Recombinant Schistosoma japonicum egg ribonuclease SjCP1412 inhibits the activation of LX-2 hepatic stellate cells in vitro / 中国血吸虫病防治杂志

Objective To investigate the effect of recombinant Schistosoma japonicum egg ribonuclease SjCP1412 (rSjCP1412) on proliferation, cell cycle, apoptosis and activation of human hepatic stellate cells LX-2 in vitro, and explore the underlying mechanisms. Methods The rSjCP1412 protein was expressed in Escherichia coli BL21 by prokaryotic expression, and the highly purified soluble rSjCP1412 protein was prepared by Ni NTA affinity chromatography and urea gradient refolding dialysis. Yeast RNA was digested using 12.5, 25.0, 50.0 µg rSjCP1412 proteins at 37 °C for 2, 3, 4 h, and the enzymatic products were electrophoresed on 1.5% agarose gel to observe the RNAase activity of rSjCP1412 protein. The proliferation of LX-2 cells stimulated by different doses of rSjCP1412 protein for 48 hours was measured using CCK-8 assay, and the apoptosis of LX-2 cells stimulated by different doses of rSjCP1412 protein for 48 hours was detected using the Annexin V-FITC/PI double staining, while the percentage of LX-2 cells at G0/G1, S and G2/M phases of cell cycle following stimulation with different doses of rSjCP1412 protein for 48 h was detected by DAPI staining. The type I collagen, type III collagen and α-smooth muscle actin (α-SMA) mRNA expression was quantified using quantitative florescent real-time PCR (qPCR) assay and Western blotting at transcriptional and translational levels in LX-2 cells following stimulation with different doses of rSjCP1412 protein for 48 h, while soluble egg antigen (SEA) served a positive control and PBS without rSjCP1412 protein as a normal control in the above experiments. The expression of collagen I, α-SMA and Smad4 protein was determined using Western blotting in LX-2 cells following stimulation with rSjCP1412 protein, transforming growth factor-β1 (TGF-β1) alone or in combination, to examine the signaling for the effect of rSjCP1412 protein on LX-2 cells. Results The rSjCP1412 protein was successfully expressed and the highly purified soluble rSjCP1412 protein was prepared, which had a RNase activity. Compared with the normal group, the survival rates of LX-2 cells significantly decreased post-treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein and SEA for 48 h (F = 22.417 and 20.448, both P values < 0.05). The apoptotic rates of LX-2 cells significantly increased post-treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h (F = 11.350, P < 0.05), and treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h resulted in arrest of LX-2 cells in G0/G1 phase (F = 20.710, P < 0.05). Treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h caused a significant reduction in relative expression levels of collagen I (F = 11.340, P < 0.05), collagen III (F = 456.600, P < 0.05) and α-SMA mRNA (F = 23.100, P < 0.05) in LX-2 cells, and both rSjCP1412 protein and SEA treatment caused a significant reduction in collagen I (F = 1 302.000, P < 0.05), α-SMA (F = 49.750, P < 0.05) and Smad4 protein expression (F = 52.420, P < 0.05) in LX-2 cells. In addition, rSjCP1412 protein treatment inhibited collagen I (F = 66.290, P < 0.05), α-SMA (F = 31.300, P < 0.05) and Smad4 protein expression (F = 27.010, P < 0.05) in LX-2 cells activated by TGF-β1. Conclusion rSjCP1412 protein may induce apoptosis of LX-2 cells and inhibit proliferation, cell cycle and activation of LX-2 cells through down-regulating Smad4 signaling molecules.

Spinal metastases combined with leptomeningeal metastasis: A case report / 北京大学学报(医学版)

Spinal metastases (SM) is the commonest form of solid tumors osseous metastasis, for which surgical dissection is often performed when combined with spinal cord compression. Leptomeningeal metastasis (LM) results from dissemination of cancer cells to both the leptomeninges (pia and arachnoid) and cerebrospinal fluid (CSF) compartment. The spread of LM may occur via multiple routes, such as hematogenous, direct infiltration from metastatic brain lesions, or via iatrogenic seeding of CSF. Signs and symptoms associated with LM are generalized and various while early diagnosis of LM is challenging. Cytological evaluation of the CSF and gadolinium enhanced MRI brain and spine is the gold standard for diagnosing LM and CSF can help assess treatment response. While a number of other potential CSF biomarkers have been investigated both for the diagnosis as well as monitoring of LM, none have been established as a component of the standard evaluation of all LM or suspected LM patients. Management goals of LM include improving patient's neurologic function, quality of life, preventing further neurologic deterioration and prolonging survival. In many cases, it may be reasonable to pursue a palliative and comfort focused course, even from the initial LM diagnosis. Surgery is not recommended considering the risk of seeding with cerebrospinal fluid. A diagnosis of LM carries a poor prognosis with an estimated median survival of only 2-4 months despite therapy. Spinal metastases combined with leptomeningeal metastasis (SM+LM) is not uncommon and its treatment is similar to LM. LM can appear at the same time as SM or directly invaded by SM, which is thought regarding the pathophysiology of LM remains speculative and not systematically studied. The present article reports a 58-year-old woman who was first diagnosed with SM, but worsened after surgery repeated MRI examinations confirmed coexisting LM. Relevant literature was reviewed to summarize the epidemiology, clinical manifestations, imaging characteristics, diagnosis and treatment of SM+LM, so as to improve the understanding of the disease and promote early diagnosis. It should be vigilant to merge LM for the patient with SM when atypical clinical manifestations, rapid disease progression or inconsistent with imaging occurred. Repeated examinations of cerebrospinal fluid cytology and enhanced MRI should be considered when SM+LM is suspected to achieve timely adjustment of diagnosis and treatment strategy for better prognosis.

Biometric investigation of ocular components in various types of anisometropia in school-age children / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To compare the differences and correlations between different types of anisometropia, binocular visual acuity and biological parameters in school-age children.METHODS: A total of 128 school-age children(6-12 years)with mild-to-moderate anisometropia were retrospectively analyzed. Subjects were divided into five groups according to anisometropia type. All participants underwent cycloplegic refraction, A-scan ultrasound biometry, and corneal topography. Refractive status, best-corrected visual acuity(BCVA), anterior chamber depth(ACD), lens thickness(LT), vitreous chamber depth(VCD), axial length(AL), corneal radius(CR), and ratio of AL and CR(AL/CR)were recorded. Kruskal-Wallis and Spearman rank correlation tests were then used for statistical analysis.RESULTS: Hyperopic anisometropia had the greatest binocular vision difference(0.14±0.20). Myopic anisometropia had the greatest asymmetry in AL and VCD(0.56±0.41 and 0.56±0.39 mm, respectively). Anisometropia was positively correlated with BCVA, VCD, AL, and AL/CR(r=0.266, 0.379, 0.350, 0.263, respectively; P&#x0026;#x003C;0.05), and it was not significantly correlated with LT and CR(r=-0.019,-0.069, respectively; P&#x0026;#x003E;0.05), while no parameters had a statistically significant correlation with anisometropia in each group.CONCLUSION: School-age children with hyperopic anisometropia showed the greatest difference of binocular acuity in the four types of anisometropia. The inter-ocular differences of biometric parameters in simple hyperopic or myopic anisometropia were mainly attributed to the asymmetry of VCD and AL, while the differences in ocular parameters were not statistically significant in school-age children with astigmatic anisometropia.

Research advances on the role and mechanism of microRNA in hypertrophic scar / 中华烧伤杂志

Hypertrophic scar (HS) affects the function and beauty of patients, and brings a heavy psychological burden to patients. However, the specific pathogenesis mechanism of HS in molecular biology level is not yet clear, and this disease is still one of the clinical diseases difficult to prevent and cure. MicroRNA (miR) is a family of single-stranded endogenous noncoding RNAs that can regulate gene expression. The abnormal transcription of miR in hypertrophic scar fibroblasts can affect the transduction and expression of downstream signal pathway or protein, and the exploration of miR and its downstream signal pathway and protein helps deeply understand the occurrence and development mechanism of scar hyperplasia. This article summarized and analyzed how miR and multiple signal pathways involve in the formation and development of HS in recent years, and further outlined the interaction between miR and target genes in HS.

MiR-340 mediates the involvement of high mobility group box 1 in the pathogenesis of liver fibrosis / 中华肝脏病杂志

Objective: To explore the pathogenic mechanism of the miR-340/high mobility group box 1 (HMGB1) axis in the formation of liver fibrosis. Methods: A rat liver fibrosis model was established by injecting CCl(4) intraperitoneally. miRNAs targeting and validating HMGB1 were selected with gene microarrays after screening the differentially expressed miRNAs in rats with normal and hepatic fibrosis. The effect of miRNA expressional changes on HMGB1 levels was detected by qPCR. Dual luciferase gene reporter assays (LUC) was used to verify the targeting relationship between miR-340 and HMGB1. The proliferative activity of the hepatic stellate cell line HSC-T6 was detected by thiazolyl blue tetrazolium bromide (MTT) assay after co-transfection of miRNA mimics and HMGB1 overexpression vector, and the expression of extracellular matrix (ECM) proteins type I collagen and α-smooth muscle actin (SMA) was detected by western blot. Statistical analysis was performed by analysis of variance and the LSD-t test. Results: Hematoxylin-eosin and Masson staining results showed that the rat model of liver fibrosis was successfully established. Gene microarray analysis and bioinformatics prediction had detected eight miRNAs possibly targeting HMGB1, and animal model validation had detected miR-340. qPCR detection results showed that miR-340 had inhibited the expression of HMGB1, and a luciferase complementation assay suggested that miR-340 had targeted HMGB1. Functional experiments results showed that HMGB1 overexpression had enhanced cell proliferation activity and the expression of type I collagen and α-SMA, while miR-340 mimics had not only inhibited cell proliferation activity and the expression of HMGB1, type I collagen, and α-SMA, but also partially reversed the promoting effect of HMGB1 on cell proliferation and ECM synthesis. Conclusion: miR-340 targets HMGB1 to inhibit the proliferation and ECM deposition in hepatic stellate cells and plays a protective role during the process of liver fibrosis.

Mechanism of artesunate on bone destruction in experimental rheumatoid arthritis based on transcriptomics and network pharmacology / 中国中药杂志

The present study investigated the mechanism of artesunate in the treatment of bone destruction in experimental rheumatoid arthritis(RA) based on transcriptomics and network pharmacology. The transcriptome sequencing data of artesunate in the inhibition of osteoclast differentiation were analyzed to obtain differentially expressed genes(DEGs). GraphPad Prism 8 software was used to plot volcano maps and heat maps were plotted through the website of bioinformatics. GeneCards and OMIM were used to collect information on key targets of bone destruction in RA. The DEGs of artesunate in inhibiting osteoclast differentiation and key target genes of bone destruction in RA were intersected by the Venny 2.1.0 platform, and the intersection target genes were analyzed by Gene Ontology(GO)/Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment. Finally, the receptor activator of nuclear factor-κB(RANKL)-induced osteoclast differentiation model and collagen-induced arthritis(CIA) model were established. Quantitative real time polymerase chain reaction(q-PCR), immunofluorescence, and immunohistochemistry were used to verify the pharmacological effect and molecular mechanism of artesunate in the treatment of bone destruction in RA. In this study, the RANKL-induced osteoclast differentiation model in vitro was established and intervened with artesunate, and transcriptome sequencing data were analyzed to obtain 744 DEGs of artesunate in inhibiting osteoclast differentiation. A total of 1 291 major target genes of bone destruction in RA were obtained from GeneCards and OMIM. The target genes of artesunate in inhibiting osteoclast differentiation and the target genes of bone destruction in RA were intersected to obtain 61 target genes of artesunate against bone destruction in RA. The intersected target genes were analyzed by GO/KEGG enrichment. According to the results previously reported, the cytokine-cytokine receptor interaction signaling pathway was selected for experimental verification. Artesunate intervention in the RANKL-induced osteoclast differentiation model showed that artesunate inhibited CC chemokine receptor 3(CCR3), CC chemokine receptor 1(CCR1) and leukemia inhibitory factor(LIF) mRNA expression in osteoclasts in a dose-dependent manner compared with the RANKL-induced group. Meanwhile, the results of immunofluorescence and immunohistochemistry showed that artesunate could dose-dependently reduce the expression of CCR3 in osteoclasts and joint tissues of the CIA rat model in vitro. This study indicated that artesunate regulated the CCR3 in the cytokine-cytokine receptor interaction signaling pathway in the treatment of bone destruction in RA and provided a new target gene for the treatment of bone destruction in RA.

Inhibition of glutaminolysis alleviates myocardial fibrosis induced by angiotensin II / 生理学报

The present study was aimed to investigate the role and mechanism of glutaminolysis of cardiac fibroblasts (CFs) in hypertension-induced myocardial fibrosis. C57BL/6J mice were administered with a chronic infusion of angiotensin II (Ang II, 1.6 mg/kg per d) with a micro-osmotic pump to induce myocardial fibrosis. Masson staining was used to evaluate myocardial fibrosis. The mice were intraperitoneally injected with BPTES (12.5 mg/kg), a glutaminase 1 (GLS1)-specific inhibitor, to inhibit glutaminolysis simultaneously. Immunohistochemistry and Western blot were used to detect protein expression levels of GLS1, Collagen I and Collagen III in cardiac tissue. Neonatal Sprague-Dawley (SD) rat CFs were treated with 4 mmol/L glutamine (Gln) or BPTES (5 μmol/L) with or without Ang II (0.4 μmol/L) stimulation. The CFs were also treated with 2 mmol/L α-ketoglutarate (α-KG) under the stimulation of Ang II and BPTES. Wound healing test and CCK-8 were used to detect CFs migration and proliferation respectively. RT-qPCR and Western blot were used to detect mRNA and protein expression levels of GLS1, Collagen I and Collagen III. The results showed that blood pressure, heart weight and myocardial fibrosis were increased in Ang II-treated mice, and GLS1 expression in cardiac tissue was also significantly up-regulated. Gln significantly promoted the proliferation, migration, mRNA and protein expression of GLS1, Collagen I and Collagen III in the CFs with or without Ang II stimulation, whereas BPTES significantly decreased the above indices in the CFs. α-KG supplementation reversed the inhibitory effect of BPTES on the CFs under Ang II stimulation. Furthermore, in vivo intraperitoneal injection of BPTES alleviated cardiac fibrosis of Ang II-treated mice. In conclusion, glutaminolysis plays an important role in the process of cardiac fibrosis induced by Ang II. Targeted inhibition of glutaminolysis may be a new strategy for the treatment of myocardial fibrosis.

Ketogenic diet improves low temperature tolerance in mice by up-regulating PPARα in the liver and brown adipose tissue / 生理学报

The aim of the present study was to investigate the effects of short-term ketogenic diet on the low temperature tolerance of mice and the involvement of peroxisome proliferator-activated receptor α (PPARα). C57BL/6J mice were divided into two groups: normal diet (WT+ND) group and ketogenic diet (WT+KD) group. After being fed with normal or ketogenic diet at room temperature for 2 d, the mice were exposed to 4 °C low temperature for 12 h. The changes in core temperature, blood glucose, blood pressure of mice under low temperature condition were detected, and the protein expression levels of PPARα and mitochondrial uncoupling protein 1 (UCP1) were detected by Western blot. PPARα knockout mice were divided into normal diet (PPARα-/-+ND) group and ketogenic diet (PPARα-/-+KD) group. After being fed with the normal or ketogenic diet at room temperature for 2 d, the mice were exposed to 4 °C low temperature for 12 h. The above indicators were also detected. The results showed that, at room temperature, the protein expression levels of PPARα and UCP1 in liver and brown adipose tissue of WT+KD group were significantly up-regulated, compared with those of WT+ND group. Under low temperature condition, compared with WT+ND, the core temperature and blood glucose of WT+KD group were increased, while mean arterial pressure was decreased; The ketogenic diet up-regulated PPARα protein expression in brown adipose tissue, as well as UCP1 protein expression in liver and brown adipose tissue of WT+KD group. Under low temperature condition, compared to WT+ND group, PPARα-/-+ND group exhibited decreased core temperature and down-regulated PPARα and UCP1 protein expression levels in liver, skeletal muscle, white and brown adipose tissue. Compared to the PPARα-/-+ND group, the PPARα-/-+KD group exhibited decreased core temperature and did not show any difference in the protein expression of UCP1 in liver, skeletal muscle, white and brown adipose tissue. These results suggest that the ketogenic diet promotes UCP1 expression by up-regulating PPARα, thus improving low temperature tolerance of mice. Therefore, short-term ketogenic diet can be used as a potential intervention to improve the low temperature tolerance.

Influencing factors of negative conversion time of nucleic acid detection in 228 patients infected with Omicron variant of SARS-CoV-2 in Shanghai / 中国热带医学

@#Abstract: Objective To analyze and compare the effects of different clinical characteristics on the negative conversion time of nucleic acid detection after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant infection, and to provide a scientific basis for the isolation and treatment of coronavirus disease 2019 (COVID-19). Methods　The epidemiological and clinical data of 228 mild SARS-CoV-2 Omicron variant infected patients diagnosed in Shanghai were retrospectively collected from April 27, 2022 to June 8, 2022 in Wujiaochang designated Hospital, Yangpu District, Shanghai. The negative conversion time of nucleic acid detection was used as the outcome variable, and the patients were divided into A (≤18 days) and B (>18 days). Univariate and multivariate logistic regression analysis were used to analyze the influencing factors of the negative conversion time of nucleic acid detection. Results　The mean nucleic acid conversion time of 228 patients was (18.7±12.1) d, with the median time of 18 (2-46) d. Among them, 120 patients in group A had an average nucleic acid conversion time of (13.2±2.0) d, and 108 cases in group B had an average nucleic acid conversion time of (20.8±1.3) d. Univariate analysis showed that there were no statistically significant differences in the effects of hypertension, coronary heart disease, diabetes, hypokalemia, malignant tumors, neuropsychiatric diseases, chronic digestive diseases on the negative nucleic acid conversion time (P>0.05); however, there were significant differences in the effects of combined cerebrovascular disease, leukopenia, chronic respiratory system diseases and vaccination on the negative nucleic acid conversion time (P<0.05). Further multivariate logistic regression analysis revealed that the combination of chronic respiratory diseases and non-vaccination were significant risk factors for prolongation of negative nucleic acid conversion time (P<0.05). Conclusions The results of this study show that gender, age and whether hypertension, coronary heart disease, diabetes mellitus, hypokalemia, malignant tumor, neuropsychiatric disease and chronic digestive disease have no significant effect on the nucleic acid conversion time, whereas chronic respiratory disease and no vaccination are significantly correlated with the prolongation of nucleic acid conversion time in SARS-CoV-2 Omicron-infected patients.

Effects of Ranibizumab on ocular biological parameters in the treatment of retinopathy of prematurity / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the effect of intravitreal ranibizumab injection on ocular parameters in the treatment of retinopathy of prematurity(ROP), and analyze its relationship with birth weight(BW)and postmenstrual age(PMA).METHODS: A total of 98 premature infants who received routine ROP screening at Northwest Women's and Children's Hospital from January 1, 2016 to January 31, 2022 were selected, and they were divided into ROP group(49 cases)and non-ROP group(49 cases)according to the results of Retcam3 fundus screening. All children in ROP group were treated with intravitreal ranibizumab injection, with an average PMA of 38.02±3.03 weeks. The ocular parameters were measured at the PMA of 0 month(40 weeks±14d), 3 months(52 weeks±28d)and 6 months(64 weeks±28d), respectively.RESULTS: There was no difference in axial length(AL), anterior chamber depth(ACD), lens thickness(LT), vitreous length(VL)and central corneal thickness(CCT)between ROP group and non-ROP group at the PMA of 0 month(P&#x003E;0.05); At the PMA of 3 and 6 months, ACD in ROP group was higher than that in non-ROP group, and LT was lower than that in non-ROP group(P&#x003C;0.05); at the PMA of 6 months, AL and VL in ROP group were lower than those in non-ROP group(P&#x003C;0.05). AL, ACD and VL were positively correlated with PMA in ROP group and non-ROP group, while CCT was negatively correlated with PMA; there was a positive correlation between LT and PMA in children without ROP. There was no correlation among LT, BW and PMA in ROP group.CONCLUSION: The ocular development of children with early ROP(PMA 0～6 months)treated by intravitreal ranibizumab injection is slower than that of premature infants without ROP, and BW and PMA are the main influencing factors of ocular parameters of premature infants.

Determination of acetone and butanone in urine of occupational populations by headspace-gas chromatography / 预防医学

Objective @#To establish a headspace-gas chromatography ( HS-GC ) method for the determination of acetone and butanone, the biomarkers of occupational exposure to isopropanol and butanone, in urine of occupational populations. @*Methods @#Urine samples at 5.0 mL were transferred to a headspace bottle, added with 2.0 g anhydrous sodium sulfate, sealed immediately, and placed in a headspace sampler-gas chromatograph-mass spectrometer. Following heating at 60 ℃ for 30 min, 0.5 mL urine samples were injected and separated with the DB-FFAP capillary chromatographic column, and determined with the flame ionization detector. In addition, the retention time and peak area were determined. @*Results @#The peak area appeared a linear relationship with mass concentrations of acetone at 0.16-80 mg/L and butanone at 0.03-16 mg/L (correlation coefficient, 0.999 9), with detection limits of 0.009 and 0.004 mg/L, quantitation limits of 0.03 and 0.02 mg/L, respectively. The mean recovery rates of spiked samples were 93.67%-99.37% and 91.18%-94.41% for low, medium and high concentrations of acetone and butanone, and the relative standard deviations of 1.53%-3.69% and 2.54%-6.58%, respectively. @*Conclusion @#A highly sensitive and repeatable HS-GC method is successfully established for simultaneous determination of acetone and butanone in urine samples by optimizing sample pretreatment and separation, which is feasible for qualitative and quantitative analyses of acetone and butanone in urine.

Clinical and genetic spectrum of SCN2A gene associated epilepsy and episodic ataxia / 中华儿科杂志

Objective: To explore the clinical manifestations and genetic characteristics of patients with epilepsy and episodic ataxia caused by SCN2A gene variation. Methods: The clinical data of seizure manifestation, imaging examination and genetic results of 5 patients with epilepsy and (or) episodic ataxia because of SCN2A gene variation admitted to the Department of Pediatrics, the Third Affiliated Hospital of Zhengzhou University from July 2017 to January 2021 were analyzed retrospectively. Results: Among 5 patients, 4 were female and 1 was male. The onset age of epilepsy ranged from 4 days to 8 months. There were 2 cases of benign neonatal or infantile epilepsy and 3 cases of epileptic encephalopathy, in whom 1 case had development retardation,1 case transformed from West syndrome to infantile spasm and another one transformed from infantile spasm to Lennox-Gastaut syndrome. One case of benign neonatal-infantile epilepsy was characterized by neonatal onset seizures and episodic ataxia developed at the age of 78 months. Electroencephalograms at first visit of 5 cases showed that 2 cases were normal, 1 case had focal epileptic discharge, and 2 cases had multi-focal abnormal discharge with peak arrhythmia. The brain magnetic resonance imaging (MRI) of 3 cases were nomal, 1 case was abnormal (brain atrophy with decreased white matter) and the results of 1 case was unknown. The follow-up time ranged from 17 months to 89 months. Four cases of epilepsy were controlled and 1 case died at 2 years of age. Two cases had normal intelligence and motor development, 2 had moderate to severe intelligence retardation and motor critical state, and 1 had moderate to severe intelligence and motor development retardation. SCN2A gene variations were identified in all cases. There were 4 missense variations and 1 frameshift variation. Three variations had not been reported so far, including c.4906A>G,c.3643G>T,c.638delT. Conclusions: Variations in SCN2A gene can cause benign neonatal or infantile epilepsy and epileptic encephalopathy. Some children develop episodic ataxia with growing age. The variation of SCN2A gene is mainly missense variation.

Xiaoai Jiedu Prescription-containing Serum Enhances Lethal Effect of NK Cells on Colon Cancer Cells / 中国实验方剂学杂志

ObjectiveTo explore the influence of Xiaoai Jiedu prescription （XJP）-containing serum on natural killer （NK） cells′ lethal effect on colon cancer cells and the molecular mechanism. MethodXJP-containing serum （0.1%， 0.5%， 1%， 5%， 10%） was used to treat HCT-116 cells and NK-92MI cells respectively for 24 h， and methyl thiazolyl tetrazolium （MTT） assay was employed to detect cell proliferation. Then， low-concentration （0.1%， 0.5%， 1%） XJP-containing serum was selected to treat co-cultured HCT-116 cells and NK-92MI cells for 24 h and calcein acetoxymethyl ester/propidium iodide （Calcein-AM/PI） was applied to detect the killing effect of NK cells on colon cancer cells. Flow cytometry was used to detect apoptosis of colon cancer cells， Western blot the expression of apoptosis-related proteins and signal transducer and activator of transcription 4 （STAT4） pathway-related proteins， and enzyme-linked immunosorbent assay （ELISA） the secretion of interferon （IFN）-γ. ResultHigh-concentration （5%， 10%） XJP-containing serum inhibited the proliferation of HCT-116 and NK-92MI cells （P<0.01）， while low-concentration （0.1%， 0.5%， 1%） XJP-containing serum had no obvious influence on cell proliferation compared with the blank group. As compared with the blank group， low-concentration XJP-containing serum enhanced the killing activity of NK cells against colon cancer cells in a concentration-dependent manner （P<0.01）， and induced apoptosis of colon cancer cells （P<0.01）. Moreover， XJP-containing serum （0.1%， 0.5%， 1%） down-regulated the expression of B-cell lymphoma 2 （Bcl-2） and B-cell lymphoma-extra large （Bcl-xl）， and up-regulated the expression of Bcl-2-associated X （Bax） compared with the blank group （P<0.05， P<0.01）. Compared with the co-culture group， XJP-containing serum （0.1%， 0.5%， 1%） increased the expression of p-STAT4 and IFN-γ （P<0.05）. ELISA result showed that XJP-containing serum （0.1%， 0.5%， 1%） raised IFN-γ secretion （P<0.01）. ConclusionXJP-containing serum can enhance the activity of NK cells to kill colon cancer cells. The mechanism is the likelihood that it activates STAT4 pathway， increases IFN-γ secretion by NK cells， down-regulates the expression of Bcl-xl and Bcl-2， and up-regulates the expression of Bax， thereby promoting the apoptosis of colon cancer cells.

Therapeutic Effects of Mind-tranquilizing Chinese Medicines on Sleep-disorders by Regulating Energy Metabolism： A Review / 中国实验方剂学杂志

Sleep plays an important role in energy balance. As reported， sleep disorder is an important risk factor for metabolic diseases. Controlling the relationship between energy metabolism and sleep can affect sleep homeostasis and body metabolic rate. Chinese medicine， with remarkable curative effects in the prevention and treatment of insomnia， has the characteristics of green， safety， and few side effects， and attracts extensive attention of scholars in the world. In recent years， remarkable progress has been made in the research on the mechanism of Chinese medicine in interfering with sleep. This paper reviewed the research progress of mind-tranquilizing Chinese medicines， such as compounds （pterostilbene）， Chinese medicinal drugs （Ziziphi Spinosae Semen）， and Chinese medicinal prescriptions （Jiaotaiwan， Suanzaoren tang， Tianwang Buxindan， Anmeidan， Banxia Houpotang， Qihuo decoction， Songyu Anshen prescriptions， and Shuxie Yihao prescriptions） in the treatment of sleep disorders by regulating energy metabolism. The findings revealed that Chinese medicine can intervene in the sleep deprivation model by affecting metabolism-related pathways such as material metabolism， mitochondrial function， oxidative stress and inflammatory response， appetite system， and biological clock system. In terms of frequency of use， the top drugs are Ziziphi Spinosae Semen， Poria， Schisandrae Chinensis Fructus， and Salviae Miltiorrhizae Radix et Rhizoma which affect heart and liver meridians to regulate blood circulation， ensure energy supply， and play the role of nourishing the heart and tranquilizing the mind. The present paper summarized the effects and mechanisms of Chinese medicine in the treatment of insomnia and other sleep disorders from the perspective of energy metabolism to provide references for further research and exploration of diseases in the future.

Anmeidan Modulates Circadian Rhythm of Spontaneous Activity and Influences Clock Proteins in Sleep-deprived Rats / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Anmeidan （AMD） on biological rhythm and related protein expression in sleep-deprived rats. MethodA total of 80 SD rats were randomized into control group （Ctrl， equivalent volume of saline）， model group （SD， equivalent volume of saline）， AMD group （9.09 g·kg-1·d-1）， and melatonin group （MT， 0.27 g·kg-1·d-1）. Insomnia was induced in rats by self-made sleep deprivation box （4 weeks）. Circadian rhythm of spontaneous activity was evaluated by spontaneous activity video analysis system. Morphology of hypothalamus was observed based on hematoxylin-eosin （HE） staining， and the histomorphology of hypothalamus neurons and the Nissl's bodies based on Nissl staining. Western blotting was employed to detect the expression of hypothalamic proteins in cAMP-response element binding protein （CREB）/clock gene period （Per） pathway， and immunohistochemistry the expression of brain and muscle ARNT-like protein 1 （Bmal1）， Clock， Per1， and cryptochrome circadian regulator 1 （Cry1）. ResultThe model group demonstrated circadian rhythm disorder， as manifested by the significant increase in activity time in 6 designated time periods compared with the control group， and the rise in the activity speed and frequency （P<0.01）. Moreover， model group showed decrease in number of neurons which were sparsely arranged with shrunken or fragmented nuclei， reduction in number and loss of Nissl's bodies with light color， and drop in the relative expression of p-CREB and Per1， and the positive rate of Bmal1， Clock， Per1， and Cry1 （P<0.01）. Compared with model group， AMD group demonstrated reduction in time， speed， and frequency of activity （P<0.01）. Moreover， the AMD group also showed alleviation of neuronal damage （P<0.01）， and increase in the number of neurons with clear nuclei and cytoplasm in some， and the number of Nissl's bodies. AMD raised the expression of p-CREB and Per1 proteins， and the positive rate of Bmal1， Clock， Per1， and Cry1 （P<0.01）. ConclusionAMD ameliorated spontaneous circadian rhythm of sleep-deprived rats by regulating CREB/Per signaling pathway and further increasing the expression of Bmal1， Clock， Per1， and Cry1.

A Pilot Clinical Study to Investigate the Hypomethylating Properties of Freeze-dried Black Raspberries in Patients with Myelodysplastic Syndrome or Myeloproliferative Neoplasm

Myelodysplastic syndromes (MDS) and myelodysplastic/myeloproliferative neoplasms (MDS/MPN) are bone marrow disorders characterized by cytopenias and progression to acute myeloid leukemia. Hypomethylating agents (HMAs) are Food and Drug Administration-approved therapies for MDS and MDS/MPN patients. HMAs have improved patients’ survival and quality of life when compared with other therapies. Although HMAs are effective in MDS and MDS/MPN patients, they are associated with significant toxicities that place a large burden on patients. Our goal is to develop a safer and more effective HMA from natural products. We previously reported that black raspberries (BRBs) have hypomethylating effects in the colon, blood, spleen, and bone marrow of mice. In addition, BRBs exert hypomethylating effects in patients with colorectal cancer and familial adenomatous polyposis. In the current study, we conducted a pilot clinical trial to evaluate the hypomethylating effects of BRBs in patients with low-risk MDS or MDS/MPN. Peripheral blood mononuclear cells (PBMCs) were isolated before and after three months of BRB intervention. CD45 + cells were isolated from PBMCs for methylation analysis using a reduced-representation bisulfite sequencing assay. Each patient served as their own matched control, with their measurements assessed before intervention providing a baseline for post-intervention results. Clinically, our data showed that BRBs were well-tolerated with no side effects. When methylation data was combined, BRBs significantly affected methylation levels of 477 promoter regions. Pathway analysis suggests that BRB-induced intragenic hypomethylation drives leukocyte differentiation. A randomized, placebo-controlled clinical trial of BRB use in low-risk MDS or MDS/ MPN patients is warranted.

Study on the pharmacodynamic material basis of Xiangju Preparations in the treatment of rhinitis and sinusitis based on network pharmacology / 药学学报

In order to clarify the pharmacodynamic substances and mechanism of Xiangju Preparations (Xiangju Tablets, Xiangju Drops) in the treatment of rhinitis and sinusitis, the multi-level network integration analysis of "ingredients-targets-pathways" was conducted. 137 chemical constituents were identified in Xiangju Preparations by high pressure liquid chromatography-quadrupole-time of flight mass spectrometry (HPLC-QTOF/MS) for the first time. Network pharmacology analysis was performed on 59 potential active components. The results of network pharmacology analysis demonstrated that the medicinal ingredients in Xiangju Preparations included caffeic acid, senkyunolide F, rosmarinic acid, ligustilide, prim-O-glucosylcimifugin, linarin, magnolin, luteolin, senkyunolide I and gallic acid. These ingredients act on the crucial targets of tumor necrosis factor (TNF), interleukin 1B (IL1B), protein kinase B (AKT1), vascular endothelial growth factor A (VEGFA), signal transducer and activator of transcription 3 (STAT3) and participate in the regulation of advanced glycosylation end products-receptor of AGEs (AGE-RAGE), TNF, nuclear factor kappa B (NF-κB), and cyclic guanosine monophosphate-protein kinase G (cGMP-PKG) signaling pathways to effectively treat rhinitis and sinusitis. The excellent binding performance between above 10 active components and 5 key target proteins was further confirmed by molecular docking, indicating that these 10 ingredients are pharmacodynamic substances of Xiangju preparations. In conclusion, this study preliminarily clarified the effective components and mechanism of Xiangju preparations in the treatment of rhinitis and sinusitis, and provided a theoretical basis for the clinical application of Xiangju preparations.

Research progress of nano delivery system in mRNA tumor vaccines / 药学学报

Tumor vaccine is one of the most promising therapeutic strategies in tumor immunotherapy. It promotes the antigen presentation process by delivering tumor antigen and then activates the anti-tumor immune response. As a new class of vaccines, messenger RNA (mRNA) vaccines can activate the immune system to achieve the purpose of immunotherapy by delivering the mRNA sequence of a specific antigen into the body and expressing the corresponding antigen protein. Compared with traditional vaccines, mRNA vaccines have the advantages of a short production cycle, high effectiveness, and strong immunogenicity. In recent years, the application of mRNA vaccines in tumor immunotherapy has attracted widespread attention, but the instability and low delivery efficiency of mRNA limit its application. Nano delivery system can effectively solve the problem of mRNA vaccine delivery, greatly promote the research process and clinical application of mRNA tumor vaccines, and has become a hot spot in the research of mRNA vaccines. In this review, we introduced the mRNA tumor vaccines, focusing on the application of nano delivery system in mRNA tumor vaccines, in order to provide new ideas and new methods for the efficient delivery of mRNA tumor vaccines and tumor immunotherapy.

Two new labdane diterpenoids from the leaves of Callicarpa formosana Rolfe / 药学学报

Two new labdane diterpenoids were isolated from 95% ethanol extract of the leaves of Callicarpa formosana Rolfe by using silica gel column, MCI column, ODS column and HPLC. Their structures were elucidated by HR-ESI-MS, NMR and ECD spectral data. All of them are new compounds, named 13E-6β-hydroxylabda-8(17),13-dien-15-oic acid (1) and 13E-7α-hydroxylabda-8(17),13-dien-15-oic acid (2). Compounds 1 and 2 were tested for antioxidant activity, and none of them had obvious activity.

Effect analysis of trajectory screw technique in fragility fracture of pelvic ring / 中国骨伤

OBJECTIVE@#To assess the clinical efficacy of minimally invasive technology with trajectory screw fixation for fragility fractures of pelvic(FFP).@*METHODS@#A retrospective case control study was performed to analyze the clinical data of 35 patients with FFP who were treated and followed up between January 2016 and December 2019. There were 12 males and 23 females, aged from 65 to 99 years with an average of(75.4±7.8) years old. There were 13 cases of type Ⅱb, 7 cases of type Ⅱc, 8 cases of type Ⅲa, 2 cases of type Ⅲb, 2 cases of type Ⅲc, 1 case of type Ⅳb, and 2 cases of type Ⅳc according to Rommens FFP comprehensive classification. All patients received the treatment of minimally invasive technology with trajectory screws fixation. According to the different methods of anterior pelvic ring fixation, FFP patients were divided into two groups:12 cases were fixed with the pedicle screw rod system in the anterior pelvic subcutaneous internal fixator (INFIX) group;23 cases were fixed with hollow screws of the pubic symphysis, superior ramus of pubis or acetabular anterior column in the screw group. The operation time, intraoperative blood loss, intraoperative fluoroscopy times, length of hospital stay, cost of internal fixation, pre- and post-operative visual analogue scale(VAS) were compared between the two groups. The fracture reduction quality was evaluated according to the Matta criteria, and the clinical function was evaluated by the Majeed functional scoring system respectively.@*RESULTS@#All patients were followed up for 12 to 39(16.5±5.4) months after surgery. There was no statistically significant difference in the operation time, intraoperative blood loss, intraoperative fluoroscopy time, and length of hospital stay between the two groups(P>0.05). As for the cost of internal fixation, the cost of internal fixation in the screw group [2 914 (2 914, 4 371) yuan] was significantly lower than that of the INFIX group [6 205 (6 205, 6 205) yuan] (P<0.05). No significant difference was observed in the incidence of postoperative complications between the two groups (P>0.05). There was no significant difference in VAS assessment at admission, 1 week, and 3 months after surgery between the two groups(P>0.05). However, the VAS assessment at 1 week and 3 months after surgery of the two groups were significantly better than those at admission(P<0.05). There was no significant difference in the quality of fracture reduction after the operation and the efficacy evaluation at the last follow-up between the two groups(P>0.05).@*CONCLUSION@#For the treatment of fragility fractures, minimally invasive technology with trajectory screw fixation can achieve good clinical efficacy. It has the advantages of being relatively minimally invasive, less bleeding, relieving the pain. It deserves clinical application.