Hesperitin-Copper(II) Complex Regulates the NLRP3 Pathway and Attenuates Hyperuricemia and Renal Inflammation.

BACKGROUND: Hyperuricaemia (HUA) is a disorder of purine metabolism in the body. We previously synthesized a hesperitin (Hsp)-Cu(II) complex and found that the complex possessed strong uric acid (UA)-reducing activity in vitro. In this study we further explored the complex's UA-lowering and nephroprotective effects in vivo. METHODS: A mouse with HUA was used to investigate the complex's hypouricemic and nephroprotective effects via biochemical analysis, RT-PCR, and Western blot. RESULTS: Hsp-Cu(II) complex markedly decreased the serum UA level and restored kidney tissue damage to normal in HUA mice. Meanwhile, the complex inhibited liver adenosine deaminase (ADA) and xanthine oxidase (XO) activities to reduce UA synthesis and modulated the protein expression of urate transporters to promote UA excretion. Hsp-Cu(II) treatment significantly suppressed oxidative stress and inflammatory in the kidney, reduced the contents of cytokines and inhibited the activation of the nucleotide-binding oligomerization domain (NOD)-like receptor thermal protein domain associated protein 3 (NLRP3) inflammatory pathway. CONCLUSIONS: Hsp-Cu(II) complex reduced serum UA and protected kidneys from renal inflammatory damage and oxidative stress by modulating the NLRP3 pathway. Hsp-Cu(II) complex may be a promising dietary supplement or nutraceutical for the therapy of hyperuricemia.

Acupuncture promotes tear secretion by up-regulating VIP/cAMP/PKA/AQP5 signaling in guinea pigs with aqueous tear deficiency dry eye. / 基于VIP/cAMP/PKA/AQP5信号通路探讨针刺治疗水液缺乏型干眼的作用机制.

OBJECTIVES: To observe the effect of acupuncture on the ocular surface symptoms and the protein expression of vasoactive intestinal peptide (VIP) / cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) / aquaporin 5(AQP5) signaling pathway in lacrimal gland tissue of aqueous tear deficiency (ATD) type dry eye model, so as to investigate its mechanism underlying improvement of ATD. METHODS: British shorthair guinea pigs were randomly divided into blank control, model, acupuncture, sham-acupuncture and medication group, with 8 guinea pigs in each group. The ATD model was established by subcutaneous injection of scopolamine hydrobromide (0.6 mg/dose, 4 times/d for 10 days). For guinea pigs of the acupuncture group, filiform needles were inserted into bilateral "Jingming"(BL1), "Cuanzhu"(BL2), "Sizhukong"(TE23), "Taiyang"(EX-HN5), and "Tongziliao"(GB1) for 15 min. For guinea pigs of the sham-acupuncture group, a blunt filiform needle was used to repeatedly prick (not pierce) the skin of the same acupoints mentioned above. The treatment in the above two groups was conducted once daily for 14 days. The guinea pigs in the medication group received administration of sodium hyaluronate eye drops in both eyes, three times a day for 14 days. The objective tests of tear film break-up time (BUT), corneal fluorescein staining score (FLS) and phenol red thread (PRT) test were conducted before and after modeling and after the intervention. After the intervention, the lacrimal index (weight of lacrimal gland/body weight) was calculated. Histopathological changes of the lacrimal gland were observed after H.E. staining. The expression of AQP5 in the lacrimal gland were detected by immunofluorescence, and the contents of VIP and AQP5 in the lacrimal gland were measured by ELISA, the protein expression levels of VIP, cAMP, PKA, p-PKA and AQP5 in the lacrimal gland were detected by Western blot. RESULTS: In comparison with the blank control group, the PRT, BUT, lacrimal index, AQP5 immunoactivity, contents of VIP and AQP5, and protein expression levels of VIP, cAMP, PKA, p-PKA and AQP5 were significantly decreased(P<0.01, P<0.05), and FLS was obviously increased (P<0.01) in the model group . Compared to the model group, the PRT, BUT, lacrimal index, AQP5 immunoactivity, contents of VIP and AQP5, and expression levels of VIP and AQP5 in both acupuncture and medication groups, and the expression levels of cAMP, PKA, p-PKA in the acupuncture group were considerably increased (P<0.01, P<0.05), while the FLS was markedly decreased in both acupuncture and medication groups (P<0.01, P<0.05). Compared with the medication group, the acupuncture group had increased PRT (P<0.05). CONCLUSIONS: Acupuncture intervention is effective in reducing ocular surface damage and promoting tear secretion in guinea pigs with ATD, which may be related to its function in activating VIP/cAMP/PKA signaling, and promoting the expression of AQP5 in the lacrimal gland.

Formononetin improves the inflammatory response and bone destruction in knee joint lesions by regulating the NF-kB and MAPK signaling pathways.

Formononetin (FMN) is a phytoestrogen that belongs to the isoflavone family. It has antioxidant and anti-inflammatory effects, as well as, many other biological activities. Existing evidence has aroused interest in its ability to protect against osteoarthritis (OA) and promote bone remodeling. To date, research on this topic has not been thorough and many issues remain controversial. Therefore, the purpose of our study was to explore the protective effect of FMN against knee injury and clarify the possible molecular mechanisms. We found that FMN inhibited osteoclast formation induced by receptor activator of NF-κB ligand (RANKL). Inhibition of the phosphorylation and nuclear translocation of p65 in the NF-κB signaling pathway plays a role in this effect. Similarly, during the inflammatory response of primary knee cartilage cells activated by IL-1ß, FMN inhibited the NF-κB signaling pathway and the phosphorylation of the ERK and JNK proteins in the MAPK signaling pathway to suppress the inflammatory response. In addition, in vivo experiments showed that both low- and high-dose FMN had a clear protective effect against knee injury in the DMM (destabilization of the medial meniscus) model, and the therapeutic effect of high-dose FMN was stronger. In conclusion, these studies provide evidence of the protective effect of FMN against knee injury.

[Acupuncture mitigates ocular surface inflammatory response via α7nAChR/NF-κB p65 signaling in dry eye guinea pigs].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the ocular surface inflammation and α7 nicotinic acetylcholine receptor (α7nAChR) / nuclear factor kappa-B (NF-κB) p65 signal pathway in guinea pigs with dry eye, so as to explore its underlying mechanism. METHODS: A total of 32 male British tricolor short haired guinea pigs were randomized into blank control, model, EA and sham acupuncture groups, with 8 guinea pigs in each group. The dry eye model was established by subcutaneous injection of scopolamine hydrobromide solution (0.6 mg/0.2 mL each time, 4 times a day for 10 days). Guinea pigs of the EA group was treated with EA at bilateral "Cuanzhu" (BL2) and "Taiyang" (HN5), and manual acupuncture at bilateral "Jingming" (BL1), "Sizhukong" (SJ23), "Tongziliao" (GB1) for 15 min, once daily for 14 days. For animals of the sham acupuncture group, a blunt needle was used to prick the skin surface of the acupoints, the acupoint selection and stimulation time were the same as those in the EA group. Before and after modeling and after the intervention, the breakup time (BUT) of lacrimal film, sodium fluorescein coloring (Fl) state of corneal epithelium and phenol red thread (PRT) moist length were recorded for assessing the severity of dry eye. The density of activated immune cells around the corneal epithelial stromal cells was determined by corneal confocal microscopy. The contents of interleukin-4 (IL-4), IL-6, IL-10, tumor necrosis factor α (TNF-α) in the cornea and lacri-mal gland tissues were determined by ELISA, and the expression levels of α7nAChR and NF-κB p65 in the cornea and lacrimal gland were detected by immunohistochemistry and Western blot, separately. RESULTS: Compared with the blank control group, the corneal Fl, density of activated immune cells of corneal epithelium, contents of IL-6, IL-10 and TNF-α in both corneal and lacrimal gland tissues, NF-κB p65 cell positive rate and protein expression of lacrimal gland and corneal tissues were significantly increased (P<0.01, P<0.05), while the BUT, PRT and lacrimal gland α7nAChR cell positive rate considerably decreased (P<0.01) in the model group. In comparison with the model group, the level of corneal Fl, density of the activated immune cells of corneal epithelium, contents of corneal and lacrimal IL-6 and TNF-α, and corneal and lacrimal NF-κB p65 cell positive rates and protein expressions were remarkably down-regulated in the EA group (P<0.01, P<0.05), rather than in the sham acupuncture group (P>0.05) except content of corneal IL-10, lacrimal NF-κB p65 cell positive rate and lacrimal α7nAChR protein expression, whereas the levels of BUT, PRT, corneal and lacrimal IL-10 and corneal and lacrimal α7nAChR cell positive rates and protein expressions significantly up-regulated in the EA group (P<0.01, P<0.05), rather than in the sham acupuncture group (P>0.05) except corneal TNF-α and corneal NF-κB p65 protein expression. CONCLUSION: EA can improve corneal and lacrimal function in dry eye guinea pigs, which may be associated with its actions in increasing the expression of α7nAChR, inhibiting the nuclear translocation of NF-κB, and reducing the activated immune cells and inflammatory reaction.

Action mechanisms of two key xanthine oxidase inhibitors in tea polyphenols and their combined effect with allopurinol.

BACKGROUND: Tea polyphenols have been reported to have the effect of lowering uric acid. However, there are few studies on the inhibitory effects and molecular mechanisms of specific catechins on the urate-metabolizing enzyme xanthine oxidase (XO). In this research, multiple spectroscopic methods and computer simulations were used to determine the inhibitory ability and mechanisms of epigallocatechin gallate (EGCG) and gallocatechin gallate (GCG) on XO. RESULTS: Herein, EGCG and GCG reversibly inhibited XO activity in a mixed manner, with IC50 values of 40.50 ± 0.32 and 33.60 ± 0.53 µmol L-1 , and also decreased the superoxide anion radical (O2 - ) of the catalytic system by reducing the XO molecule and inhibiting the formation of uric acid. The combination of EGCG or GCG with allopurinol showed synergistic inhibition on XO. The binding of EGCG or GCG to XO with moderate affinity formed a stable complex by hydrogen bonds and van der Waals forces. The presence of EGCG and GCG made the structure of XO more stable and compact. The two inhibitors bound to the vicinity of flavin adenine dinucleotide (FAD) in XO, hindering the entry of substrate; thus the activity of XO was suppressed. CONCLUSION: Both EGCG and GCG are excellent natural XO inhibitors, and inhibited the activity of XO by occupying the channel of the substrate to enter the active center and interfering with the dual substrate reaction catalyzed by XO. These findings provide a scientific basis for the application of catechins in dietary supplements and medicines with lowering uric acid effects. © 2022 Society of Chemical Industry.

[Phosphorus forms in marsh soils with different years of Spartina alterniflora invasion in the Minjiang River estuary, China]. / é—½æ±Ÿæ²³å£äº’èŠ±ç±³è‰ä¸åŒå ¥ä¾µå¹´é™æ¹¿åœ°åœŸå£¤ç£·èµ‹å­˜å½¢æ€.

We examined the effects of Spartina alterniflora invasion on phosphorus forms of marsh soils, based on the method of space-for-time substitution by selecting S. alterniflora marshes with different invasion years (SA1, 5-6 years; SA2, 8-10 years; and SA3, 12-14 years) in Shanyutan of the Minjiang River estuary. The results showed that in marsh soils of different invasion years, the proportion of hardly decomposable phosphorus (HCl-Pi and Residual-P) was the highest (46.4%-46.7%), followed by moderately decomposable phosphorus (NaOH-Pi, NaOH-Po and Sonic-Pi) (40.0%-44.0%), and the easily decomposable phosphorus (Resin-Pi, NaHCO3-Pi and NaHCO3-Po) was the lowest (9.5%-13.3%). With increasing invasion years of S. alterniflora, soil phosphorus forms and their spatial distributions were greatly altered. The contents of moderately decomposable phosphorus, hardly decomposable phosphorus, and total phosphorus (TP) generally increased, while easily decomposable phosphorus content generally decreased. Compared with SA1, the contents of moderately decomposable phosphorus, hardly decomposable phosphorus and TP in SA2 increased by 11.5%, 9.7% and 10.5%, while those in SA3 increased by 24.8%, 13.2% and 13.5%, respectively. The distribution of phosphorus forms was greatly altered with increasing invasion years, which was dependent on the variations of key factors such as EC, pH value and grain composition. The implementation of regular mowing activities for S. alterniflora in the Minjiang River estuary in recent years, to some extent, reduced the return of phosphorus from residues to soils and decreased the availability of the easily decomposable phosphorus in soils.

[Effect of electroacupuncture on ocular surface sensory neuralgia and expression of P2X3 receptor and PKC in cornea and trigeminal ganglion of dry eye guinea pigs].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on ocular surface sensory neuralgia and the expression of P2X3 receptor (P2X3R) and protein kinase C(PKC)in cornea and trigeminal ganglion (TG) in dry eye disease (DED) guinea pigs, so as to explore its mechanism underlying improvement of ocular surface sensory neuralgia in DED. METHODS: Male British tricolor short haired guinea pigs were randomly divided into control, model, medication (pranoprofen), EA and sham acupuncture groups, with 8 guinea pigs in each group. The dry eye model was induced by subcutaneous injection of scopolamine hydrobromide solution (0.6 mg/0.2 mL,once daily) for 10 d. Guinea pigs in the medication group were treated by applying pranoprofen eye drops to eyes, 1 drop for one eye each time, three times a day. Guinea pigs of the EA group received EA stimulation (4 Hz/20 Hz,1 mA) of bilateral "Cuanzhu" (BL2) and "Taiyang" (HN5) and acupuncture at "Jingming" (BL1) "Sizhukong" (TE23), "Tongziliao" (GB1) for 15 min, once a day. Guinea pigs in the sham acupuncture group received blunt stimu-lation at the surface of the same acupoint with the tip of the acupuncture needle, once a day. All the treatments were conducted for 14 d. The corneal epithelium fluorescein staining score (0-3 points) was given according to the number of fluorescence-positive dots and flake-like coloration, the corneal mechanical perception thread (CMPT) detected using a corneal perception meter, and the palpebral fissure height measured. The number of sensory neurons in the cornea and TG was determined by using cholera toxin subunit B conjugated with Alexa Fluor 488 fluorescence labelling, and the expression levels of P2X3R and PKC in the cornea and TG detected by using immunohistochemistry and Western blot, separately. RESULTS: Compared with the control group, the corneal fluorescein staining score, immunoactivity and expression of P2X3R proteins in both cornea and TG, PKC proteins in TG were significantly increased (P<0.01), whereas the CMPT and the height of palpebral fissure and the number of TG neurons significantly decreased in the model group (P<0.05,P<0.01). In comparison with the model group, the fluorescein staining score in the medication and EA groups, the immunoactivity and expression of P2X3R in cornea and TG in the EA group, and that of TG PKC in the EA group and the sham acupuncture groups were significantly decreased (P<0.05, P<0.01), while the height of palpebral fissure and CMPT after EA and the number of labelling TG sensory neurons were remarkably increased in the EA group (P<0.01) rather than in the medication and sham acupuncture groups (P>0.05). CONCLUSION: EA can alleviate the damage of corneal epithelium and sensory neurons in dry eye model guinea pigs, which may be related to its functions in down-regu-lating the expression of P2X3R and PKC in the cornea and TG.

[Regulation Effect of Myeloid Leukemia No.1 Chinese Herb Medicine Prescription Combined with Chemotherapy on Th17 Cells in Bone Marrow of Patients with Acute Myeloid Leukemia].

OBJECTIVE: To explore the regulation effect of myeloid leukemia No.1 Chinese herb medicine prescription combined with chemotherapy on Th17 cells in bone marrow fluid of AML patients, so as to provide guidance for improving AML treatment effect and patients' long-term survival. METHODS: Seventy patients with AML who were hospitalized in Department of Hematology, Wuwei People's Hospital from April 2017 to August 2019 were selected and enrolled in AML group, 25 healthy volunteers were selected and enrolled in control group; then according to therapeutic regimen, AML patients were divided into 2 groups: combined therapy group (myeloid leukemia NO.1 Chinese herb medicine prescription combined with chemotherapy) and non-combined therapy group (chemotherapy alone). Flow cytometry was used to detect the ratio of CD3+ CD161+ IL-17+ IFN-γ+ T cells in bone marrow fluid, and ELISA was used to detect the vascular endothelial growth factor (VEGF) and interleukin-17 (IL-17) concentrations in bone marrow fluid. Statistical analysis was performed on the data with SPSS 22.0. RESULTS: The ratio of CD3+ CD161+ IL-17+ IFN-γ+ T cells, VEGF and IL-17 concentration in newly diagnosed and relapsed AML patients were significantly higher than those in the normal control group (P<0.001); while those in CR and DFS stage patients were significantly lower than those in newly diagnosed and relapsed patients (P<0.001), and the ratio of CD3+ CD161+ IL-17+ IFN-γ+ T cells, VEGF and IL-17 concentration in DFS patients with AML were not significantly different from those in the control group (P>0.05). The ratio of CD3+ CD161+ IL-17+ IFN-γ+ T cells, VEGF and IL-17 concentration in CR stage of AML patients treated with chemotherapy alone were significantly higher than those in the control group (P<0.05), but there was no difference between combined therapy group and the control group; the ratio of CD3+ CD161+ IL-17+ IFN-γ+ T cells, the concentration of VEGF and IL-17 in CR stage of AML patients treated with chemotherapy alone were higher than those of patients treated with combined therapy regimen (P<0.05). AML patients treated with combined therapy regimen had a significantly higher complete remission rate compared with patients received chemotherapy alone (P<0.05), but the recurrence rate was significantly lower (P<0.05). CONCLUSION: Th17 cells expression in bone marrow of newly diagnoses and relapsed AML patients significantly increase, and decrease significantly after treatment. Myeloid leukemia No.1 Chinese herb prescription combined with chemotherapy can significantly increase the CR rate and reduce the RL rate for AML.

PEGylated WS2 nanodrug system with erythrocyte membrane coating for chemo/photothermal therapy of cervical cancer.

The side effects of chemical drugs and multi-drug resistance are serious obstacles hindering efficient tumor therapy. Therefore, recently, the combination of chemo/photothermal therapy (CT/PT) has been adopted to address these issues using a low drug dosage. However, the development of multi-functional drug delivery systems with improved immune escape capability and enhanced drug accumulation at specific tumor tissues is still in its infancy. Herein, polyethylene glycol (PEG)-modified WS2 nanosheets (WS2-PEG) were used as a nanocarrier scaffold for doxorubicin (DOX, D) loading and near-infrared fluorescence probe indocyanine green (ICG, I) doping. After surface modification with the erythrocyte membrane (M) and targeted folic acid (FA) molecule, a new biomimetic system (WID@M-FA NPs) with high biocompatibility, prolonged cycle time (3.6-fold longer than WID NPs) and remarkable near-infrared photothermal function was developed for a targeted cervical cancer therapy. The in vitro assay indicated that the photothermal effects caused by ICG upon laser irradiation not only enhanced the cellular uptake of the drug, but also enhanced its tumor cell killing efficiency. Moreover, the targeted accumulation of DOX at the cervical cancer tissue and the synergistic chemo/photothermal therapy finally resulted in tumor elimination to more than 95% without side effects to the normal tissues in vivo. Thus, these excellent preclinical results indicate that WID@M-FA NPs may be an efficient therapeutic modality for the treatment of cervical cancer.

[Transcutaneous electrical acupoint stimulation for early enteral nutrition tolerance in patients with sepsis of gastrointestinal dysfunction: a multi-center randomized controlled trial].

OBJECTIVE: To compare the therapeutic effect of transcutaneous electrical acupoint stimulation (TEAS) combined with conventional western medicine and western medicine alone on early enteral nutrition tolerance in patients with sepsis. METHODS: Forty-nine patients with sepsis were randomly divided into an observation group (24 cases) and a control group (25 cases). The control group was treated with conventional western medicine, including limited fluid resuscitation, anti-infection and maintenance of hemodynamics, and enteral nutrition was provided when the hemodynamics became stable and the dosage of vasoactive drugs was reduced. The observation group, on the basis of the treatment of the control group, was treated with TEAS at Tianshu (ST 25), Shangjuxu (ST 37), Jiexi (ST 41), Diji (SP 8), Zusanli (ST 36), Zhongwan (CV 12) and Daheng (SP 15) when the enteral nutrition was provided (dilatational wave, 2 Hz/10 Hz, twice a day, 30 min each time), and the TEAS stopped when enteral nutrition was normal. The antral motility index (MI) was evaluated by ultrasonography on the first, third and fifth day into treatment in the two groups, and the duration from providing enteral nutrition to normal enteral nutrition was recorded. The acute gastrointestinal injury (AGI) classification, admission time of intensive care unit (ICU) and hospitalization time were compared. RESULTS: The antral MI on the third and fifth day into treatment in the observation group was significantly higher than that in the control group (P<0.05). The duration from providing enteral nutrition to normal enteral nutrition was (5.08±0.65) days in the observation group, which was shorter than (5.56±0.65) days in the control group (P<0.05). The improvement of AGI classification after treatment in the observation group was significantly superior to the control group (P<0.05). The hospitalization time in the observation group was shorter than that in the control group (P<0.05). CONCLUSION: TEAS combined with conventional western medicine treatment could significantly promote gastrointestinal peristalsis, improve the early intestinal nutrition tolerance in patients with sepsis, so as to shorten the time of reaching the standard of enteral nutrition and hospitalization time, which is better than conventional western medicine treatment alone.

[Effect of early acupoint electrical stimulation on lower limb muscle strength in patients with septic shock].

OBJECTIVE: To observe the effect of early acupoint electrical stimulation on the decline of lower limbs muscle strength in patients with intensive care unit-acquired weakness (ICU-AW) caused by septic shock. METHODS: A total of 58 patients with ICU-AW caused by septic shock were randomly divided into an observation group (28 cases, 1 case dropped off ) and a control group (30 cases, 2 cases dropped off ). Patients in both groups received routine basic treatment. In the observation group, acupoint electric stimulation therapy was added at Huantiao (GB 30), Futu (ST 32), Zusanli (ST 36), Xuanzhong (GB 39) and Taichong (LR 3). Unilateral point with electrodes were applied, the SDZ-Ⅱ electronic instrument (discontinuous wave, frequency in 2 Hz, strength in 5 mA) was connected and changed to the other side after 30 min of unilateral treatment.The treatment was given 2 times daily, continued for 7 d or until the medical research council (MRC) score being 54 points or more. The changes of lower limb muscle strength MRC score, modified Rankin scale (MRS) score, bilateral quadriceps thickness and gastrocnemius pinnate angle of both groups were observed before treatment and on discharge. The time of admission to ICU, time of hospitalization, mortality during hospitalization, and mortality 28 d after discharge were compared between the two groups. The MRS scores of the two groups were followed up 28 d after discharge. RESULTS: The MRC scores of lower limb muscle strength in the two groups on discharge were higher than those before treatment (P<0.05), and the MRS scores on discharge and 28 d after discharge in the two groups were lower than those before treatment (P<0.05). The MRC scores of lower limb muscle strength on discharge in the observation group were higher than thoes in the control group (P<0.05), and the MRS scores on discharge and 28 d after discharge in the observation group were lower than those in the control group (P<0.05). On discharge, bilateral quadriceps thickness and gastrocnemius pinnate angle in the two groups were increased compared with those before treatment (P<0.05), and thoese in the observation group was higher than the control group (P<0.05). There was no significant difference between the two groups in the time of admission to ICU, time of hospitalization, mortality during hospitalization, and mortality 28 d after discharge (P>0.05). CONCLUSION: Early acupoint electrical stimulation can improve the lower extremity muscle decline in patients with ICU-AW caused by septic shock.

[Effects of spatial expansion of Phragmites australis and Cyperus malaccensis on distributions and stocks of phosphorus in marshes in the Minjiang River estuary, China.] / 闽江河口芦苇与短叶茳芏空间扩展对湿地系统磷赋存特征的影响.

To investigate the distributions and stocks of total phosphorus (TP) in plant-soil systems of marshes in Shanyutan of the Minjiang River estuary and explore its influencing factors, Phragmites australis (PA) marsh and Cyperus malaccensis (CM) marsh before spatial expansion and ecotonal marsh (EM, P. australis and C. malaccensis in EM were denoted by PA' and CM', respectively) during spatial expansion were studied. Results showed that, as affected by spatial expansion, the contents of TP in both plant and soil in different marshes showed great variations. Compared with PA and CM marshes, soil TP in EM increased by 20.0% and 7.1%, respectively. The variation of soil TP in EM might be attributed to the alteration of soil particle composition, plant bio-mass and root/shoot ratio during spatial expansion. Except for leaves, TP in organs of P. australis in P. australis-C. malaccensis community (PA') was lower than that in C. malaccensis in P. australis-C. malaccensis community (CM'), due to the differences in absorption, utilization and translocation way of P among organs between the two species. The competition effect caused by spatial expansion greatly changed plant P allocation ratio of the two species. Compared with PA and CM, the allocations of P in roots and leaves of PA' increased, while only that in roots of CM' rose. During spatial expansion, the two species might adopt different adaptation strategies for P absorption and utilization to maintain their competitiveness. The PA might compete primarily by strengthening the P accumulation capacities of its roots and promoting leaf photosynthesis, whereas the CM might resist the spatial expansion of PA by increasing its underground biomass to enhance P absorption.

[Effect of ultrasound-guided acupoint electrical stimulation on diaphragmatic dysfunction associated with mechanical ventilation].

OBJECTIVE: To observe the effect of ultrasound-guided acupoint electrical stimulation on the patients with diaphragmatic dysfunction associated with mechanical ventilation in ICU. METHODS: Fifty-two patients were randomly divided into an observation group (26 cases, 3 cases dropping) and a control group (26 cases). Conventional treatment was given to all patients. On the basis of conventional treatment, acupoint electrical stimulation therapy was applied at Zhangmen (LR 3), Dabao (SP 21), Pishu (BL 20), Shenshu (BL 23), etc. In the observation group, the treatment was given for 30 min each time, 3 times a day for 7 days. Diaphragm thickening fraction (TFdi) was used as an index to guide the individualized setting of stimulation intensity and judge the effect, and the difference of mechanical ventilation time, ICU time, total hospitalization time, hospital mortality and reintubation rate between the two groups were observed. RESULTS: The mechanical ventilation time in the observation group was shorter than that in the control group (P<0.05). There was no significant difference in the ICU time and total hospitalization time between the two groups (P>0.05). During hospitalization, 2 patients died in the observation group and 3 patients died in the control group, there was no significant difference in hospital mortality (P>0.05). One patient in the observation group was reintubated and 8 patients in the control group (P<0.05). The use of acupoint electrical stimulation was a factor in shortening the mechanical ventilation time and reducing the reintubation events (P<0.05). CONCLUSION: Ultrasound-guided acupoint electrical stimulation can relieve ventilator-induced diaphragmatic dysfunction, reduce ventilator support time and reintubation events.

[Electroacupuncture improves insomnia by down-regulating peripheral benzodiazepine receptor expression in hippocampus, and up-regulating 5-HT, 5-HIAA, TNF-α and IL-1ß contents in hypothalamus in insomnia rats].

OBJECTIVE: To observe the therapeutic effect of electroacupuncture (EA) at five Back-Shu points on sleep, hippocampal peripheral benzodiazepine receptor (PBR) expression and hypothalamic 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1ß contents in insomnia rats, so as to explore its mechanisms underlying improvement of insomnia. METHODS: Forty male SD rats were randomly divided into control, model, EA and medication (Diazepam) groups (n=10 rats in each group). The insomnia model was established by intraperitoneal injection (i.p.) of para-Chlorophenylalanine (PCPA, 300 mg/kg) once daily for 2 days. EA (60 Hz, 1 mA) was applied to bilateral five Back-Shu points, i.e., Feishu (BL13), Xinshu (BL15), Ganshu (BL18), Pishu (BL20) and Shenshu (BL23) for 10 min, once daily for 6 days. Rats of the medication group were treated by gavage of Diazepam (0.92 mg/kg) once daily for 6 days. The sleep duration was recorded after i.p. of Pentobarbital Sodium (45 mg/kg). Histopathological changes of the hippocampus were displayed by H.E. staining. The contents of 5-HT, 5-HIAA, TNF-α and IL-1ß in the hypothalamus were assessed by using ELISA. The expression levels of PBR mRNA and protein in the hippocampus were detected by quantitative real-time PCR, immunohistochemistry and Western blot, separately. RESULTS: Following modeling, the sleep duration was considerably shortened in rats of the model group relevant to the control group (P<0.05). After the treatment, the sleep duration was significantly increased in both EA and medication groups relevant to the model group (P<0.05). After modeling, the hippocampal neurons were obviously decreased in number, and disordered in the arrangement, the levels of hypothalamic 5-HT, 5-HIAA, TNF-α, and IL-1ß were significantly down-regulated (P<0.05), and the expression of PBR mRNA and protein in hippocampus was obviously increased relevant to the control group (P<0.05). Following the treatment, the hippocampal neurons were increased in number and arranged regularly, and the contents of hypothalamic 5-HT, 5-HIAA, TNF-α, and IL-1ß were considerably increased (P<0.05), and the expression of PBR mRNA and protein in hippocampus was significantly decreased in both EA and medication groups relevant to the model group (P<0.05). The therapeutic effect of EA was comparable to that of medication in increasing sleep duration, body weight, 5-HT and 5-HIAA contents (P>0.05), and significantly superior to that of the medication in increasing TNF-α and IL-1ß levels (P<0.05), and considerably superior to that of medication in down-regulating PBR mRNA and protein expression (P<0.05). CONCLUSION: EA at five Back-Shu-points of the five Zang-organs can significantly improve the sleep in insomnia rats, which is closely associated with its effects in reducing the expression of PBR in hippocampus and up-regulating the levels of 5-HT, 5-HIAA, TNF-α and IL-1ß in hypothalamus.

Inhibitory effect of epicatechin gallate on protein glycation.

Protein glycation can produce the advanced glycation end products (AGEs), which has a potential health risk due to its relevance with diabetic complications. Searching for potent inhibitors to suppress hyperglycemia-induced protein glycation is of great significance. Here, we probed the anti-glycation potential of epicatechin gallate (ECG), an important ingredient of green tea, by spectroscopy and chromatography supplemented by molecular docking. It was found that ECG suppressed the carbonylation and the formation of amyloid cross-ß structures of bovine serum albumin (BSA) as well as the generation of AGEs in BSA-fructose model by 65.6%, 73.7% and 87.0%, resepctively. A combination of ECG with epigallocatechin gallate (EGCG) at molar ratios from 0.5:1 to 2:1 exhibited the interactive effects on AGEs formation from moderate antagonism to synergy. Moreover, ECG at 44.2 µg/mL showed a trapping efficiency of 81.6% for the important precursor methylglyoxal (MGO) of AGEs within 24 h, and the formed ECG-mono-MGO and ECG-di-MGO adducts were verified by LC-QqQ-MS analysis. The inhibitory effect of ECG on the protein glycation may be due to its MGO trapping and the specific binding with certain amino acid residues of BSA. These findings may provide the theoretical basis for the development of ECG as an inhibitor of protein glycation.

New insights into the binding mechanism between osthole and ß-lactoglobulin: Spectroscopic, chemometrics and docking studies.

Osthole, a type of coumarin derivative, owns many biological functions, but the poor water solubility and low bioavailability limit its usage in food and pharmaceutical fields. ß-Lactoglobulin (BLG) is a major protein in bovine milk whey which is usually used as a carrier of poorly soluble bioactive components. Researching the interaction between osthole and BLG can help us in understanding their binding mechanism and design an osthole delivery system using BLG as carrier protein to further promote the application of osthole in functional food and drug. Hence, this study was devoted to explore the binding properties of osthole with BLG and the effect on the protein structure by employing multispectroscopic approaches, chemometrics and molecular simulation studies. The results of the concentration profiles and pure spectra of the components resolved from highly overlapping spectral signals by the multivariate curve resolution-alternating least squares (MCR - ALS) algorithm verified the formation of an osthole-BLG complex. Moreover, osthole was found to strong quench the intrinsic fluorescence of BLG and the quenching mechanism was determined to be a static procedure. The binding constant of osthole with BLG at 298 K was (4.06 ±â€¯0.03) × 104 L mol-1. Hydrophobic forces and hydrogen bonds played major roles in the binding process. The analysis of synchronous fluorescence, circular dichroism and Fourier transform infrared spectra found that the presence of osthole caused BLG structure compact and the change in the polarity and hydrophobicity of the microenvironment around Tyr residues of the protein. Modeling docking predicted that osthole bound to the hydrophobic cavity of BLG through stable hydrogen bonds primarily with the amino acid residues Lys75 and Thr76.

Determination of the Five Main Terpenoids in Different Tissues of Wolfiporia cocos.

Wolfiporia cocos is a fungus containing triterpenoids and is widely used as an herbal medicine. However, it is unknown whether its main triterpenoid contents differ in different tissues. In this study, we identified dehydrotumulosic acid, polyporenic acid C, pachymic acid, dehydrotrametenolic acid, and dehydroeburicoic acid as the five main triterpenoids in W. cocos. We also systematically profiled the contents and distribution of these main triterpenoids in different tissues of W. cocos. High contents of all five triterpenoids were found in the surface layer of W. cocos. Intriguingly, we noted that the highest contents of the five triterpenoids were found in the surface layer of the sclerotium grown under pollution-controlled cultivation; the second-highest contents were found in the surface layer of the natural sclerotium. These results indicate that environmentally friendly cultivation of the sclerotium of W. cocos is a practical way to increase the productivity of W. cocos. In addition, our findings suggest that the triterpenoids may contribute to the pharmacological activity of W. cocos, and the surface layer of sclerotium in W. cocos might be a promising raw material for applications in health care and the development of functional medical products.

Investigation on risk factors of dry eye in type 2 diabetes patients / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To estimate the correlation between diabetic duration,blood glucose levels,plasma C-peptide and dry eye,and the risk factors for dry eye in patients with type 2 diabetes mellitus(T2 DM)METHODS:The clinical data of 51 patients (102 eyes) with type 2 diabetes diagnosed by the Department of Endocrinology,Jiangsu Provincial Hospital of Traditional Chinese Medicine was collected,in that 44 cases (88 eyes) of patients diagnosed with dry eye.Those patients were detected for the levels of glycosylated hemoglobin A1 c(HbA1 c),fasting blood-glucose (FBG),postprandial 2h blood-glucose (2h PBG),fasting plasma C-peptide and insulin,1h C-peptide and insulin.Corneal fluorescein staining(FL),tear break-up time(BUT) and Schirmer Ⅰ test (S Ⅰ t) were collected from all subjects.Compared biochemistry index and ocular surface index.The multiple Logistic regression was used to analyze the risk factors for dry eye in patients with r2DM.RESULTS:There was no significant differences between the patients with different diabetic duration,on BUT,S Ⅰ t,winking frequency,vision,FL and the scores of dry eye symptoms (P ＞ 0.05).HbA1c was significantly correlated with FL (P ＜ 0.05).There were significant differences in FL among patients with HbA1c in 8.1％ to 11.8％ (P＜0.01).FBG was significantly correlated with FL and winking frequency (P＜ 0.05).The 2h PBG was significantly correlated with tear secretion and vision (P＜0.05).Plasma C-peptide was significantly correlated with BUT (P＜0.05).There were significant differences in BUT among patients with 1h C-peptide in 3.3-5.5ng/mL(P＜0.05).FBG and plasma C-peptide in T2DM patients were risk factors for occurrence of dry eye(P＜0.05).CONCLUSION:Poor function of insulin secretion and poor control of blood glucose in T2DM patients are risk factors for dry eye.Both of them can decline tear film stability.High blood glucose levels easily lead to decrease of tear secretion,vision and corneal epithelial defect.

Repressing CD147 is a novel therapeutic strategy for malignant melanoma.

CD147/basigin, a transmembrane protein, is a member of the immunoglobulin super family. Accumulating evidence has revealed the role of CD147 in the development and progression of various cancers, including malignant melanoma (MM). MM is a malignancy of pigment-producing cells that causes the greatest number of skin cancer-related deaths worldwide. CD147 is overexpressed in MM and plays an important role in cell viability, apoptosis, proliferation, invasion, and metastasis, probably by mediating vascular endothelial growth factor (VEGF) production, glycolysis, and multi-drug resistance (MDR). As a matrix metalloproteinase (MMP) inducer, CD147 could also promote surrounding fibroblasts to secrete abundant MMPs to further stimulate tumor cell invasion. Targeting CD147 has been shown to suppress MM in vitro and in vivo, highlighting the therapeutic potential of CD147 silencing in MM treatment. In this review article, we discuss CD147 and its biological roles, regulatory mechanisms, and potential application as a molecular target for MM.

Effects of Optogenetic Activation of Corticothalamic Terminals in the Motor Thalamus of Awake Monkeys.

The role of the corticothalamic projection in the ventral motor thalamus remains poorly understood. Therefore, we studied the electrophysiological responses of neurons in the basal ganglia and cerebellar receiving-territories of the motor thalamus (BGMT and CbMT, respectively) using optogenetic activation of corticothalamic projections in awake rhesus macaques. After injections of viral vectors carrying the excitatory opsins ChR2 or C1V1 into the primary motor and premotor cortices of two monkeys, we used optrodes to light activate opsin-expressing neurons in cortex or their terminals in the thalamus while simultaneously recording the extracellular activity of neurons in the vicinity of the stimulation sites. As expected, light activation of opsins in the cerebral cortex evoked robust, short-latency increases in firing of cortical neurons. In contrast, light stimulation of corticothalamic terminals induced small-amplitude, long-latency increases and/or decreases of activity in thalamic neurons. In postmortem material, opsins were found to be expressed in cell bodies and dendrites of cortical neurons and along their corticothalamic projections. At the electron microscopic level, opsin labeling was confined to unmyelinated preterminal axons and small terminals that formed asymmetric synapses with dendrites of projection neurons or GABAergic interneurons in BGMT and CbMT and with neurons in the reticular thalamic nucleus. The morphological features of the transfected terminals, along with the long latency and complex physiological responses of thalamic neurons to their activation, suggest a modulatory role of corticothalamic afferents upon the primate ventral motor thalamus. SIGNIFICANCE STATEMENT: This study provides the first analysis of the physiological effects of cortical inputs on the activity of neurons in the primate ventral motor thalamus using light activation of opsin-containing corticothalamic terminals in awake monkeys. We found that selective light activation of corticothalamic terminals in contact with distal dendrites of thalamocortical neurons and GABAergic interneurons elicits complex patterns of slowly developing excitatory and inhibitory effects in thalamic neurons of the basal ganglia- and cerebellar-receiving regions of the motor thalamus. Our observations suggest a modulatory (instead of a "driver") role of the corticothalamic system in the primate ventral motor thalamus.