Diagnosis and treatment of novel coronavirus pneumonia based on the theory of traditional Chinese medicine.

Since the outbreak of novel coronavirus pneumonia (coronavirus disease 2019, COVID-19), it has rapidly spread to 187 countries, causing serious harm to the health of people and a huge social burden. However, currently, drugs specifically approved for clinical use are not available, except for vaccines against COVID-19 that are being evaluated. Traditional Chinese medicine (TCM) is capable of performing syndrome differentiation and treatment according to the clinical manifestations of patients, and has a better ability of epidemic prevention and control. The authors comprehensively analyzed the etiology and pathogenesis of COVID-19 based on the theory of TCM, and discussed its syndrome differentiation, treatment and prevention measures so as to provide strategies and reference for the prevention and treatment with TCM.

Dietary yeast hydrolysate and brewer's yeast supplementation could enhance growth performance, innate immunity capacity and ammonia nitrogen stress resistance ability of Pacific white shrimp (Litopenaeus vannamei).

An 8-week feeding trial was conducted to evaluate the effects of dietary yeast hydrolysate and brewer's yeast supplementation on growth, immune-related genes expression and ammonia nitrogen stress resistance of Pacific white shrimp (Litopenaeus vannamei). Three isonitrogenous and isolipidic practical diets were formulated to contain 0% (control diet), 1% yeast hydrolysate and 1% brewer's yeast, respectively. 360 juvenile L. vannamei with an initial weight (0.88 ±â€¯0.01 g) was randomly divided into 3 treatments in four replicates (30 shrimp per replicate). The results indicated that shrimp fed the diet containing 1% yeast hydrolysate had a significantly higher weight gain (WG), and specific growth rate (SGR) than that fed the control diet, and the lowest feed conversion ratio (FCR) was occurred in the 1% yeast hydrolysate supplementation group. Proximate composition in whole body and muscle among all treatments was not significantly influenced by the dietary yeast hydrolysate or brewer's yeast supplementation. The challenge test with ammonia nitrogen showed that lower cumulative survival was observed in those fed the control diet, and the highest cumulative survival was occurred at shrimp fed the 1% yeast hydrolysate supplementation. Shrimp fed the control diet had higher inflammation-related genes expression levels of tnf-α and il-1ß in the intestine than those fed the diets supplemented with 1% yeast hydrolysate or 1% brewer's yeast, however, there was no significant difference in expression level of alp in intestine among all treatments. The relative expression levels of mTOR signal pathway genes (eif4ebp, eif4e1a, eif4e2 and p70s6k) were significantly up-regulated in the shrimp fed the diets supplemented with 1% yeast hydrolysate, and the lowest gene expression levels of eif4ebp, eif4e1a, eif4e2 and p70s6k in the intestine were occurred at the control diet. The highest expression levels of the immune-related genes (dorsal, relish, and proPO) in the intestine were observed at shrimp fed the 1% yeast hydrolysate supplementation, and the lowest expression levels of these genes were occurred at shrimp fed the control diet, however, there was no significant difference in gene expression of lysozyme among all treatments. The expression levels of penaeidin3a, crustin, proPO, and IMD in the hepatopancreas were significantly influenced by the dietary yeast hydrolysate, brewer's yeast or no yeast product supplementation, shrimp fed the 1% yeast hydrolysate supplementation had higher expression levels of these genes than those fed the control diet. The present study indicated that dietary 1% yeast hydrolysate or brewer's yeast supplementation could improve growth performance, enhance innate immunity, and strengthen resistance of ammonia nitrogen stress, and dietary 1% yeast hydrolysate supplementation provides better immunostimulatory effects than brewer's yeast of L. vannamei.

Lariciresinol induces apoptosis in HepG2 cells via mitochondrial-mediated apoptosis pathway.

Lariciresinol (LA) is one of the main active ingredients in many traditional medicinal plants such as Patrinia, and has the role of anti-liver cancer. However, the precise mechanisms are unclear. This study investigated the molecular mechanisms of LA against HepG2 cells. LA anti-tumor activity was assessed with the CCK-8, Ki-67, and immunofluorescence staining. Cells apoptotic ratio was evaluated by Annexin V/PI double-staining assay. A proteomic approach was used to identify differentially expressed proteins after LA treatment. JC-1 staining was carried out to detect the mitochondrial membrane potential (ΔΨm), and the Western blot analysis was used to analyse the apoptosis-associated proteins. Our results suggested that LA significantly suppressed the viability of HepG2 cells. The CCK-8 and Ki-67 expression indicated dose-dependent decreases in cell proliferation. Flow cytometry analysis showed that LA exhibited a apoptosis-inducing effect. The proteomic study observed the presence of apoptosis-associated proteins and mitochondrial dysfunction in HepG2 cells after LA-treatment. Further analysis showed that LA could trigger the mitochondrial-mediated apoptosis pathway, based on a decrease in ΔΨm; deliver of cytochrome c; activation of caspase-9/-3 and poly(ADP-ribose) polymerase; and decrease of the proportion of Bcl-2/Bax. Collectively, our studies found that LA exhibits significant cytotoxic effects by inhibiting cell proliferation, inducing apoptosis, possibly via activation of the mitochondrial-mediated apoptosis pathway.

Synergistic cytotoxicity of ampelopsin sodium and carboplatin in human non-small cell lung cancer cell line SPC-A1 by G1 cell cycle arrested.

OBJECTIVE: To evaluate the cytotoxic effects of ampelopsin sodium (Amp-Na) and carboplatin (CBP) used alone or in combination on human non-small cell lung cancer (NSCLC) cells SPC-A1 in vitro and its related mechanism. METHODS: Cytotoxic effects were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. The synergistic effects of the drugs were calculated with coefficient of drug interaction (CDI). Cell cycle was determined by flow cytometry (FCM). The levels of p53, p21, cyclinE, cyclinD1, and phosphorylated cyclin-dependent kinase-2 (p-CDK2) were evaluated by Western blot. RESULTS: Amp-Na (6.25-200 µg/mL) and CBP (3.13-100 µg/mL) alone exhibited prominent cytotoxic activity in a concentration-dependent manner on SPC-A1 cells with 50% inhibitive concentration values of 57.07±14.46 and 34.97±6.30 µg/mL, respectively. Drug combinations were associated with significantly higher cytotoxic effects than each drug alone (P<0.05 or 0.01). The CDI analysis confirmed the synergy of Amp-Na and CBP on inhibiting cancer cell viability across a wide concentration range (CDI <1). FCM and Western blot showed that synergistic cytotoxic effects of Amp-Na and CBP were related to G1 arrested which mainlym ediated by p 21 through the inhibition of CDK2 activity independent of the p53 tumor suppressor pathway. CONCLUSIONS: Amp-Na exhibits anticancer activities and enhances the antitumor activities of CBP through up-regulation of p21 and inhibition of CDK2 activity in human NSCLC cells SPC-A1. These results suggest that Amp-Na may be applied to enhance the anticancer action of CBP.

Proteomic analysis of apoptosis induction by lariciresinol in human HepG2 cells.

Lariciresinol (LA) is a traditional Chinese medicine possessing anticancer activity, but its mechanism of action remains unclear. The present study explored the effects of LA on human HepG2 cells and the underlying mechanism. Our data indicated that LA inhibited cell proliferation and induced cell cycle arrest in S phase, subsequently resulting in apoptosis in HepG2 cells. Using a proteomics approach, eight differentially expressed proteins were identified. Among them, three proteins, glyceraldehyde-3-phosphate, UDP-glucose 4-epimerase, and annexin A1, were upregulated, while the other five proteins, heat shock protein 27, haptoglobin, tropomodulin-2, tubulin alpha-1A chain, and brain acid soluble protein 1, were downregulated; all of these proteins are involved in cell proliferation, metabolism, cytoskeletal organization, and movement. Network analysis of these proteins suggested that the ubiquitin-conjugating enzyme (UBC) plays an important role in the mechanism of LA. Western blotting confirmed downregulation of heat shock protein 27 and upregulation of ubiquitin and UBC expression levels in LA-treated cells, consistent with the results of two-dimensional electrophoresis and a STRING software-based analysis. Overall, LA is a multi-target compound with anti-cancer effects potentially related to the ubiquitin-proteasome pathway. This study will increase our understanding of the anticancer mechanisms of LA.

[Study on the effect of grape seed proanthocyanidins on increasing the radiosensitivity for X-ray].

OBJECTIVE: To investigate the effect of Grape Seed Proanthocyanidins (GSPs) on enhancing the radiosensitivity of human hepatic carcinoma cell line HepG2, human cervical cancer cell line Hela and human leukemia cell line K562 for X-ray in vitro. METHODS: The killing effect of GSPs combined with X-ray on cells was evaluated by SRB and clone formation assay. RESULTS: The GSPs had obvious cytotoxicity on three cell lines in a dose-dependent and time-dependent manners. However, inhibition rate of different cell line were quite different, the strongest one was human leukemia K562 cells and the others were weak. The sensitization ranges calculated by univariate analysis were 6.25-12.5 microg/mL for human leukemia K562 cells. Sensitization enhancement ratio was 1.94 using curve fitting method for K562 cells. CONCLUSION: GSPs can obviously enhance the radiosensitivity of cancer cells in vitro. The mechanism of sensitization effect may be related to the effects of GSPs on oxygen balance and cell cycle.

[Effects of extracts from Patrinia heterophylla on gene expression patterns during morphogenesis of chicken limb buds in vivo].

OBJECTIVE: To study the effects of the extracts from Patrinia heterophylla on gene expression patterns during morphogenesis of chicken limb buds in vivo. METHODS: Implanted a bead into an chicken embryo, which was soaked in the extracts from Patrinia heterophylla. Detected the extracts-induced morphogenesis changes (Myf5, Myod and PCNA). RESULTS: The extracts from Patrinia heterophylla (200 mg/mL) could affect limb bud development, reduce gene expression of MyfS, MyoD and PCNA. CONCLUSION: The extracts from Patrinia heterophylla can inhibit cell differentiation and proliferation.

[An experimental observation of pulmonary fibrosis in rats and its transforming growth factor beta1 treated by total hedysarum polybotyssaccharide].

OBJECTIVE: To treat pulmonary fibrosis and study its mechanisms. METHODS: Choosing randomly 24 Wistar rats with normal sodium intratracheal injection as normal control group, the rests with heomycin A5 induced fibrosis were divided randomly into model group 24 as positive control, prednisone group 24, total hedysarum polybotyssaccharide (THPS) group 24, THPS and small dose prednisone group 24, and treated with different drugs. 6 rats of every group were put to death and observed pathological section, using imaging processing computer to quantitative analysis histomorphology, collagen, and transforming growth beta1, (TGF-beta1) on 7, 14, 30, 60 days. RESULTS: The group treated by THPS combination small dose prednisone showed up the most effects in the 3 treatment groups. CONCLUSION: THPS combination small dose prednisone to treat pulmonary fibrosis of rats is better than classic ways and its efficacy of inhibition TGF-beta1 may be a mechanism.

[Effect of Patrinia scabra extracts on the level of serum cytokine in sarcoma 180 ascitic tumor burdened mice].

OBJECTIVE: To explore the effect of Potrinia scabro extracts (PSE) on the level of serum cytokine in Sarcoma 180 ascitic tumor burdened mice and its mechanism of anti-tumor. METHODS: The mice model of Sarcoma 180 ascitic tumor were established and divided into five groups randomly, including the model group with normal saline solution, the positive group with 10 mg/kg cytoxan and PSE treated groups at doses of 2.0 g/kg, 1.0 g/kg, 0.5 g/kg intraperitoneally for 10 days. The level of serum cytokine Th1 (IL-2, IL-12, IFN-gamma, IFN-alpha) and Th2 (IL-6, IL-10) were measured by double antibody sandwich ELISA assay. RESULTS: Compared with model group of Sarcoma 180 ascitic tumor burdened mice,the level of IL-2 and IFN-gamma increased in PSE 2.0 g/kg group, but the IL-6 and IL-10 decreased in PSE 2.0 g/kg and 1.0 mg/kg groups. CONCLUSION: PSE has anti-tumor effect in vivo that could be related to the level variation of IL-2, IFN-gamma, IL-6 and IL-10 in tumor burdened mice.

[Effects of Patrinia scabra bunge macroporous adsorptive resins extracts on erythrocyte immune function in tumor bearing mice].

OBJECTIVE: To study the erythrocyte immuno-regulatory effect of Patrinia scabra Bunge extracts extracted by macroporous adsorptive resins in tumor bearing mice. METHODS: Patrinia scabra Bunge was extracted by macroporous adsorptive resins, and the amount of polysaccharides and saponins in the extract were determined. Mice bearing S180 tumor were treated with the extract and their survival prolongation rate, erythrocyte rosette formation rates of C3b receptor (ERR-CR), immune complex (ERR-IC) and tumor cell (ERR-TC), as well as the CD35 and CD44s were observed. RESULTS: Polysaccharide content was 21.4%, saponin 41.8% in the extract. As compared with the model group, the survival rate was increased, the erythrocyte immune function was improved (showed increase of ERR-CR and ERR-TC, decrease of ERR-IC), and the amount of CD35 and CD44s in red blood cell membrane increased in mice after being treated with the extract (P < 0.05 or P < 0.01). CONCLUSION: Extract of Patrinia scabra Bunge extracted by macroporous adsorptive resins can regulate the erythrocyte immune function to a certain extent.

[Studies on extraction process optimization of patrinia scabra Bunge saponins].

OBJECTIVE: To optimize the conditions for the extraction of Patrinia scabra Bunge saponins. METHODS: Orthogonal experimental design and ultrasonic method were employed to examine the conditions for the extraction by determination of saponins. RESULTS: The optimun condition for the extraction of Patrinia scabra Bunge saponins was as follows: 65% ethanol for 40 minutes, 55 degrees C and 210 watt of ultrasonic efficinecy. CONCLUSION: The extraction method of Patrinia scabra Bunge sponins is simple and efficient.

Effects of body-resistance strengthening and tumor-suppressing granules on immune adhesion function of red blood cells and expression of metastasis protein CD44 in tumor cells of patients with esophageal carcinoma.

AIM: To investigate the effects of Fuzheng Yiliu granules (body-resistance strengthening and tumor-suppressing granules) in patients with esophageal carcinoma. METHODS: We compared the immune adherent properties of red blood cells (RBCs), the expression of metastasis protein CD44, and the metastasis inhibition factor nm23, in esophageal carcinoma tumor cells of patients before and after radiotherapy in the presence and absence of orally administered Fuzheng Yiliu granules. Sixty-three hospitalized patients with esophageal carcinoma were treated with standard radiotherapy and randomly divided into treatment group (n = 30) treated with both radiotherapy and Fuzheng Yiliu granules and control group (n = 33) given radiotherapy only. Blood samples and tumor tissue were obtained before and after 21 d of treatment. The rosette rates for complement receptor type 3b (C3bRR) and immune complex receptor (ICRR) on RBCs were measured by erythrocyte immunological methods. Expression of CD44 and nm23 in tumor tissue sections was determined by immunohistochemical staining with monoclonal antibodies CD44v6 ad nm23H-1, respectively. RESULTS: The positivity of RBC-C3bRR before and after 21 d of treatment increased from 7.78% +/- 1.59% to 10.03% +/- 2.01% in the double treatment group, while it changed only slightly from 7.18% +/- 1.29% to 7.46% +/- 1.12% in the radiotherapy group. The positive rate for RBC-ICRR decreased from 37.68% +/- 2.51% to 22.55% +/- 1.65% after the double treatment, and from 37.28% +/- 2.41% to 24.69% +/- 1.91% in radiotherapy group at the same time points. The difference in erythrocyte immune adherent function between the two groups was significant (P < 0.01, t-test). The CD44(+)-cases were reduced from 21 (70.00%) to 12 (40.00%) after treatment with Fuzheng Yiliu granules, whereas the CD44(+)-cases (69.70%) in the radiotherapy group remained unchanged. The difference between the treatment (40.00%) and control (69.70%) groups was significant (P < 0.05). Although the nm23(+)-cases were increased from 4 (13.33%) to 6 (20.00%) in the double treatment group and from 6 (18.18%) to 7 (21.21%) in the radiotherapy group, the difference was not significant (P > 0.05). CONCLUSION: Fuzheng Yiliu granules enhance the immune adhesion function of RBCs and reduce the number of CD44(+)-cells in esophageal carcinoma patients, suggesting a potential role of these Chinese herbals in suppression of invasion and metastasis of malignant cells. However, this anti-metastatic effect has yet to be validated in vivo.

[Effect of Fuzheng Yiliu Granule contained-serum from tumor bearing mice on apoptotic rate, free radicals content and mitochondrial membrane potential of hepatoma cell lines H22 in vitro].

OBJECTIVE: To observe the effect of traditional Chinese herbs Fuzheng Yiliu Granule (FYG)-contained serum from tumor bearing mice on apoptotic rate, free radicals content and mitochondrial membrane potential of hepatoma cell lines H22 in vitro. METHODS: The effect of FYG drug-serum on apoptosis of hepatoma cell line H22 was determined using flow cytometry. The changes of DNA RNA, free radicals and mitochondrial membrane potential (delta psi m) in H22 cell were detected through laser scanning confocal microscope. RESULTS: FYG-contained serum can induce the apoptosis of H22 cell, enhance the free radicals content, and reduce the content of DNA RNA and delta psi m of H22 cell in vitro. CONCLUSION: The apoptosis of hepatoma cell line H22 induced by FYG is probably correlated to the change of free radicals content and delta psi m.

[Study on depressant effects of THPS on S180 tumor-bearing-mice and its mechanisms].

OBJECTIVE: to study depressant effect of total hedysarum polybotys saccharids (THPS) on S180 tumor-bearing-mice and its mechanisms. METHODS: THPS was extracted from Radix Xedysari with water and precipitated with ethanol, determining its molecular weight, purity, saccharide and aldonic acid content. 90 Kunming mice were divided into 9 groups randomly. One group was the normal group, the others were divided into 8 groups randomly after inculating S180 tumors and were treated with THPS and THPS combination cyclophosphamide (CY) in low, moderate and high dose, to put them to death after 14 days. To determine every tumor weight, the rate of depressant tumor, the contents of IL-2 and TNF-alpha with ELISA, and NF-kappaB with immunochemistry. RESULTS: The moderate dose THPS conspicuously possessed a depressant effect on S180 tumor and joint action with combination CY and increased the contents of IL-2, TNF-alpha and NF-kappaB of mice. CONCLUSION: THPS of moderate dose possesses a depressant effect on S180 tumor through increasing the contents of IL-2, TNF-alpha and NF-kappaB of mice.

[Erythrocyte immunologic function effects of Patrinia scabra extracts by macroporous adsorptive resins on mice burdened transplanted tumor].

OBJECTIVE: To research the erythrocyte immunoregulation effects of Patrinia scabra extracts by macroporous adsorptive resins on mice burdened transplanted tumor. METHODS: Extracts of Patrinia scabra Bunge were separated by macroporous adsorptive resins, ingredients were analysised. Mice burdened transplanted tumor were given extracted drugs. Life prolongation rate was observed, erythrocyte immunologic function and the CD35, CD44s contents of red blood cell were evaluated. RESULTS: Polysaccharide and saponin accounted for 8.4% and 48.4%. Extracts could porolong life expectancy of mice, improve erythrocyte immunolgic function and increase the CD35 and CD44s contents of red blood cell. CONCLUSION: Extracts of Patrinia scabra Bunge by macroporous adsorptive resins have erythrocyte immunoregulation effects on mice burdened transplanted tumor.

[Effects of fuzheng yiliu granules on apoptotic rate and mitochondrial membrane potential of hepatocellular carcinoma cell line H22 from mice].

OBJECTIVE: To evaluate the effects of Fuzheng Yiliu Granules (FZYLG) on apoptotic rate and mitochondrial membrane potential (Delta psi m) of hepatocellular carcinoma cell line H22 from mice. METHODS: Forty-eight mice inoculated with H22 cells were randomly divided into four groups: untreated group, cyclophosphamide-treated group, high-dose FZYLG-treated group and low-dose FZYLG-treated group. After 14 days of corresponding treatment, H22 cells in each group were stained with propidium iodide, and the apoptotic rates were detected by flow cytometry (FCM). The rhodamine 123 was used as a fluorescence probe to label the H22 cells, and the fluorescence intensities were observed with laser scanning confocal microscope. The fluorescence intensity of H22 cells indicated the Delta psi m of H22 cells. RESULTS: FZYLG could significantly increase the apoptotic rate while reduce the Delta psi m of H22 cells from mice as compared with those in the untreated group. CONCLUSION: The antitumor effects of FZYLR on H22 cells from mice are related to decreasing the Delta psi m and then inducing the apoptosis of the H22 cells.