Puerarin reduces impairment of intestinal and adipose immune responses to influenza virus infection in mice.

Influenza is an acute viral respiratory disease that can also cause gastroenteritis-like symptoms, such as abdominal pain, nausea, vomiting, and diarrhea. Immune dysfunction of adipose tissue is involved in the occurrence and prognosis of influenza viral pneumonia. In this study, we analyzed intestinal and adipose immune responses in mice infected with influenza virus and found that the impairment of intestinal and adipose immunity to influenza virus infection could be reversed by treatment with puerarin, a medicinal compound isolated from Pueraria lobata (called "gegen" in Chinese). We found that the lungs, small intestines (duodenum, ileum, jejunum) and large intestines (colon and rectum) of infected mice showed obvious inflammatory lesions, with significantly increased levels of virus, inflammatory cytokines (interleukin [IL]-6, IL-17, and tumor necrosis factor-α), Toll-like receptors 3, 4, and 9, and integrin αvß3 and α4, and a decreased level of secreted IgA compared to the normal control group (NC) (P < 0.05-0.001). Influenza virus infected mesenteric lymph nodes and adipose tissue, and adipokines (leptin, visfatin, "chemerin", and adiponectin) of lung and mesenteric adipose tissue were dysregulated. Puerarin treatment reversed the impairment of the intestinal and adipose immune responses in mice infected with influenza virus. Our findings suggest that influenza virus can infect adipose tissue and lead to intestinal adipose immune dysfunction in normal-weight mice and that the impairment of the intestinal and adipose immune response to influenza virus infection can be reversed by puerarin treatment.

A potential antiviral activity of Esculentoside A against binding interactions of SARS-COV-2 spike protein and angiotensin converting enzyme 2 (ACE2).

The recent emergence of the novel coronavirus (SARS-CoV-2) has resulted in a devastating pandemic with global concern. However, to date, there are no regimens to prevent and treat SARS-CoV-2 virus. There is an urgent need to identify novel leads with anti-viral properties that impede viral pathogenesis in the host system. Esculentoside A (EsA), a saponin isolated from the root of Phytolacca esculenta, is known to exhibit diverse pharmacological properties, especially anti-inflammatory activity. To our knowledge, SARS-CoV-2 uses angiotensin converting enzyme 2 (ACE2) to enter host cells. This is mediated through the proteins of SARS-CoV-2, especially the spike glycoprotein receptor binding domain. Thus, our primary goal is to prevent virus replication and binding to the host, which allows us to explore the efficiency of EsA on key surface drug target proteins using the computational biology paradigm approach. Here, the anti-coronavirus activity of EsA in vitro and its potential mode of inhibitory action on the S-protein of SARS-CoV-2 were investigated. We found that EsA inhibited the HCoV-OC43 coronavirus during the attachment and penetration stage. Molecular docking results showed that EsA had a strong binding affinity with the spike glycoprotein from SARS-CoV-2. The results of the molecular dynamics simulation revealed that EsA had higher stable binding with the spike protein. These results demonstrated that Esculentoside A can act as a spike protein blocker to inhibit SARS-CoV-2. Considering the poor bioavailability and low toxicity of EsA, it is suitable as novel lead for the inhibitor against binding interactions of SARS-CoV-2 of S-protein and ACE2.

Antiviral activity of puerarin as potent inhibitor of influenza virus neuraminidase.

Puerarin is a major isofiavone compound isolated from the root of Pueraria lobata. It was reported that puerarin had antioxidant, antiinflammatory, antitumor, cholesterol lowering, liver protective, and neuroprotective properties. However, few studies have explored the antiviral effect of puerarin and its target mechanism related to influenza virus. Here, the antiinfluenza activity of puerarin in vitro and in vivo and its mode of action on the potential inhibition of neuraminidase (NA) were investigated. Puerarin displayed an inhibitory effect on A/FM/1/1947(H1N1) (EC50 = 52.06 µM). An indirect immunofluorescence assay indicated that puerarin blocked the nuclear export of viral NP. The inhibition of NA activity confirmed that puerarin can block the release of newly formed virus particles from infected cells. Puerarin (100 and 200 mg/kg/d) exhibited effective antiviral activity in mice, conferring 50% and 70% protection from death against H1N1, reducing virus titers, and effectively alleviating inflammation in the lungs. The molecular docking results showed that puerarin had a strong binding affinity with NA from H1N1. The results of the molecular dynamics simulation revealed that puerarin had higher stable binding at the 150-loop region of the NA protein. These results demonstrated that puerarin acts as a NA blocker to inhibit influenza A virus both in cellular and animal models. Thus, puerarin has potential utility for the treatment of the influenza virus infection.

Comparative analysis of different feeder layers with 3T3 fibroblasts for culturing rabbits limbal stem cells.

AIM: To explore the possibility of human umbilical cord mesenchymal stem cells (hUCMSCs), human umbilical vein endothelial cells (hUVECs), human dental pulp stem cells (hDPSCs) and human periodontal ligament stem cells (hPDLSCs) serving as feeder cells in co-culture systems for the cultivation of limbal stem cells. METHODS: Different feeder layers were cultured in Dulbecco's modified Eagle's medium (DMEM)/F12 and were treated with mitomycin C. Rabbits limbal stem cells (LSCs) were co-cultured on hUCMSCs, hUVECs, hDPSCs, hPDLSCs and NIH-3T3, and then comparative analysis were made between each group to see their respective colony-forming efficiency (CFE) assay and immunofluorescence (IPO13,CK3/12). RESULTS: The efficiency of the four type cells in supporting the LSCs morphology and its cellular differentiation was similar to that of NIH-3T3 fibroblasts as demonstrated by the immunostaining properties analysis, with each group exhibiting a similar strong expression pattern of IPO13, but lacking CK3 and CK12 expression in terms of immunostaining. But hUCMSCs, hDPSCs and hPDLSCs feeder layers were superior in promoting colony formation potential of cells when compared to hUVECs and feeder-cell-free culture. CONCLUSION: hUCMSCs, hDPSCs and hPDLSCs can be a suitable alternative to conventional mouse NIH-3T3 feeder cells, so that risk of zoonotic infection can be diminished.

Three-Dimensional Carbon Nanotubes Forest/Carbon Cloth as an Efficient Electrode for Lithium-Polysulfide Batteries.

The development of a three-dimensionally flexible, large-surface area, high-conductivity electrode is important to improve the low conductivity and utilization of active materials and restrict the shuttle of long-chain polysulfides in Li-polysulfide batteries. Herein, we constructed an integrated three-dimensional carbon nanotube forest/carbon cloth electrode with heteroatom doping and high electrical conductivity. The as-constructed electrode provides strong trapping on the polysulfide species and fast charge transfer. Therefore, the Li-polysulfide batteries with as-constructed electrodes achieved high specific capacities of ∼1200 and ∼800 mA h g-1 at 0.1 and 1 C, respectively. After 300 cycles at 0.5 C, a specific capacity of 623 mA h g-1 was retained.

[Relationship between electron transfer capacity and fluorescence characteristics of dissolved organic matter].

This study demonstrated the relationship between redox activity and fluorescence characteristics of different dissolved organic matter (DOM) employing a combined method of electrochemistry and fluorescence spectroscopy. Chronoamperometry was used to quantitatively evaluate the electron transfer capacity (ETC) of DOM. Data showed that the electron accepting capacity of DOM ranged from 635.6 micromol x (g x C)(-1) to 1049.3 micromol x (g x C)(-1), and the electron donating capacity ranged from 27.3 micromol x (g x C)(-1) to 42.3 micromol x (g x C)(-1). For all DOM tested, the cyclic voltammetries (CVs) showed an evident pair of redox peak with the redox potentials in the range from -731 mV to -996 mV (vs. Ag/AgCl), suggesting their electrochemical activity. Using the multi-potential steps method, the ETC was measured to be 232.1-897.2 micromol x (g x C)(-1) and the electron recycling rates (ERRs) were 36.7%-78.2%, indicating the electron transfer of DOM has a reversible character. Using fluorescence excitation-emission spectroscopy (EEMs), the ERRs were found to be highly correlated with the fulvic-like fluorescence intensity of DOM with a correlation coefficient of 0. 92. This study can potentially provide a scientific base for understanding the roles of DOM in the elements cycles, pollutants degradation and biogeochemical cycles.

[Cd uptake in rice cultivars and Cd fractions in soil treated with organic acids and EDTA].

A pot experiment was conducted to examine the yield, quality and cadmium (Cd) uptake in different rice cultivars, and Cd speciation in soil after exposing to Cd (0, 1 and 5 mg x kg(-1)) in the presence of organic acids and ethylenediamine tetraacetic acid (EDTA). The results showed that general increase in the yield for cultivars Xiushui63 and II you527 was observed. Yield of two rice cultivars were in order of organic acids treatment or organic acids + 1/2EDTA treatment > EDTA treatment. The exchangeable, carbonate related and ferric-manganese oxidation related Cd increased; while organic complexation Cd and residules decreased in the presence of organic acids and EDTA. Cadmium concentrations in grain, straw and roots of both cultivars markedly reduced in the presence of organic acids and EDTA. Grain Cd concentration was the lowest for plants treated with EDTA, followed by organic acids + 1/2EDTA, and the highest Cd concentration in grain was found in the treatment with organic acids. Grain Cd concentration decreased by 9.0% to 49.3% and 16.5% to 30.6% at 1 mg x kg(-1) Cd in the presence of organic acids and EDTA, and by 12.7% to 28.5% and 4.3% to 19.1% at 5 mg x kg(-1) Cd. Cadmium concentration and accumulation in plants and total Cd content in soil were higher in Xiushui63 than in that in II you527. Grain Cd concentration decreased, and yield and quality of two rice cultivars increased at the same time in the presence of organic acids + 1/2EDTA.

[Effect of phosphor on accumulation and chemical forms of cadmium, and physiological characterization in different varieties of Capsicum annuum L].

Pot experiments were carried out to investigate the influence of different Phosphor (P) levels (0, 0.3% and 0.5%) on the plant growth, activities of antioxidant enzymes, accumulation and chemical forms of cadmium (Cd) in Capsicum annuum L. when exposed to Cd (10 mg x kg(-1)). The results showed that dry weights of leaf, fruit, roots and total dry weights of plant, and concentration and accumulation of Cd significantly differed between two varieties of Capsicum annuum L. Dry weights of fruit and total plant of Chaotianjiao increased by P (0.3% and 0.5%), while that of Yanjiao425 was inhibited. Activities of catalase (CAT) were increased at first, and then reduced in the presence of P; Activities of superoxide dismutase (SOD) and peroxidase (POD) of Chaotianjiao increased with increasing levels of P, but activities of SOD and POD of Yanjiao425 decreased with increasing levels of P. Chemical forms of Cd in fruit of Capsicum annuum L. were in order of F(NaCl) > F(HAC) > F(E) > Fr > F(HC) > F(W). The total extractable Cd, ethanol-extractable Cd, hydrochloric acid-extractable Cd and residual Cd in fruit of Ynajiao425 obviously decreased in the presence of P compared to the control, while the total extractable Cd, water-extractable Cd, acetic acid-extractable Cd and residual Cd in fruit of Chaotianjiao increased. Cadmium accumulations of Capsicum annuum L. were in order of roots > stew > leaf > fruit. Cadmium accumulations in fruit and plant of Yanjiao425 were decreased by 47.7% and 58.5% , 5.5% and 13. 1% in the presence of 0.3% and 0.5% P when exposed to Cd, and Cd accumulations in fruit and plant of Chaotianjiao were decreased by 23.6% in the presence of 0.3% P.

[The clinical characteristics and trend of conversion to type 2 diabetes mellitus of individuals with normal glucose tolerance-hyperinsulinemia].

OBJECTIVE: To study the outcomes and influencing factors of the conversion from normal glucose tolerance-hyperinsulinemia (NGT-HINS) to diabetes in the population of a community in Beijing. METHODS: All the subjects investigated received 75 g oral glucose tolerance test (OGTT) for diabetes screening carried out in May, 2006 and May, 2008. Data were calculated to analyze the outcomes and influencing factors of the conversion. HINS was diagnosed if fasting serum insulin≥15 mIU/L and/or 2-hour serum insulin after glucose loading≥80 mIU/L. RESULTS: The prevalence of NGT-HINS in the community in 2006 and 2008 was 5.28% and 8.67% (P<0.01) respectively and that of diabetes mellitus (DM) and impaired glucose regulation (IGR) was 3.52%, 6.56% in 2006 and 4.42%, 6.47% in 2008. The probability of the conversion from NGT-HINS to IGR and DM was 18.6% and 2.3%, being much higher than that from normal glucose tolerance-normoinsulinemia (NGT-NINS) (5.4% and 0.7%, P<0.01). However, the probability of the conversion from NGT-HINS to DM was 2.3%, which was much lower than that from IGR (26.3%, P<0.01). The reason might be that individuals with NGT-HINS had a higher waist circumference, BMI, fasting plasma glucose, 2 h plasma glucose and TG but a lower HDL-C than individuals with NGT-NINS in 2006. The HOMA ß-cell function index/HOMA insulin resistance index (HBCI/IR) of individuals with NGT-HINS was much lower than that of individuals with NGT-NINS, but much higher than that of individuals with IGR. Logistic regression analysis showed that age, TG and HBCI/IR were the major influencing factors of the conversion from NGT to glucose metabolic disorders. CONCLUSIONS: The probability of conversion from NGT to DM was increased remarkably when HINS was diagnosed. The reason might be that individuals with NGT-HINS suffered more metabolic risk factors and had a decreased ß-cell function. Therefore, individuals with NGT-HINS should be paid attention to in diabetes prevention study.

[Effect of different zinc levels on accumulation and chemical forms of cadmium, and physiological characterization in Capsicum annuum L].

Pot experiments were carried out to investigate the influence of different zinc (Zn) levels (0, 100, 200, 400 and 600 micromol x L(-1)) on the plant growth,activities of antioxidant enzymes, contents of chlorophyll a and b, accumulation and chemical forms of cadmium (Cd) in Capsicum annuum L. when exposed to Cd (20 mg x kg(-1)). The results showed that dry weights of leaf, stem, fruit and root, and contents of chlorophyll a and b in Capsicum annuum L. were increased by Zn ( < or = 400 micromol x L(-1)), while inhibited by high Zn (600 micromol x L(-1)). Activities of superoxide dismutase (SOD) and catalase (CAT) were reduced by Zn ( < or =400 micromol x L(-1)), the lowest activities of SOD and CAT were recorded in 400 micromol x L(-1) Zn, but activities of SOD and CAT were increased when Zn >400 micromol x L(-1). Cadmium concentrations in stem, fruit and root of Capsicum annuum L. were decreased by 2.7%-5.4%, 7.5%-28.1% and 7.6%-21.8% in the presence of Zn when exposed to Cd. The total extractable Cd, NaCl- extractable Cd, water-extractable Cd and ethanol-extractable Cd in fruit were reduced by 7.7%-21.8%, 4.11%-23.6%, 54.5%-66.8% and 4.8%-86.7% in the presence of Zn,while acetic acid- extractable Cd and residual Cd were increased by 28.0%-68.0% and 12.6%-25.0%.

[Expression of Hsp70 Grp94 and IgG in human lung carcinoma].

AIM: To explore the expression and significance of heat shock protein 70 (Hsp70), glucose regulated protein 94 (Grp94) and immunoglobulin G (IgG) in human lung carcinoma. METHODS: The expression of Hsp70, Grp94 and IgG in 40 human lung carcinomas was studied using immunohistochemical technique and image analysis. The localization among Hsp70, Grp94 and IgG was analyzed by double labeling immunofluorescent staining and laser scanning confocal microscopy. RESULTS: Hsp70, Grp94 and IgG in Human lung carcinomas showed high expression. The positive rate of Hsp70, Grp94 and IgG was 65% (26/40), 45% (18/40), and 82.5% (33/40), respectively. The average value of optical density was 5.10 +/- 0.32, 3.52 +/- 0.35, and 8.12 +/- 0.31, respectively. Hsp70 was localized in nucleus and cellular cytoplasm while Grp94 and IgG were mainly localized in cellular cytoplasm. Ten cases showed Hsp70 was co-localized with IgG and Eighteen cases showed Grp94 was co-localized with IgG. CONCLUSION: High expression of Hsp70, Grp94 and IgG in Human lung carcinomas suggested that Hsp70, Grp94 and IgG might play an important role in the development of human lung carcinoma. IgG is co-localized with Hsp70 or Grp94. The study will lay a theorical basis for further research on anti-tumor immunotherapy.

[The changes of stem cells, immune cells and cytokines in peripheral blood after marrow stem cell mobilization in macaqne].

AIM: To explore the changes of white blood cells(WBC), neutrophils, stem cells, immune cells and cytokines after marrow stem cell mobilization by GM-CSF in macaqne. METHODS: Macaqne were injected with GM-CSF 8 mug/(kg.d) for 5 days. The ratio, number and subpopulation of WBC were observed by Blood Cell Autoanalyzer; the ratios of CD133(+), CD34(+), CD3(+), CD4(+), CD8(+), CD56(+) cells were identified by FCM; and levels of TNF-alpha, IL-1beta, IL-2 were tested by ELISA on 0, 2, 4, 6, 8, 10 day after stem cell mobiliation by GM-CSF. RESULTS: The ratios and number of CD133(+) cells, CD34(+) cells, WBC and neutrophils were significantly increased (P<0.01). The number of above cells was elevated to 6.4, 9.1, 117 and 163.3 times of normal at 6 day after stem cell were mobilized, but the ratios of CD133(+) cells and CD34(+) cell were decreased to the normal level after stem cell mobilization stopped while that of WBC and neutrocytes kept increasing over 8 d . The ratios of CD3(+), CD4(+), CD8(+), CD16(+) were decreased during 1 to 6 d, but came to the normal level at 8 d and increased at 10 d after stem cell were mobilized and the number of those cells in blood increased to 4.1, 4.0, 2.9 and 4.3 times when compared with the normal level. The concentration of TNF-alpha, IL-1beta, IL-2 in peripheral blood was significantly increased and IL-2 level was increased higher and longer than that of TNF-alpha and IL-1beta. CONCLUSION: CD133(+), CD34(+) cells and WBC, neutrophils in peripheral blood are greatly increased after GM-CSF mobilization, but WBC and neutrophils keep increasing longer than that of CD133(+), CD34(+) cells. The results of the number of CD3(+), CD4(+), CD8(+), CD16(+) cells and the levels of TNF-alpha, IL-1beta, IL-2 in peripheral blood indicate that marrow stem cell mobilization by GM-CSF can also stimulate immune cell proliferation and differentiation while increasing cytokine producing.

[A method to avoid the fixator failure by using pedicle screw combined vertebroplasty for spine fractures].

OBJECTIVE: To study a new implant material (carbonated hydroxyapatite, CHA) united pedicle screw to cure spine fracture. METHODS: Thirty-two cases of spine compressed fracture were used with pedicle screw fixator and vertebroplasty. Before operation, patients' vertebral body were compressed (46 + 21)% (20% approximately 70%) on average. In operation, broken vertebral body was reposition through pedicle screw technique, then used self-made syringe to inject CHA into anterior and central column of broken vertebral body through pedicle. And all of patients were not given any bone-graft. RESULTS: In 6 - 26 months followed-up, no immunologic rejection was found about hydroxyapatite, and no any broken of the screws and shafts was found, no loosing and other complications either. All the patients could move in 3 - 5 days after operation. The height of the broken vertebral body were reduced 97% compared with pre-operation. And CHA in vertebral body was degraded gradually, and at the same time it was replace by new bone in vertebral body. After operation, VAS score was 61 +/- 32, and there was significant difference compared with pre-operation. CONCLUSIONS: The pedicle screw fixation united vertebroplasty is an efficient way to prevent the failure of the treatment of spine fracture.