Biodegradable mesoporous nanocomposites with dual-targeting function for enhanced anti-tumor therapy.

Tumor-associated macrophages (TAMs), the main components of infiltrating leukocytes in tumors, often play a key role in promoting cancer development and progression. The tumor-specific microenvironment forces the phenotype of tumor-infiltrating to evolve in a direction favorable to tumor development, that is, the generation of M2-like TAMs. Consequently, the dual intervention of cancer cells and tumor microenvironment has become a research hotspot in the field of tumor immunotherapy. In this contribution, we developed pH-sensitive mesoporous calcium silicate nanocomposites (MCNs) encapsulated with indocyanine green (ICG) to enable the effective combination of photothermal therapy (PTT) and photodynamic therapy (PDT) triggered by the 808 nm near-infrared (NIR) light. The mannose and hyaluronic acid-grafted MCNs specifically targeted TAMs and tumor cells and promoted cell apoptosis both in vitro and in vivo. This paper revealed that irradiation of ICG loaded MCNs with NIR can produce a potent hyperthermia and induce abundant intracellular singlet oxygen generation in the target cells. These results suggest that the novel nanoplatform is believed to facilitate the delivery of chemotherapeutic agents to the tumor microenvironment (TME) to enhance the effects of tumor treatment.

Feishu Acupuncture Inhibits Acetylcholine Synthesis and Restores Muscarinic Acetylcholine Receptor M2 Expression in the Lung When Treating Allergic Asthma.

Acupuncture was proven beneficial in treating allergic inflammation. We aimed to explore the regulation underlying the effects of acupuncture on Feishu, an acupoint most commonly used in the acupuncture therapy for respiratory diseases, with respect to the system of sympathetic nerve neurotransmitter acetylcholine (Ach). Male Wistar rats were randomly grouping. No treatment was taken in the normal group. Allergic asthma was induced using ovalbumin on the model, Feishu acupuncture, and sham acupuncture groups; then control or acupuncture treatment lasting for 3 weeks was performed. Bronchoalveolar lavage fluid (BALF) from the four groups was examined. And pulmonary tissues were subjected to histological analysis with H&E staining; besides, immunofluorescent staining, quantitative PCR, and western blot were used to detect synthetase (ChAT) and Ach hydrolase (AchE), and its muscarinic receptors (mAchRs) M1-M3. There was inflammatory infiltration in the lung upon allergic asthma, which was alleviated by the Feishu acupuncture. The eosinophilic granulocytes, neutrophils, and lymphocytes in BALF from the Feishu acupuncture group were all significantly decreased compared with those of the model and sham acupuncture groups. The specific acupuncture on Feishu upon allergic asthma put down the pulmonary expression of ChAT, repaired at the level of gene expression the pulmonary expression of mAchR M1, and restored the pulmonary expression of mAchR M2 (especially in the bronchiolar epithelium) which has a role in inhibiting Ach release; while sham acupuncture had no effect. These results confirmed the therapeutic effects of Feishu acupuncture on allergic asthma, suggesting that the mechanisms may involve suppression of the Ach signal both from its synthesis and during its release.

Eplerenone restores 24-h blood pressure circadian rhythm and reduces advanced glycation end-products in rhesus macaques with spontaneous hypertensive metabolic syndrome.

Hypertension is often associated with metabolic syndrome (MetS), and serves as a risk factor of MetS and its complications. Blood pressure circadian rhythm in hypertensive patients has been suggested to contribute to cardiovascular consequences and organ damage of hypertension. But circadian changes of BP and their response to drugs have not been clearly investigated in non-human primates (NHPs) of MetS with hypertension. Here, we identified 16 elderly, hypertensive MetS rhesus monkeys from our in-house cohort. With implanted telemetry, we investigate BP changes and its circadian rhythm, together with the effect of antihypertensive drugs on BP and its diurnal fluctuation. MetS hypertensive monkeys displayed higher BP, obesity, glucose intolerance, and dyslipidemia. We also confirmed impaired 24-h BP circadian rhythm in MetS hypertensive monkeys. Importantly, Eplerenone, a mineralocorticoid receptor blocker, exerts multiple beneficial effects in MetS hypertensive monkeys, including BP reduction, 24-h BP circadian rhythm restoration, and decreased plasma concentration of inflammation factors and advanced glycation end-products. In summary, we identified a naturally-developed hypertensive MetS NHP model, which is of great value in the studies on pathogenesis of MetS-associated hypertension and development of novel therapeutic strategies. We also provided multiple novel mechanistic insights of the beneficial effect of Eplerenone on MetS with hypertension.

[Effects of electroacupuncture at "Neiguan" (PC 6) on sodium channel-related proteins in rats with ischemic myocardial injury].

OBJECTIVE: To explore the protective mechanism of electroacupuncture (EA) at "Neiguan" (PC 6) on ischemic myocardial injury, and to explain the response patterns and characteristics of the specific effect of acupoints along meridians in sodium channel in the level of cardiac organ. METHODS: A total of 60 SPF male rats were randomly divided into a blank group, a model group, a non-acupoint group, a Neiguan group and a Lieque group, 12 cases in each one. Except the blank group, rats in the remaining group were treated with subcutaneous injection of isoprenaline to establish the model of myocardial ischemia. Rats in the Neiguan group, Lieque group and non- acupoint group were treated with EA, dilatational wave, with a frequency of 2 Hz/20 Hz. The intensity was 2-3 mA. The needles were retained for 20 min per time, once a day for consecutive 7 days. In the blank group and control group, the rats were grasped and fixed at the treating time each day. The western-blot method was used to test the expression of voltage-gated sodium channel alpha subunit (Nav 1.5), protein tyrosine kinase (PTKs) and protein tyrosine phosphatase (PTPs). RESULTS: The expression of Nav 1.5 and PTKs in the model group was lower than that in the blank group (both P<0. 01); the expression in the Neiguan group and Lieque group was higher than that in the model group (all P < 0.01); the expression of Nav 1.5 and PTKs in the Neiguan group was higher than that in the Lieque group (both P < 0.01). The expression of PTPs in the model group and non-acupoint group was higher than that in the blank group (both P < 0.01); the expression of PTPs in the Neiguan group and Lieque group was significantly down-regulated, which was lower than the model group (both P < 0.01); the down-regulation in the Neiguan group was significantly different from that in the Lieque group (P < 0.05). CONCLUSION: EA at "Neiguan" (PC 6), by down-regulating the expression of PTPs, up-regulating the expression of Nav 1.5 and PTKs, is likely to achieve the aim of regulation on sodium channel activity and calcium overload, further to improve myocardial ischemia, which provides experimental basis for the theory of the specific effect of acupoints along meridians.

[Methods for Accurately Locating "Feishu" (BL 13) in Rats].

"Feishu" (BL 13) is a commonly used acupoint in experimental study,'but its definite position has not been described in current books about acupoints. In the'present paper, the authors introduced three methods for determining the anatomical location of "Feishu" (BL 13) acupoint in rats. The Wistar rats (200 ± 20) g were anesthetized with 10% chloral hydrate first, and then, fixed at the prone position. The highest point of the spinous process of the second thoracic vertebra was used as the marker for positioning the spinal vertebra. The point, 7 mm lateral to the site below the spinous process of the 3rd thoracic vertebra is just the "Feishu" (BL 13). In order to determine the 3rd thoracic vertebra, three methods are recommended to be adopted: 1) finding the 5th thoracic vertebra process parallel to the inferior angle of the scapula first, then going upwards to find the 3rd thoracic vertebra , 2) finding the 1st thoracic vertebra along the 1st rib, then going downward to find the 3rd thoracic vertebra, and 3) taking the 6th lumbar vertebra (parallel to the hip-joint) as the bone-marker and going upward to find the 3rd thoracic vertebra. When "Feishu" (BL 13) punctured, a filiform needle was straight inserted into the skin, then downward to the subcutaneous tissue, muscular layer (including the trunk cutaneous muscle, trapezius muscle, broadest muscle of back, serratus muscle of back, atalanto-longest muscle, longest muscle of neck, longest muscle of thorax, lumbar longest muscle, the medial part of the longest muscle, cervico-spinal muscle of back-, semispinal muscle of neck, biventer cervicis, and multifidus muscles) about 6 mm in depth.

Effects of Deep Electroacupuncture Stimulation at "Huantiao" (GB 30) on Expression of Apoptosis-Related Factors in Rats with Acute Sciatic Nerve Injury.

SD rats were randomly divided into normal control, model, deep EA, and shallow EA groups. The model was established by mechanical clamping of the sciatic nerve stem. For deep and shallow EA, the needles were inserted into "Huantiao" (GB 30) by about 16 mm and 7 mm, respectively, once daily for 14 days. The results showed that, compared with the normal control group, the nerve-muscle excitability of rat's hip muscle decreased and the nerve conduction velocity of sciatic nerve slowed down in the model group; meanwhile, the number of apoptotic cells and the expression level of Bax protein in the injured nerve increased significantly, and the expression level of Bcl-2 protein and the ratio of Bcl-2/Bax decreased considerably. Compared with the model group, the indices mentioned above were reversed in the two treatment groups, and the changes in the deep EA group were more significant than those in the shallow EA group. These results indicate that EA stimulation at GB 30 can improve the function of injured sciatic nerve, which is closely associated with its effects in upregulating the expression of apoptosis inhibitive factor Bcl-2 and downregulating apoptosis promotive factor Bax. Deep EA is relatively better.

[Effect of electroacupuncture stimulation of "Neiguan" (PC 6) on expression of alpha- and beta-subunit proteins of voltage-gated sodium channels in rats with myocardial ischemia].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation of "Neiguan" (PC 6) on expression of sodium (Nav) channel α-subunit 1.5 and Nav ß-subunits ß 1-ß 4 (the known myocardial sodium channel proteins) in acute myocardial ischemia (AMI) rats so as to explore its mechanisms underlying protection of ischemic myocardium. METHODS: Sixty SD male rats were randomly divided into normal control, AMI model, Neiguan (PC 6), Lieque (LU 7) and non-acupoint groups. The AMI model was established by intravenous injection of Isoprenaline (85 mg/kg) once daily for 2 days. Myocardial Nav 1.5 and Nav ß 1, ß 2, ß 3, ß 4 protein expression levels were detected by Western blot. RESULTS: In comparison with the control group, myocardial Nav 1.5, ß 1, ß 2, ß 3 and ß 4 protein expression levels were significantly down-regulated in the AMI model group (P<0.01). After EA stimulation, compared with the model group, the expression levels of Nav 1.5, Nav ß 1, ß 2, ß 3 and ß 4 protein expression levels were significantly up-regulated in the Neiguang (PC 6) and Lieque (LU 7) groups (P<0.01) rather than in the non-acupoint group (P>0.05). There was no statistical significance between the Neiguan (PC 6) and Lieque (LU 7) groups in ß 4 protein expression level (P>0.05). CONCLUSION: EA stimulation of both PC 6 and LU 7 can significantly reverse AMI-induced down-regulation of myocardial Nav 1.5, ß 1, ß 2, ß 3 and ß 4 protein expression levels in AMI rats, which might contribute to its function in improving AMI by reducing calcium overload.

[Effect of deep electroacupuncture stimulation of "Huantiao" (GB 30) on changes of function and nerve growth factor expression of the injured sciatic nerve in rats].

UNLABELLED: OBJECTIVE; To observe the effect of deep electroacupuncture (EA) stimulation of "Huantiao"(GB 30) on the functional and pathological changes and nerve growth factor (NGF) expression of the damaged sciatic nerve in rats, so as to study its mechanisms underlying reliving sciatica. METHODS: Forty-eight SD rats were randomly divided into normal, model, deep EA and shallow EA groups (n = 12 in each group). The sciatic nerve injury model was established by mechanical clamp of the sciatic nerve stem. For deep and shallow EA, the acupuncture needles were inserted into GB 30 about 16 mm and 7 mm, respectively. The EA treatment was given 20 min, once daily for 14 days. The evoked potentials of the injured sciatic nerve stem responding to electrical stimulation were recorded by using a biophysiological experimental system for calculating the motor conduction velocity. Pathological changes of the sciatic nerve were displayed by H. E. stain. The expression of NGF and Fos proteins was detected by immunohistochemistry. RESULTS: In comparison with the normal group, the conduction velocity and the amplitude of the evoked potentials of the sciatic nerve were significantly decreased in the model group (P < 0.05). Following EA, both conduction velocity and amplitude of the evoked potentials in the deep EA group, and the conduction velocity in the shallow EA group were considerably increased (P < 0.05). The therapeutic effects were significantly better in the deep EA group than that in the shallow EA group (P < 0.05). No significant differences were found between the shallow EA and model groups in the amplitude of evoked potentials (P > 0.05), and no significant changes of latencies of the evoked potentials inthe four groups (P > 0.05). In the model group, the disorganized nerve fibers axons, myelin and Schwann cells of the damaged sciatic nerve were found, which became milder in the EA groups particularly in the deep EA group. In regard to the NGF and Fos immunoactivity of the injured sciatic nerve, the expression levels of both NGF and Fos proteins were obviously higher in the model group than in the normal group (P < 0.05). After EA stimulation, NGF expression was further significantly up-regulated in both deep and shallow EA groups (P < 0.05), and Fos protein expression was notably down-regulated in the deep and shallow EA groups (P < 0.05). The expression of NGF was significantly higher and Fos protein was obviously lower in the deep EA group than in the shallow EA group (P < 0.05). CONCLUSION: Deep EA stimulation of GB 30 can improve the pathological changes and function of the injured sciatic nerve in the rat, which is closely associated with its effects in up-regulating NGF expression and down-regulating Fos expression. The deep EA is relatively better in the therapeutic effect. These facts may be one of the mechanisms of EA in relieving sciatica in clinic practice.

Liver-directed neonatal gene therapy prevents cardiac, bone, ear, and eye disease in mucopolysaccharidosis I mice.

Mucopolysaccharidosis I (MPS I) due to deficient alpha-L-iduronidase (IDUA) activity results in accumulation of glycosaminoglycans in many cells. Gene therapy could program liver to secrete enzyme with mannose 6-phosphate (M6P), and enzyme in blood could be taken up by other cells via the M6P receptor. Newborn MPS I mice were injected with 10(9) (high dose) or 10(8) (low dose) transducing units/kg of a retroviral vector (RV) expressing canine IDUA. Most animals achieved stable expression of IDUA in serum at 1240 +/- 147 and 110 +/- 31 units/ml, respectively. At 8 months, untreated MPS I mice had aortic insufficiency, increased bone mineral density (BMD), and reduced responses to sound and light. In contrast, MPS I mice that received high-dose RV had normal echocardiograms, BMD, auditory-evoked brain-stem responses, and electroretinograms. This is the first report of complete correction of these clinical manifestations in any model of mucopolysaccharidosis. Biochemical and pathologic evaluation confirmed that storage was reduced in these organs. Mice that received low-dose RV and achieved 30 units/ml of serum IDUA activity had no or only partial improvement. We conclude that high-dose neonatal gene therapy with an RV reduces some major clinical manifestations of MPS I in mice, but low dose is less effective.