Magnetic separation(MS)is frequently used to enrich specific targets from biological sam-ples,which is commonly performed using antibodies coupled immunomagnetic beads(IMBs).However,due to the high cost of antibodies and difficulties in the process of releasing captured targets,IMBs have limitations for large-scale enrichment of targeted bioproducts.The specificities and affinities of aptamers towards their targets are in line with antibodies,but with characteristic of significantly lower costs of prep-aration,simpler structures,and higher chemical stability.In this study,taking advantages of the distinct adsorption features of graphene for single-stranded and double-stranded nucleic acids,we designed a novo system as bioseparation method that could display aptamers on graphene surfaces to enrich targets and then manipulate their release.This system mainly includes two units.Take CD63 aptamer as an exam-ple:double-stranded oligonucleotides scaffold attached to the graphene surface for CD63 aptamer exhibi-tion;and a single-stranded oligonucleotide complementary to the scaffold bipod sequence to desorb the captured targets from graphene.In this study,firstly,using graphene oxide(GO)paper as a graphene surface,we fluorescently labeled scaffolds or CD63 proteins and compared the fluorescence intensity difference on the GO paper before and after scaffold or CD63 protein release.Subsequently,a novo mate-rial,amino-modified graphene-shelled iron-nitrogen magnetic beads were introduced to carry the scaffold,and used to capture CD63 proteins from cell lysates.The results indicate that this scaffold can display the aptamer on the graphene surface for CD63 protein capture and controlled release.Using selected modified graphene magnetic beads carried with the scaffold,we achieved the capture of CD63 proteins from cell ly-sates.This system is expected to enrich various proteins such as CD63,or capture exosomes by hooking membrane proteins.

Objective:To investigate the effect of early postoperative enteral nutrition(EN)on postoperative rehabilitation and inflammation after laparoscopic radical gastrectomy for gastric cancer,in order to provide reference for postoperative rehabilitation of such patients.Methods:Patients who received laparoscopic assisted radical gastrectomy in Department of Gastrointestinal Surgery of The First Affiliated Hospital of Bengbu Medical College from January 2020 to December 2022 were included in the analysis.According to the different ways of postoperative nutritional treatment,patients were divided into the observation group(early postoperative EN group)and the control group(parenteral nutrition group),and indexes such as postoperative rehabilitation,abdominal drainage flow and the level of inflammatory mediators in drainage fluid were compared between the two groups.Results:A total of 81 patients were included,including 41 in the observation group and 40 in the control group.Interval of the first postoperative exhaust(t=3.806;P＜0.001)and resuming diet day(t=5.510;P＜0.001),and length of postoperative hospital stay(t=2.401;P=0.019)in the observation group were shorter than those in the control group.Levels of peripheral blood albumin(t=14.040;P＜0.001)and prealbumin(t=9.832;P＜0.001)of the observation group at postoperative day(POD)5 were significantly higher than those of the control group,but there was no significant difference in hemoglobin level(t=1.477;P=0.144).The level of CRP in peripheral blood of the observation group at POD 5(t=7.758;P＜0.001)and the incidence of postoperative SIRS[(12.2％,5/41)vs(32.5％,13/40),x2=4.830;P=0.028)]were significantly lower than those in the control group.The average drainage volume(t=6.858;P＜0.001),drainage removal time(t=5.016;P＜0.001),and TNF-α level(t=4.993;P＜0.001)and IL-6 level(t=20.640;P＜0.001)in postoperative drainage at POD 5 were significantly lower in the observation group than those in the control group.Conclusion:Early postoperative EN could accelerate the rehabilitation process after laparoscopic radical gastrectomy,improve postoperative nutritional status,and reduce abdominal inflammation.

Objective:To investigate the mechanism of Xiangsha Liujunzi Tang in improving liver lipid deposition in ApoE^-/- atherosclerotic （AS） mice by affecting long noncoding RNA-HC （Lnc-HC）/microRNA-130b （miR-130b） in the regulation of cholesterol metabolism. Method:Totolly 10 C57BL/6J mice were selected as normal controls， and 30 healthy ApoE^-/- mice fed with high fat diet for 12 weeks were then randomly divided into the model group， Xiangsha Liujunzi Tang group（19.12 g·kg^-1·d^-1） and simvastatin group（2.275 mg·kg^-1·d^-1）， with gavage administration for 4 weeks. The serum lipid level of mice was detected by automatic biochemistry analyzer， and the histopathological changes of liver cells were observed by hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect expression of long noncoding RNA-HC， and miR-130b. Real-time PCR and Western blot assay were used to detect gene and protein expression of peroxisome proliferator-activated receptor gamma （PPARγ）， liver X receptor （LXR）， ATP-binding cassette transporters A1 （ABCA1）， ATP-binding cassette transporters G1 （ABCG1）， ATP-binding cassette transporters G5 （ABCG5）， and ATP-binding cassette transporters G8 （ABCG8）. Result:Compared with the normal control group， the mice in the model group showed abnormal blood lipids， larger liver cells， obvious fat vacuoles， significantly increased expression of Lnc-HC， miR-130b in liver， and significantly decreased gene and protein expression of PPARγ， LXR， ABCA1， ABCG1， ABCG5， and ABCG8 in mice liver （P<0.05，P<0.01）. Compared with the model group， the abnormal blood lipid levels of the mice in the Xiangsha Liujunzi Tang group and the simvastatin group were improved， and the number of fatty vacuoles of liver cells was significantly reduced， the expression of liver Lnc-HC， miR-130b in Xiangsha Liujunzi Tang group decreased significantly （P<0.05，P<0.01）， the gene and protein levels of liver PPARγ， ABCA1， ABCG1， ABCG5， ABCG8 in mice of the Xiangsha Liujunzi Tang group and the simvastatin group showed an upward trend. Among them， the gene and protein expression of LXR protein in the liver of the Xiangsha Liujunzi Tang group was significantly up-regulated （P<0.05）. Conclusion:Xiangsha Liujunzi Tang may improve the lipid deposition in the liver of ApoE^-/- AS mice by affecting Lnc-HC/miR-130b to regulate the cholesterol metabolism process mediated by PPARγ， thus playing a role in preventing and treating AS.

OBJECTIVE: To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province. METHODS: The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017. RESULTS: The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01). CONCLUSIONS Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.
Anti-Bacterial Agents ; Birth Weight ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Intensive Care Units, Neonatal ; Surveys and Questionnaires

Objective: To study the effects of Fengshi Qutong capsule (FSQTC) on proliferation, migration, adhesion, invasion and secretion of human synovial cells in rheumatoid arthritis (RA) induced by tumor necrosis factor-α (TNF-α) and explore its mechanism. Method: Human synovial cells (MH7A) in RA patients were induced in vitro by using TNF-α (20 μg·L^-1). After treatment with different concentrations of FSQTC (0.02,0.1,0.5 μg·L^-1), MTT colorimetric assay, transwell migration, adhesion and invasion tests were used to detect the proliferation, migration, adhesion and invasion of the MH7A, respectively. The expression levels of vascular endothelial growth factor (VEGF) and interleukin-1β (IL-1β) in MH7A supernatant were detected by enzyme linked immunosorbent assay (ELISA). Result: As compared with blank control group, TNF-α (20 μg·L^-1) significantly increased the proliferation, migration, adhesion, invasion and secretion of IL-1β and VEGF of MH7A cells (P<0.01). FSQTC (0.02,0.1,0.5 μg·L^-1) had no significant effect on proliferation of TNF-α-induced MH7A cells after treatment for 24 hours. After 48 hours of treatment, proliferation of MH7A cells induced by TNF-α was decreased in a concentration-dependent manner (P<0.05,P<0.01). Within 24 hours, the migration, adhesion, invasion, invasion, and secretion of IL-1 and VEGF in MH7A cells were also decreased significantly (P<0.05,P<0.01). Conclusion: FSQTC can inhibit the proliferation, migration, adhesion, invasion and secretion of IL-1β and VEGF in MH7A cells.

Objective:To observe the effect of Fengshi Qutong capsule (FSQTC) on protein kinase B(Akt) and mitogen-activated protein kinase (MAPK) signaling pathways of rheumatoid arthritis (RA). Method:Collagen-induced arthritis (CIA) was induced in SD rats, and the synovial membranes of the knee joints were prepared after 19 days of oral administration of 0.25, 0.5, 1 g·kg^-1 FSQTC. MH7A cells were induced by tumor necrosis factor-α (TNF-α, 20 μg·L^-1) in vitro, and human umbilical vein endothelial cells (HUVEC) were induced by vascular endothelial growth factor (VEGF). FSQTC (0.02, 0.1, 0.5 μg·L^-1) were added to MH7A/HUVEC cells, and then the cells were collected. Proteins of synovial tissue, MH7A and HUVEC cells were extracted, and then were detected the expresstion of p-Akt, p-p38 MAPK, p-extracellular signal-regulated kinase(ERK) and p-Jun n-terminal kinase(JNK) by Western blot. Result:The expression levels of p-Akt, p-p38 MAPK, p-ERK and p-JNK in the synovial membrane of CIA model were significantly increased compared with normal group (P<0.01), and the treatmend of 0.25, 0.5 and 1 g·kg^-1·d^-1 FSQTC significantly decreased their expression levels (P<0.05, P<0.01). To compared with control group, the expression levels of p-Akt, p-p38 MAPK, p-ERK and p-JNK in MH7A or HUVEC cells induced by TNF-α or VEGF were increased (P<0.01), respectively, and these factors are significantly reduced by 0.02, 0.1, 0.5 μg·L^-1 FSQTC (P<0.05, P<0.01). Conclusion:FSQTC can down-regulate the abnormal activation of Akt and MAPK signaling pathways in the synovial membrane of CIA rats, fibroblast synovial cells and vascular endothelial cells, which is related to the inhibition of synovial angiogenesis in the treatment of RA.

Objective: To study the effect of Fengshi Qutong capsule on the migration, adhesion, invasion and tube formation of human synovial cells and the phosphorylation and protein expression of vascular endothelial growth factor receptor 2 (VEGFR2). Method: With human umbilical vein endothelial cells (HUVEC) as the research object, low, middle and high-dose Fengshi Qutong capsule(0.02,0.1,0.5 μg·L^-1) on HUVEC was determined by methye thiazolye telrazlium (MTT) colorimetric assay for the follow-up experiment. The transwell migration, adhesion and transwell invasion test were used to detect the migration and adhesion of the different concentrations of Fengshi Qutong capsule in HUVEC. The expression of VEGFR2 in HUVEC was detected by Western blot, and Real-time PCR was used to detect the content of VEGFR2 mRNA in cells. Result: Compared with normal group, the proliferation of HUVEC was significantly increased after 24 h and 48 h of VEGF induction (P<0.01), the migration, adhesion, invasion and lumen formation of HUVEC were significantly increased within 24 h (P<0.01), and VEGFR2 phosphorylation protein and mRNA expression levels in HUVEC were significantly elevated in HUVEC (P<0.01). Compared with VEGF group, 0.02, 0.1 and 0.5 μg·L^-1 Fengshi Qutong capsule were administered in vitro for 48 h to inhibit HUVEC proliferation activity in a dose-dependent manner (P<0.05, P<0.01). Within 24 h, VEGF-induced HUVEC migration, adhesion, invasion and lumen formation were significantly inhibited (P<0.05), and negatively regulated VEGFR2 phosphorylation protein and gene expression levels (P<0.05). Conclusion: Fengshi Qutong capsule can inhibit the migration, adhesion, invasion and tube formation of HUVEC. This effect may be related to the inhibition of phosphorylation, and protein and mRNA expression level of VEGFR2.

Objective:To observe the intervention effect of Fengshi Qutong capsule on collagen-induced arthritis (CIA) in rats. Method:SD rats were randomly divided into normal group, model group, low, medium and high-dose Fengshi Qutong capsule groups (0.25, 0.5, 1 g·kg^-1·d^-1), and methotrexate group(0.2 mg·kg^-1·d^-1).Except for normal group, the other groups were immunized with type Ⅱ collagen and incomplete Freund's adjuvant to establish a CIA model. On the 1^st day after the first immunization, the administration group was given intragastric administration, once a day, for 19 days; on the 8^th day after the first immunization, the symptoms of joint swelling and malformation of the rats were observed, and the clinical scores and incidence of arthritis were evaluated. On the 19^th day, micro-computed tomography and bone metrology were performed, and histopathological examination of inflammatory joints was performed,andsynovial inflammation,vasospasm, cartilage erosion and bone destruction by pathological severity scores, serum interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF),matrix metalloproteinase-3 (MMP-3) and receptor activator of nuclear factor kappa B ligand(RANKL)were detected by enzyme linked immunosorbent assay. Result:Fengshi Qutong capsule could improve the symptoms of inflammatory joint redness, swelling and deformity in CIA rats in a dose-dependent manner. Compared with normal group, clinical score and incidence, joint synovial inflammation, vasospasm, cartilage erosion and pathological score of bone destruction in joint group were significantly increased (P<0.01), bone mineral density (BMD), bone volume/total volume (BV/TV), trabecular thickness (Tb. Th), trabecularnumber (Tb. N) were significantly decreased (P<0.01), bone surface area/bone volume (BS/BV) and trabecular separation (Tb. Sp) were significantly elevated (P<0.01),and levels of IL-1β, TNF-α, VEGF, MMP-3 and RANKL in serum were increased (P<0.01); compared with model group, clinical scores and incidence, joint synovial inflammation, vasospasm, cartilage erosion and bone destruction of each drug groupwere significantly lower (P<0.05, P<0.01), BMD, BV/TV, Tb.Th, Tb.N were significantly elevated (P<0.05, P<0.01), BS/BV and Tb.Sp were significantly decreased (P<0.05, P<0.01), and levels of serum IL-1β, TNF-α, VEGF, MMP-3 and RANKL were significantly decreased (P<0.05, P<0.01). And the control drug methotrexate had similar effects to the high-dose group. Conclusion:Fengshi Qutong capsule can effectively alleviate the clinical symptoms and conditions of experimental rheumatoid arthritis in rats, reduce the incidence, and relieve the histopathology and imaging severity, while inhibiting the inflammatory cytokines.

Objective: To observe the changes of dysfunctional high density lipoprotein cholesterol (dyHDL) and the intervention effect of Xiangsha Liu Junzitang in rats with spleen deficiency and hyperlipidemia, and reveal the effect and mechanism of Xiangsha Liu Junzitang on dyHDL in rats with spleen-deficiency hyperlipidemia. Method: Seventy-five SPF SD rats were randomly divided into normal group, high fat group, spleen deficiency and high fat group, Xiangsha Liu Junzitang low and high dose groups (5.67, 11.34 g·kg^-1). In the spleen deficiency and high fat group, as well as Xiangsha Liu Junzitang low and high dose groups, composite method of improper diet and exhaustive swimming was used for 15 days for modeling. After modeling for 15 days, normal group was fed with basic diet, while the high-fat group, spleen-deficiency and high-fat group, the Xiangsha Liu Junzitang low and high dose groups were fed with high-fat diet. After 12 weeks, the Xiangsha Liu Junzitang low dose and high dose groups received corresponding dosage of drugs, while normal group, high fat group and spleen deficiency high fat group received corresponding volume of normal saline. After 4 weeks, the contents of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol cholesterol (LDL-C), and high density lipoprotein cholesterol(HDL-C)were detected by automatic biochemistry analyzer, while D-xylose excretion rate was measured by phloroglucinol method. The morphological changes of liver cells were observed by hematoxylin eosin (HE) staining. The level of PON1, apoA1 and SAA in plasma were detected by enzyme-linked immunosorbent assay (ELISA). Paraoxonase 1(PON1), apolipoprotein A1 (apoA1) and serum amyloid protein A (SAA) gene expression in rats liver were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Result: As compared with normal group, the serum TC, TG, and LDL-C levels were significantly increased in the high-fat group and spleen-deficiency high-fat group (P<0.05), and HDL-C levels were significantly lower (P<0.05); the levels of PON1 and apoA1 in plasma were decreased (P<0.05), while the content of SAA was increased (P<0.05); the expression of SAA gene in liver tissues was increased (P<0.01), and the expression levels of PON1 and apoA1 genes in liver tissues were significantly decreased (P<0.01). In high-fat group and the spleen-deficiency and high-fat group, the hepatocytes were rounded and scattered, and scattered fat vacuoles were observed. In addition, the urinary D-xylose excretion rate was significantly decreased in the spleen-deficiency and high-fat group (P<0.05). After the intervention with Xiangsha Liu Junzitang, the serum TC, LDL-C levels were significantly decreased (P<0.05); HDL-C levels were significantly increased (P<0.05); plasma PON1, apoA1 levels were increased (P<0.05); the content of SAA was decreased (P<0.05); the expression of SAA gene in liver tissues was decreased (P<0.05); the expression of apoA1 gene was increased significantly (P<0.05). Liver cells swelling was significantly alleviated and fat foaming was reduced. As compared with high-fat group, the plasma PON1 and SAA levels in the spleen-deficiency high-fat group were significantly lower (P<0.05); hepatocyte swelling was obvious and foaming was aggravated. Conclusion: The lipid disorder in hyperlipidemia rats was aggravated by the spleen deficiency, but was corrected after intervention with Xiangsha Liu Junzitang. and its mechanism may be related to the regulation of the expression of dyHDL-related genes and protein.

To observe the effect of Fengshi Qutong Capsules(FSQTC) on angiogenesis of rat aortarings and in knee joint synovium of type Ⅱ collagen-induced arthritis(CIA) rats. The blood vessel of aorta rings of normal SD rats were induced by vascular endothelial growth factor(VEGF) 20 μg·L~(-1 )in vitro, and were treated with FSQTC(0.02, 0.1 and 0.5 μg·L~(-1)) continuously for 9 days. The number, length and area of neovascularization of the vascular ring were measured. SD rats were immunized to establish collagen-induced arthritis. CIA rats were treated with FSQTC(0.25, 0.5, 1 g·kg~(-1)·d~(-1)) and methotrexate(0.2 mg·kg~(-1)·d~(-1)) daily for 19 days. Histopathological examination(HE) was performed to observe the vascular morphology and vascular density in the synovial membrane of the inflamed joint. Immunohistochemistry was performed to observe the expression of platelets-endothelial cell adhesion molecule(CD31), VEGF and VEGF receptor 2(VEGFR_2)in the synovium. Immunofluorescence was performed to observe the expression of CD31 and α smooth muscle actin(αSMA) in synovial membrane.TGF-β, PDGF and VEGFR_2 in serum were detected by enzyme-linked immunosorbent assay. The number, branch length and area of blood vessels of aorta rings were significantly increased induced by VEGF, and FSQTC could significantly reduce the number, branch length and area of blood vessels. Compared with the normal group, the vascular density, CD31 positive expression, CD31~+/αSMA~- immature and total vascular positive expression in the synovial membrane of the model group were significantly increased, and so as VEGF and VEGFR_2 in the synovium. The VEGFR_2, TGF-β and PDGF in sera were also significantly increased in model group. FSQTC reduced the synovial vascular density and inhibited the positive expression of CD31, CD31~+/αSMA~- immature blood vessels and total vascular. FSQTC has no significant effect on CD31~+/αSMA~+mature blood vessels. FSQTC also negatively inhibited the expression of VEGF, VEGFR_2, TGF-β and PDGF in synovial membrane and/or sera. The effect of methotrexate is similar with to the high dose group. Our results demonstrated that FSQTC could inhibit the angiogenesis of synovial tissue in CIA rats and of aortaring in rats, which is related to the reduction of angiogenesis regulatory factor.
Animals ; Aorta ; Arthritis, Experimental ; chemically induced ; drug therapy ; Capsules ; Collagen Type II ; Drugs, Chinese Herbal ; pharmacology ; Neovascularization, Pathologic ; drug therapy ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane ; blood supply ; Vascular Endothelial Growth Factor A