Comprehensive profiling and characterization of the absorbed components and metabolites in mice serum and tissues following oral administration of Qing-Fei-Pai-Du decoction by UHPLC-Q-Exactive-Orbitrap HRMS.

Qing-Fei-Pai-Du decoction (QFPDD) is a Chinese medicine compound formula recommended for combating corona virus disease 2019 (COVID-19) by National Health Commission of the People's Republic of China. The latest clinical study showed that early treatment with QFPDD was associated with favorable outcomes for patient recovery, viral shedding, hospital stay, and course of the disease. However, the effective constituents of QFPDD remain unclear. In this study, an UHPLC-Q-Orbitrap HRMS based method was developed to identify the chemical constituents in QFPDD and the absorbed prototypes as well as the metabolites in mice serum and tissues following oral administration of QFPDD. A total of 405 chemicals, including 40 kinds of alkaloids, 162 kinds of flavonoids, 44 kinds of organic acids, 71 kinds of triterpene saponins and 88 kinds of other compounds in the water extract of QFPDD were tentatively identified via comparison with the retention times and MS/MS spectra of the standards or refereed by literature. With the help of the standards and in vitro metabolites, 195 chemical components (including 104 prototypes and 91 metabolites) were identified in mice serum after oral administration of QFPDD. In addition, 165, 177, 112, 120, 44, 53 constituents were identified in the lung, liver, heart, kidney, brain, and spleen of QFPDD-treated mice, respectively. These findings provided key information and guidance for further investigation on the pharmacologically active substances and clinical applications of QFPDD.

["SHAO's five-needle method" as the main treatment for allergic rhinitis and asthma syndrome: a multi-center randomized controlled trial].

OBJECTIVE: To compare the clinical effect differences between "SHAO's five-needle method" and routine acupoint selection on allergic rhinitis and asthma syndrome. METHODS: A total of 210 patients with allergic rhinitis and asthma syndrome were randomly divided into an observation group (105 cases, 4 cases dropped off) and a control group (105 cases, 4 cases dropped off). The patients in the observation group were treated with "SHAO's five-needling method", and the acupoints of Feishu (BL 13), Dazhui (GV 14), Fengmen (BL 12), Yintang (GV 29), Shangyingxiang (EX-HN 8) and Hegu (LI 4), etc. were selected; the patients in the control group was treated with routine acupuncture, and the acupoints of Feishu (BL 13), Zhongfu (LU 1), Taiyuan (LU 9), Dingchuan (EX-B 1), Danzhong (CV 17), Yintang (GV 29), Fengmen (BL 12) and Zusanli (ST 36), etc. were selected. The treatment in the two groups was given once a day, 6 times a week, for 4 weeks. The score of symptoms and signs was observed before and after treatment as well as 1 month, 2 months and 3 months after treatment. The forced expiratory volume in 1 second (FEV1), peak expiratory flow (PEF) and eosinophils in peripheral blood were measured before and after treatment in the two groups. After treatment, the clinical therapeutic effect was compared between the two groups. RESULTS: The total effective rate was 98.0% (99/101) in the observation group, which was superior to 94.1% (95/101) in the control group (P<0.01). Compared before treatment, the total score of symptoms and signs in the two groups was significantly decreased at 1, 2, 3 and 4 weeks of treatment (P<0.01); after treatment and at each time point of follow-up, the total score of symptoms and signs in the observation group was lower than that in the control group (P<0.01). Compared with 4 weeks of treatment, the total score of symptoms and signs at each time point of follow-up was not statistically different in the observation group (P>0.05), and the total score of symptoms and signs in the third month of follow-up in the control group was significantly increased (P<0.05). After treatment, FEV1 and PEF in the two groups were increased (P<0.01), eosinophil count in peripheral blood was decreased (P<0.01), and the improvement in the observation group was greater than that in the control group (P<0.01, P<0.05). CONCLUSION: "SHAO's five-needle method" can improve the clinical symptoms and pulmonary function, reduce the count of eosinophils in peripheral blood in patients with allergic rhinitis and asthma syndrome, and the curative effect is better than routine acupuncture.

Mitochondrial aconitase controls adipogenesis through mediation of cellular ATP production.

Mitochondrial aconitase (Aco2) catalyzes the conversion of citrate to isocitrate in the TCA cycle, which produces NADH and FADH2, driving synthesis of ATP through OXPHOS. In this study, to explore the relationship between adipogenesis and mitochondrial energy metabolism, we hypothesize that Aco2 may play a key role in the lipid synthesis. Here, we show that overexpression of Aco2 in 3T3-L1 cells significantly increased lipogenesis and adipogenesis, accompanied by elevated mitochondrial biogenesis and ATP production. However, when ATP is depleted by rotenone, an inhibitor of the respiratory chain, the promotive role of Aco2 in adipogenesis is abolished. In contrast to Aco2 overexpression, deficiency of Aco2 markedly reduced lipogenesis and adipogenesis, along with the decreased mitochondrial biogenesis and ATP production. Supplementation of isocitrate efficiently rescued the inhibitory effect of Aco2 deficiency. Similarly, the restorative effect of isocitrate was abolished in the presence of rotenone. Together, these results show that Aco2 sustains normal adipogenesis through mediating ATP production, revealing a potential mechanistic link between TCA cycle enzyme and lipid synthesis. Our work suggest that regulation of adipose tissue mitochondria function may be a potential way for combating abnormal adipogenesis related diseases such as obesity and lipodystrophy.

A Potential Nutraceutical Candidate Lactucin Inhibits Adipogenesis through Downregulation of JAK2/STAT3 Signaling Pathway-Mediated Mitotic Clonal Expansion.

The prevalence of obesity has increased dramatically worldwide in the past ~50 years. Searching for safe and effective anti-obesity strategies are urgently needed. Lactucin, a plant-derived natural small molecule, is known for anti-malaria and anti-hyperalgesia. The study is to investigate whether lactucin plays a key role in adipogenesis. To this end, in vivo male C57BL/6 mice fed a high-fat diet (HFD) were treated with 20 mg/kg/day of lactucin or vehicle by gavage for seven weeks. Compared with vehicle-treated controls, Lactucin-treated mice showed lower body mass and mass of adipose tissue. Consistently, in vitro 3T3-L1 cells were treated with 20 µM of lactucin. Compared to controls, lactucin-treated cells showed significantly less lipid accumulation during adipocyte differentiation and lower levels of lipid synthesis markers. Mechanistically, we showed the anti-adipogenic property of lactucin was largely limited to the early stage of adipogenesis. Lactucin-treated cells fail to undergo mitotic clonal expansion (MCE). Further studies demonstrate that lactucin-induced MCE arrests might result from reduced phosphorylation of JAK2 and STAT3. We then asked whether activation of JAK2/STAT3 would restore the inhibitory effect of lactucin on adipogenesis with pharmacological STAT3 activator colivelin. Our results revealed similar levels of lipid accumulation between lactucin-treated cells and controls in the presence of colivelin, indicating that inactivation of STAT3 is the limiting factor for the anti-adipogenesis of lactucin in these cells. Together, our results provide the indication that lactucin exerts an anti-adipogenesis effect, which may open new therapeutic options for obesity.

[Effect of zige lyophilized powder for injection in improving acute cerebral microcirculation disturbance in rats].

OBJECTIVE: To investigate the effect of Zige lyophilized powder for injection in improving the acute cerebral microcirculation disturbance in rats. METHOD: Window craniotomy was performed for rats after the drug administration for 14 days. The experimental microcirculation disturbance model was duplicated with high molecule dextran. After the drug administration, the micro-vein diameters of cerebral pla mater of various groups were observed and recorded under the biological microscope. The blood flow volume was monitored by laser Doppler flow-meter. HCT was measured by the electric resistance method. The hemorheological indexes were detected by the auto-hemorheological instrument. RESULT: Zige lyophilized powder for injection (16.40, 32.70, 65.40 mg x kg(-1)) could significantly expand the micro-vein diameter of cerebral pla mater, improve the downward trend of the blood flow volume, and reduce the various hemorheological indexes. CONCLUSION: Zige lyophilized powder for injection shows the effect in improving the cerebral microcirculation.

Effect of Morinda officinalis capsule on osteoporosis in ovariectomized rats.

AIM: To explore the therapeutic effects of Morinda officinalis capsules (MOP) on osteoporosis in ovariectomized rats. METHODS: Six-month-old female Sprague-Dawley rats were induced for postmenopausal osteoporosis (PMOP) by bilateral ovariectomy and divided into seven groups as follows: sham-operated group, ovariectomized (OVX) control group, OVX treated with xianlinggubao (XLGB) (270 mg·kg⁻¹·d⁻¹), OVX treated with alendronate sodium (ALN) (3 mg·kg⁻¹·d⁻¹), and OVX treated with Morinda officinalis capsule (MOP) of graded doses (90, 270 and 810 mg·kg⁻¹·d⁻¹) groups. Oral treatments were administered daily on the 4(th) week after ovariectomy and lasted for 12 weeks. The bone mineral density was evaluated by dual-energy X-ray absorptiometry. The tartrate-resistant acid phosphatase (TRAP), alkaline phosphatase (AKP), and osteocalcin (OC) levels in the serum and plasma were determined by standard colorimetric and enzyme immunoassays methods. Bone biomechanical properties and morphological parameters were analyzed by three-point bending test and histomorphometry respectively. RESULTS: Morinda officinalis capsules at all doses were able to significantly prevent the OVX-induced loss of bone mass due to diminishing serum AKP and TRAP levels while elevating OC level in the plasma. Morinda officinalis capsules also enhanced the bone strength and prevented the deterioration of trabecular microarchitecture. CONCLUSION: Morinda officinalis capsules possess potent anti-osteoporotic activity in OVX rats which could be an effective treatment for postmenopausal osteoporosis.

[Cell membrane chromatography and its application progress in studies of traditional Chinese medicines].

As a novel bioaffinity chromatography technique, cell membrane chromatography (CMC) was first established by Professor He in 1996, with which combined high performance liquid chromatography, cytobiology, and receptor pharmacology. The cell membrane stationary phase (CMSP) consists of porous silica coated with active cell membranes. By immersing silica into a suspension of cell membranes, the whole surface of silica was covered by the cell membranes. In CMC, the interaction of drugs or compounds with the immobilized cell membrane or its receptors is investigated using liquid chromatography. In general, with the aim to provide scientific foundation for further development and application, this paper mainly focuses on the characteristics of the cell membrane stationary phase (CMSP), the CMC analytical system, and its applications in traditional Chinese medicines (TCMs) about CMC. With the development of CMC, the breakthrough progress of it in studying active components of TCMs field is expectant.

[Effects of cycloartocarpin A and artocarpin extracted from Fructus Artocarpin Heterophylli on apoptosis of SMMC-7721 and SGC-7901 cells].

OBJECTIVE: To investigate the effects of cycloartocarpin A (ACR-2) and artocarpin (ACR-3), monomeric compounds isolated from Fructus Artocarpi Heterophylli, on apoptosis of SMMC-7721 and SGC-7901 cell lines. METHODS: SMMC-7721 and SGC-7901 cells were routinely cultured, and divided into experiment group and control group. The SMMC-7721 cells were treated with different concentrations of ACR-2 (3.46 x 10(-3), 13.82 x (-3), 55.30 x 10(-3) mmol/L) and ACR-3 (6.88 x 10(-3), 27.52 x 10(-3), 110.09 x 10(-3) mmol/L), and the SGC-7901 cells were also treated with different concentrations of ACR-2 (8.06 x 10(-3), 32.26 x 10(-3), 129.03 x 10(-3) mmol/L) and ACR-3 (2.87 x 10(-3), 11.47 x 10(-3), 45.87 x 10(-3) mmol/L), with PBS (DMSO<0.1%) as control treatment. Cell apoptosis was measured by double labeled staining with Hoechst33342/propidium iodide (PI) and TdT-mediated dUTP-biotin nick end labeling (TUNEL) and flow cytometry. RESULTS: ACR-2 and ACR-3 could induce apoptosis of SMMC-7721 and SGC-7901 cells. Some of SMMC-7721 and SGC-7901 cells demonstrated typical apoptosis after being treated with ACR-2 and ACR-3. Hoechst33342/PI staining showed that cells were fraught with overlapping nuclei and nuclear debris or lobule, and the nuclear appeared light blue. TUNEL showed that cells permeated with overlapping nuclei and nuclear debris or lobule, and the nuclear appeared brown. Less apoptotic cells were observed in negative control group, and the nuclear appeared light blue. The apoptosis rates of SMMC-7721 and SGC-7901 cells in the ACR-2 and ACR-3 treated groups were significant higher than those in the control group (P<0.05, P<0.01). CONCLUSION: ACR-2 and ACR-3 can induce apoptosis of SMMC-7721 and SGC-7901 cells.

[Effects of three compounds extracted from Tripterygium wilfordii Hook on angiogenesis in chick chorioallantoic membrane].

OBJECTIVE: To investigate the effects of three compounds extracted from Tripterygium wilfordii Hook (TW) on angiogenesis in the chick chorioallantoic membrane (CAM). METHODS: Fifty fresh Hongkong Mahua chicken eggs were divided into five groups: PBS-treated group, TW1-, TW2- and TW3-treated groups and Rg3-treated group. After disinfection, the eggs were incubated for six days in a constant temperature box with the temperature being controlled within 37.8 degrees C, then exposed CAM, laid the filter papers with specimen on the CAM, and the eggs were incubated for another two days. CAM was fixed with the mixture of methyl alcohol and acetone at room temperature for about 15 min, and then cutting the CAM, taking photos and observing the angiogenesis in the CAM. RESULTS: There were many CAM vessels in the PBS-treated group and the blood vessel net could be seen clearly. The number of CAM vessels in the TW1-, TW2- and TW3-treated groups (10 microg/egg) was much less than that in the PBS-treated group. Furthermore, the frame of the vessels was not clear, and the color was obscure. Inhibition rates of angiogenesis in the TW1-, TW2- and TW3-treated groups were 80%, 60% and 100% respectively, while the inhibition rate of angiogenesis in the Rg3-treated group (10 microg/egg) was only 10%. CONCLUSION: TW1, TW2 and TW3 can obviously restrain the angiogenesis in CAM and still need further study.