclusions:The blood cell parameters of children and adolescents are closely related to plasma fluoride content, but not directly related to daily calcium intake. However, the correlation between plasma fluoride content and blood cell parameters varies among different calcium intake populations, and daily calcium intake can modify the association between plasma fluoride content and platelet count.

Objective To analyze the prognostic value of serum microRNA(miRNA)-21-5p and miRNA-5189-5p expression levels in elderly patients with prostate cancer after laparoscopic radical prostatectomy(LRP).Methods A total of 213 elderly prostate cancer patients treated with LRP from January 2014 to Sep-tember 2018 in the First Affiliated Hospital of Hainan Medical College were retrospectively selected as the study objects.According to the 5-year follow-up outcome,the patients were divided into good prognosis group(87 cases)and poor prognosis group(126 cases).The preoperative clinical baseline data and the expression levels of serum miRNA-21-5p and miRNA-5189-5p were collected.Logistic regression analysis was used to screen the factors affecting the prognosis of patients.The prognostic prediction model was constructed with independent risk factors,and the predictive value was analyzed by drawing receiver operating characteristic(ROC)curve.Results Multivariate Logistic regression analysis showed that serum prostate specific antigen(PSA)level,positive incisal margin,and expression levels of miRNA-21-5p and miRNA-5189-5p were inde-pendent risk factors for the prognosis of elderly prostate cancer patients receiving LRP(P＜0.05).The com-bined prediction model was established based on independent risk factors,and the area under the curve of each factor alone predicting prognosis of elderly prostate cancer patients receiving LRP was less than that of four combined prediction(P＜0.05).Conclusion The expression levels of serum miRNA-21-5p and miRNA-5189-5p are of high value in predicting the prognosis of elderly prostate cancer patients after LRP.

IgA nephropathy is a common primary glomerular disease.Its pathogenesis is currently unclear.As an autoimmune disease, immune response is a key link in the onset of IgA nephropathy.Multiple immune cells are involved in the occurrence and development of IgA nephropathy.The impacts of different cell subpopulations of innate and adaptive immunity can lead to sustained immune damage directly or indirectly.This article elaborates on the role of immune cells in IgA nephropathy from two aspects, namely innate immunity and adaptive immunity, aiming to provide references for further research on IgA nephropathy.

Objective To investigate the clinical distribution,antimicrobial resistance,carbapenemase resistance genes,virulence genes,capsular serotypes and ST subtypes of carbapenem-resistant Klebsiella pneumoniae(CRKP)strains in intensive care unit of a tertiary hospital in Hunan Province for better management of CRKP infections.Methods CRKP strains were isolated from 8 intensive care units of the First People's Hospital of Chenzhou City from January 2020 to December 2021.The isolates were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and tested by VITEK Compact 2 for antimicrobial susceptibility.Carbapenemase phenotype was detected by modified carbapenem inactivation method(mCIM).The capsular serotypes were determined by wzi sequencing.Carbapenem resistance genes and virulence genes were identified by PCR and Sanger sequencing.The strains were also analyzed by multilocus sequence typing(MLST)in terms of ST subtypes.Results The 75 CRKP strains were mainly isolated from geriatric ICU(28.0％)and neurosurgery ICU(20.0％).Overall,6.7％(5/75)and 16.0％(12/75)of the CRKP strains were resistant to tigecycline and ceftazidime-avibactam,respectively.The CRKP strains(＞96.0％resistant)were highly resistant to carbapenems,cephalosporins,β-lactam/β-lactamase inhibitor combinations,and levofloxacin.PCR and sequencing analysis found blaKPC-2 gene in 61 strains(81.3％),blaNDM-1 gene in 11 strains(14.7％),blaNDM-5 gene in 1 strain(1.3％),and blaOXA-48 gene in 2 strains(2.7％).MLST revealed that ST11(54.7％,41/75),ST1883(13.3％,10/75),and ST307(6.7％,5/75)were the top three ST subtypes.All ST11 and ST1883 CRKP strains harbored blaKPC-2.KL64(38.7％,29/75)and KL47(25.3％,19/75)were the most prevalent capsular serotypes among the 75 CRKP strains.The most common virulence genes among these CRKP strains were rmpA2(48.0％,36/75),iroN(38.7％,23/75)and iucA(37.3％,15/75).Conclusions The CRKP strains isolated from the intensive care units were mainly ST11-KL64 and ST11-KL47 types.Most of the strains harbor blaKPC-2 and virulence gene,and associated with high level antimicrobial resistance.It is urgent to strengthen the monitoring of molecular epidemiological characteristics of CRKP in order to inform individualized and precision treatment of CRKP infections.

Tuberculosis, a global infectious disease caused by Mycobacterium tuberculosis infection, ranks among the top ten causes of human death. The BCG vaccine is currently the only tuberculosis vaccine in clinical use, but its effectiveness in protecting adults from tuberculosis remains uncertain. In recent years, protein subunit vaccines have garnered significant attention due to their well-defined composition, high safety profile, and ability to enhance the BCG vaccine′s effects. However, the protein antigens in subunit tuberculosis vaccines typically elicit weak immune responses, necessitating the use of adjuvants to boost their immunogenicity. This review focuses on exploring various types of adjuvants utilized in novel tuberculosis vaccine formulations, ranging from those undergoing clinical trials to those still in preclinical development. The review provides an objective assessment of the advantages, disadvantages, and suitability of different adjuvants, while also highlighting current challenges in adjuvant research and proposing future research directions.

Objective To observe the clinical effect of arthroscopic repair of isolated degenerative medi-al meniscus injury in patients over 45 years old and to evaluate the healing after more than two years of fol-low-up through MRI.Methods A retrospective study was conducted on the clinical effects of 51 patients with isolated degenerative medial meniscus injury who underwent arthroscopic repair from January 2016 to Decem-ber 2020.The meniscus was sutured by total internal suture or total internal suture combined with internal-to-external technique.IKDC score,Tegner score and Lysholm score were recorded before operation and at the last follow-up(at least 24 months).The type and location of meniscus injury during operation,and the microscopic classification and location of cartilage injury were recorded.At the same time,MRI was used to observe the healing of meniscus and ICRS cartilage injury grading.On MRI,0-2 degree injury was defined as meniscus healing,and 3 degree injury was defined as nonunion.The clinical repair failure was judged according to the Barrett evaluation criteria.Results The average age of the patients was(57.3±8.4)years,and the average follow-up time was(55.0±15.3)months.There were 47.1％complex fissures,31.3％horizontal fissures and 21.6％other types.The Lysholm score increased from 53.5±6.4 preoperative to 87.6±7.9 postoperative,the IKDC score increased from 33.6±4.7 preoperative to 72.8±5.3 postoperative,and the Tegner score increased from 1.0±0.1 preoperative to 4.0±0.5 postoperative.Three patients were judged as clinical repair failure,and five patients had progressed cartilage injury after surgery.At the last follow-up,the meniscus healing was observed on MRI:0 degree in two cases(3.9％),1 degree in 17 cases(33.3％),2 degree in 11 cases(21.6％),3 degree in 21 cases(41.2％).Conclusion For middle-aged and elderly patients with isolated de-generative medial meniscus injury,if conservative treatment is not effective or there are mechanical obstruc-tion factors,arthroscopic meniscus repair can achieve good clinical results and low clinical repair failure rate.