Cytotype distribution and chloroplast phylogeography of the Actinidia chinensis complex.

BACKGROUND: Plant phylogeographic studies of species in subtropical China have mainly focused on rare and endangered species, whereas few studies have been conducted on taxa with relatively wide distribution, especially polyploid species. We investigated the cytotype and haplotype distribution pattern of the Actinidia chinensis complex, a widespread geographically woody liana with variable ploidy in subtropical China comprising two varieties, with three chloroplast fragments DNA (ndhF-rpl132, rps16-trnQ and trnE-trnT). Macroevolutionary, microevolutionary and niche modeling tools were also combined to disentangle the origin and the demographic history of the species or cytotypes. RESULTS: The ploidy levels of 3338 individuals from 128 populations sampled throughout the species distribution range were estimated with flow cytometry. The widespread cytotypes were diploids followed by tetraploids and hexaploids, whereas triploids and octoploids occurred in a few populations. Thirty-one chloroplast haplotypes were detected. The genetic diversity and genetic structure were found to be high between varieties (or ploidy races) chinensis and deliciosa. Our results revealed that these two varieties inhabit significantly different climatic niche spaces. Ecological niche models (ENMs) indicate that all varieties' ranges contracted during the Last Inter Glacial (LIG), and expanded eastward or northward during the Last Glacial Maximum (LGM). CONCLUSIONS: Pliocene and Plio-Pleistocene climatic fluctuations and vicariance appear to have played key roles in shaping current population structure and historical demography in the A. chinensis complex. The polyploidization process also appears to have played an important role in the historical demography of the complex through improving their adaptability to environmental changes.

Bakuchiol exhibits anti-metastasis activity through NF-κB cross-talk signaling with AR and ERß in androgen-independent prostate cancer cells PC-3.

Androgen-independent prostate cancer (PCa) is a developed tumor derived from the local androgen dependent PCa, which often affects elderly men. Psoralea corylifolia L, a traditional Chinese medicine, has been widely used for PCa treatment as an important part of a common prescription, while the mechanism remains unclear. Our study was aimed to investigate the tumor-inhibitory effect of its main component bakuchiol in androgen-independent PCa cell line PC-3 cells. Bakuchiol significantly suppressed PC-3 cell proliferation and migration; the expressions of PCNA and MMP-9 were consistently down regulated as well. Meanwhile, both the constitutive and LPS-induced NF-κB activation were significantly inhibited by bakuchiol. The inhibitory effects of bakuchiol on cell proliferation, migration and invasion were recovered when LPS were added together with bakuchiol. SiRNA against androgen receptor (AR) or estrogen receptor ß (ERß) were transfected and the regulation of bakuchiol-suppressed proliferation, invasion, NF-κB signaling and MMP-9 secretion in response to LPS were blocked. Taken together, our data demonstrated that bakuchiol inactivated NF-κB signaling via AR and ERß, which contributes to inhibition of PC-3 cell proliferation and migration, indicating that bakuchiol is one of the key component from P. corylifolia L for PCa treatment and has a potential as anti-prostate cancer drug candidates.

Network pharmacology exploration reveals endothelial inflammation as a common mechanism for stroke and coronary artery disease treatment of Danhong injection.

Although Danhong injection (DHI) is the most widely prescribed Chinese medicine for both stroke and coronary artery disease (CAD), its underlying common molecular mechanisms remain unclear. An integrated network pharmacology and experimental verification approach was used to decipher common pharmacological mechanisms of DHI on stroke and CAD treatment. A compound-target-disease & function-pathway network was constructed and analyzed, indicating that 37 ingredients derived from DH (Salvia miltiorrhiza Bge., Flos Carthami tinctorii and DHI) modulated 68 common targets shared by stroke and CAD. In-depth network analysis results of the top diseases, functions, pathways and upstream regulators implied that a common underlying mechanism linking DHI's role in stroke and CAD treatment was inflammatory response in the process of atherosclerosis. Experimentally, DHI exerted comprehensive anti-inflammatory effects on LPS, ox-LDL or cholesterol crystal-induced NF-κB, c-jun and p38 activation, as well as IL-1ß, TNF-α, and IL-10 secretion in vascular endothelial cells. Ten of 14 predicted ingredients were verified to have significant anti-inflammatory activities on LPS-induced endothelial inflammation. DHI exerts pharmacological efficacies on both stroke and CAD through multi-ingredient, multi-target, multi-function and multi-pathway mode. Anti-endothelial inflammation therapy serves as a common underlying mechanism. This study provides a new understanding of DHI in clinical application on cardiovascular and cerebrovascular diseases.

Decreased plasma levels of lipoxin A4 in children with autism spectrum disorders.

The aim of this study was to evaluate the plasma levels of lipoxin A4 (LXA4), a mediator involved in the resolution of inflammation in Chinese children with autism spectrum disorders (ASD). From January 2013 to June 2014, a total of 150 children (75 confirmed ASD cases and 75 their age-matched and sex-matched control cases) participated in this study after consent was obtained from their parents. Clinical information was collected. Plasma levels of LXA4 were measured at baseline. The severity of ASD was assessed at admission using the Childhood Autism Rating Scale total score. The results indicated that the mean plasma levels of LXA4 were significantly lower in autistic children compared with the normal children (P<0.0001). There was a significant negative relationship between circulating LXA4 levels and severity of autism evaluated by Childhood Autism Rating Scale scores (P=0.006) after adjustment for the possible covariates. On the basis of the receiver operating characteristic curve, the optimal cutoff value of plasma LXA4 levels as an indicator for an auxiliary diagnosis of ASD was projected to be 81.5 pg/ml, which yielded a sensitivity of 90.7% and a specificity of 76.0%, with the area under the curve at 0.911 (95% confidence interval, 0.867-0.955). These results suggested that autistic children had lower plasma LXA4 levels, suggesting an increased susceptibility to recurring inflammation in these samples.

Chrysophanol attenuates lead exposure-induced injury to hippocampal neurons in neonatal mice.

Previous studies have shown that chrysophanol protects against learning and memory impairments in lead-exposed adult mice. In the present study, we investigated whether chrysophanol can alleviate learning and memory dysfunction and hippocampal neuronal injury in lead-exposed neonatal mice. At the end of lactation, chrysophanol (0.1, 1.0, 10.0 mg/kg) was administered to the neonatal mice by intraperitoneal injection for 15 days. Chrysophanol significantly alleviated injury to hippocampal neurons and improved learning and memory abilities in the lead-poisoned neonatal mice. Chrysophanol also significantly decreased lead content in blood, brain, heart, spleen, liver and kidney in the lead-exposed neonatal mice. The levels of malondialdehyde in the brain, liver and kidney were significantly reduced, and superoxide dismutase and glutathione peroxidase activities were significantly increased after chrysophanol treatment. Collectively, these findings indicate that chrysophanol can significantly reduce damage to hippocampal neurons in lead-exposed neonatal mice.

[Preparation process of rutacarpine-hydroxypropyl-beta-cyclodextrin inclusion complex].

Rutaecarpine (Rut) is a type of indole quinazoline alkaloid exracted from Ruticarpum. Studies showed that Rut has a wide range of pharmacological effects, such as anti-hypertension, anticancer, anti-inflammation, anti-thrombus formation. Currently, many scholars are committed to developing it into a new antihypertensive and anti-inflammatory drug with all new mechanisms. But studies found that Rut is a highly fat-soluble drug with low water and oil solubility. Its high insolubility is the main obstacle in its oral absorption and application, which greatly reduced its bioavailability. Therefore, hydroxypropyl-beta-cyclodextrin (HP-beta-CD) was used as the inclusion material to prepare Rut-HP-beta-CD inclusion complex in this experiment, in order to increase its water solubility and bioavailability. In this experiment, the inclusion complex was prepared by the stirring-freeze-dry method. The preparation process was optimized by the orthogonal test, with the inclusion rate as the index, and molar ratio between host and guest molecules, inclusion temperature, time and stirring speed as the impacting factors. Moreover, the inclusion complex was verified by detecting the apparent solubility, thin layer chromatography, microscopic identification, melting point detection and dissolution study. The results showed that under the conditions of the molar ratio between Rut and HP-beta-CD of 1: 1, temperature at 60 degrees C, inclusion time of 4h and stirring speed at 600 r x min(-1), the inclusion rate of Rut-HP-beta-CD reached 91.04%. Therefore, the preparation process of Rut-HP-beta-CD inclusion under the optimum conditions is simple and feasible, with a highest inclusion rate and reproducibility, and could significantly improve Rut's solubility and bioavailability, and provide a reliable experimental basis for its clinical application.

Ultrasound-assisted extraction of total phenolic compounds from Inula helenium.

Ultrasound-assisted extraction (UAE) of phenolic compounds from Inula helenium was studied. Effects of ethanol concentration, ultrasonic time, solid-liquid ratio, and number of extractions were investigated. An orthogonal array was constructed to optimize UAE process. The optimized extraction conditions were as follows: ethanol concentration, 30%; solid-liquid ratio, 1 : 20; number of extractions, 2 times; extraction time, 30 min. Under the optimal conditions, the yield of total phenolic compounds and chlorogenic acid was 6.13 ± 0.58 and 1.32 ± 0.17 mg/g, respectively. The results showed that high amounts of phenolic compounds can be extracted from I. helenium by ultrasound-assisted extraction technology.

Neuroprotective effects of rutaecarpine on cerebral ischemia reperfusion injury.

Rutaecarpine, an active component of the traditional Chinese medicine Tetradium ruticarpum, has been shown to improve myocardial ischemia reperfusion injury. Because both cardiovascular and cerebrovascular diseases are forms of ischemic vascular disease, they are closely related. We hypothesized that rutaecarpine also has neuroprotective effects on cerebral ischemia reperfusion injury. A cerebral ischemia reperfusion model was established after 84, 252 and 504 µg/kg carpine were given to mice via intraperitoneal injection, daily for 7 days. Results of the step through test, 2,3,5-triphenyl tetrazolium chloride dyeing and oxidative stress indicators showed that rutaecarpine could improve learning and memory ability, neurological symptoms and reduce infarction volume and cerebral water content in mice with cerebral ischemia reperfusion injury. Rutaecarpine could significantly decrease the malondialdehyde content and increase the activities of superoxide dismutase and glutathione peroxidase in mouse brain. Therefore, rutaecarpine could improve neurological function following injury induced by cerebral ischemia reperfusion, and the mechanism of this improvement may be associated with oxidative stress. These results verify that rutaecarpine has neuroprotective effects on cerebral ischemia reperfusion in mice.

Single administration of lesion-limited high-dose (TURBO) ultraviolet B using the excimer laser: clinical clearing in association with apoptosis of epidermal and dermal T cell subsets in psoriasis.

BACKGROUND/PURPOSE: Development of effective therapy for psoriasis is confounded by numerous factors contributing to disease pathogenesis, including pathogenic immunocytes which appear to drive epidermal keratinocyte hyperproliferation. Our objective was to study clinical and biomarker effects of a single dose of TURBO laser UVB (308 nm) applied directly to psoriatic plaques. METHODS: Eighteen patients with chronic plaque psoriasis received a single dose of 10 minimal erythema dose (MED) UVB and were followed for 8 weeks. Keratome and punch biopsies were assessed for T cell depletion and apoptosis following a single 308-nm dose of UVB. RESULTS: Patients demonstrated clinical improvement as indicated by decreased global Psoriasis Area and Severity Index scores and reduced numbers of pathogenic memory/effector T cells infiltrating lesional epidermis and dermis. Consistent with apoptosis induction, caspase activation increased in lesional T cells after treatment. CONCLUSION: We conclude that a single 10 MED dose of TURBO UVB is effective at reducing the severity and extent of psoriatic lesions. We hypothesize that the reason a single treatment is sufficient to clear a psoriatic plaque is that the 10 MED dose is able to deliver sufficient photons to a microanatomic area of the lesion where susceptible pathogenic T cell mechanisms are operative.

[Inhibition of elicitation of allergic contact dermatitis by topical use of Z-VAD-FMK, a broad caspase inhibitor: experiment in mice].

OBJECTIVE: To explore the effects of N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-FMK), a broad caspase inhibitor, on the elicitation of murine allergic contact dermatitis (ACD) and examine the effects on T lymphocytes. METHODS: 1-fluoro-2,4-dinitrobenzene (DNFB) was used to establish the classical murine model of ACD. Different concentrations of Z-VAD-FMK were applied before ear provocation. Several parameters were detected, including ear swelling degree, weight differences and thickness of ear tissue under microscope between 2 ears. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of Th1 cytokines (INF-γ and IL-2) in ear tissues. Real-time reverse transcription-polymerase chain reaction (RT-PCR) was used to detect their levels of mRNA and the results were shown as "copies relative to one million housekeeping genes". Local lymph node assay (LLNA) was conducted. Bromodeoxyuridine-flow cytometry was used to detect the proliferation of T lymphocytes in local lymph node and flow cytometry to detect the activation of T lymphocytes. RESULTS: The right ear swelling degree, weight differences and thickness between two ears in the 1.25 mmol/L Z-VAD-FMK group were (12.6 ± 1.2)×10(-2) mm, (3.1 ± 0.2) mg, and (12.1 ± 1.1)×10(-2) mm respectively. And they were all significantly lower than those of the negative control group((17.4 ± 1.6)×10(-2) mm, (4.2 ± 0.3) mg, (16.7 ± 1.5)×10(-2) mm;q = 3.25, 2.98, 3.12, all P < 0.05). The levels of INF-γ and IL-2 in the ear skin lesions of 1.25 mmol/L Z-VAD-FMK group were (856 ± 45) and (167 ± 12) pg/ml respectively and they were both significantly lower than those of the negative control group ((1180 ± 58) and (225 ± 16) pg/ml; q = 3.11, 3.14, both P < 0.05). The mRNA levels of the above two cytokines were 152 ± 12 and 96 ± 8 respectively and they were both significantly lower than those of the negative control group (220 ± 15 and 156 ± 11;q = 3.15, 3.42, both P < 0.05). In LLNA, the mean intensity of BrdU in T lymphocytes of 1.25 mmol/L Z-VAD-FMK-treated group was significantly weaker than that of the negative control group (185 ± 15 vs 298 ± 21, q = 3.02, P < 0.05). The percent of activation markers-positive T lymphocytes of the Z-VAD-FMK group were 7.8% ± 0.7%, 9.8% ± 0.8% and 31.2% ± 2.8% respectively and they were all significantly lower than those of the negative control group (10.5% ± 1.0%, 14.5% ± 1.1%, 46.5% ± 3.2%, q = 3.16, 3.52, 3.11, all P < 0.05). CONCLUSION: Topical use of Z-VAD-FMK prior to ear provocation can suppress the proliferation and activation of T lymphocytes in both skin tissues and local lymph nodes and thus result in the inhibitory effect of allergic contact dermatitis.

Total glucosides of Paeonia lactiflora Pall inhibit vascular endothelial growth factor-induced angiogenesis.

OBJECTIVE: To evaluate the anti-angiogenesis effect of total glucosides of Paeonia lactiflora Pall. METHODS: In this study, we determined the effect of TGP on the proliferation of human vascular endothelial cells through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and fluorescence-activated cell sorting analysis. A migration assay and a tube formation assay were used to investigate the migration properties and tube formation abilities of human vascular endothelial cells after being treated with TGP. Furthermore, the in vivo anti-angiogenic ability of TGP was determined through a chick chorioallantoic membrane assay. RESULTS: TGP (12.5, 62.5, and 312.5 microg/ml) resulted in a dose-dependent reduction in the proliferation of endothelial cells. This inhibition effect began 6h after treatment and lasted at least 24h. Fluorescence-activated cell sorting analysis data showed an accumulation of cells in the G0/G1 phase of the cell cycle, which exhibited apoptotic features indicative of cell death. The migration properties and tube forming abilities of endothelial cells were dramatically inhibited by the TGP extract. CONCLUSION: Our results show that TGP can inhibit angiogenesis in vitro and in vivo.

Photodynamic therapy with the silicon phthalocyanine pc 4 induces apoptosis in mycosis fungoides and sezary syndrome.

Our current focus on the effects of Photodynamic Therapy (PDT) using silicon phthalocyanine Pc 4 photosensitizer on malignant T lymphocytes arose due to preclinical observations that Jurkat cells, common surrogate for human T cell lymphoma, were more sensitive to Pc 4-PDT-induced killing than epidermoid carcinoma A431 cells. Mycosis fungoides (MF) as well as Sezary syndrome (SS) are variants of cutaneous T-cell lymphoma (CTCL) in which malignant T-cells invade the epidermis. In this study, we investigated the cytotoxicity of Pc 4-PDT in peripheral blood cells obtained from patients with SS and in skin biopsies of patients with MF. Our data suggest that Pc 4-PDT preferentially induces apoptosis of CD4(+)CD7(-) malignant T-lymphocytes in the blood relative to CD11b(+) monocytes and nonmalignant T-cells. In vivo Pc 4-PDT of MF skin also photodamages the antiapoptotic protein Bcl-2.

A 2940 nm fractional photothermolysis laser in the treatment of acne scarring: a pilot study in China.

OBJECTIVE: To evaluate the efficacy and safety of a 2940 nm fractional photothermolysis laser in the treatment of acne scarring in Chinese people. METHODS: Twenty-six patients with moderate-to-severe atrophic scarring were treated with a 2940 nm-wavelength fractional photothermolysis laser. RESULTS: All patients had encouraging results. Both skin elasticity and moisture content increased significantly after five treatments. In post-treatment evaluations, both the patients treated, as well as an independent group of physicians each scored the atrophic scar improvement as significant. CONCLUSION: The 2940 nm fractional photothermolysis laser is safe and effective in the treatment of acne scarring.

Fractional 1320 nm Nd : YAG laser in the treatment of acne vulgaris: a pilot study.

Thirty-five patients with moderate to severe acne were treated with a fractional 1320 nm neodymium : yttrium aluminum garnet (Nd : YAG) laser. These patients received six treatment sessions at a 2-week interval. Inflammatory and non-inflammatory lesions were counted before and after treatment. Fractional 1320 nm Nd : YAG laser therapy was well tolerated, resulting in the reduction of inflammatory lesions by 57% (P<0.05) and the reduction of non-inflammatory lesions by 35% (P<0.05). A significant reduction in the skin sebum level by 30% (P<0.05) was also noted after treatment.

[Attenuation of allergic contact dermatitis by Z-VAD-FMK, a broad caspase inhibitor: experiment with mice].

OBJECTIVE: To investigate the effects of N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-FMK), a broad caspase inhibitor, on allergic contact dermatitis (ACD). METHODS: A Balb/c mouse was killed and its back skin was peeled, put into a Franz diffusion cell, and smeared with Z-VAD-FMK. High performance liquid chromatography was used to examine the permeation rate. Dinitro-fluorobenzene (DNFB) was applied on the depilated back of mice as inducer and then on the back-side of right ear as provocative agent 6 days later so as to establish mouse ACD models. Two hours after the provocation 24 mouse models were divided into 4 equal groups to undergo the application of Z-VAD-FMK at the concentration of 0.1, 0.5, 2.5, and 5 mmol/L on the back-side of right ear twice. PBS was applied in the negative control group. Then the mice were killed with their 2 ears cut off. Microscopy was used to measure the right ear swelling degree, and differences in weight and thickness between the 2 ears. ELISA was used to detect the levels of IL-2 and INF-gamma in the ear tissues. RESULTS: The transdermal permeation rates of Z-VAD-FMK at the time points 6, 12, and 24 h were 2.15%, 9.62%, and 12.85% respectively. The right ear swelling degree, and differences in weight and thickness between the 2 ears in the 2.5 mmol/L Z-VAD-FMK group were (12.5 +/- 1.4) x 10(-2) mm, (3.2 +/- 0.3) mg, and (11.8 +/- 1.3) x 10(-2) mm respectively, all significantly lower than those of the negative control group [(19.1 +/- 2.0) x 10(-2) mm, (4.3 +/- 0.4) mg, and (16.8 +/- 1.7) x 10(-2) mm, all P < 0.05]. The IL-2 and INF-gamma levels in the ear skin lesion of the 2.5 mmol/L Z-VAD-FMK group were (148 +/- 10) and (650 +/- 45) pg/ml respectively, both significantly lower than those of the negative control group [(205 +/- 18) and (1030 +/- 58) pg/ml, both P < 0.05]. CONCLUSION: Z-VAD-FMK can permeate through mouse skin and inhibit the activation and proliferation of T lymphocytes, leading to the inhibitory effect of contact allergic reaction.

Cutaneous lesions and visceral involvement of tuberous sclerosis.

BACKGROUND: Tuberous sclerosis (TS) is an autosomal dominant disorder with a significant range of clinical expressions. The involvement of vital organs, such as the brain, kidney, heart and lung is the main cause of death in patients with TS. The aim of this study is to summarize the characteristic cutaneous features and common extracutaneous involvement of TS, which are helpful to the early detection of visceral involvement. METHODS: The analyzed clinical data from 78 patients with TS included those from detailed history, physical and dermatological examination, cranial computed tomography (CT) and magnetic resonance imaging (MRI), abdominal ultrasonography, chest roentgenography, hand and foot X-ray and ophthalmologic examination. RESULTS: The skin, brain and kidney were involved frequently in TS patients. Hypomelanotic macules were the most common and earliest cutaneous lesions. Their number was more than 3 in 81.5% of the patients. They were followed by facial angiofibromas and Shangreen's patch in a decreasing frequency. Forehead plaque, facial angiofibromas and Shagreen's patch appeared in patients at mean age of 2.6, 6.0 and 8.1 years respectively. Cranial CT showed a high positive rate in TS patients. CONCLUSIONS: Cutaneous features of TS are helpful in the early diagnosis of the disease. Hypomelanotic macules are especially important for patients with epilepsy or babies whose number of hypomelanotic macules is more than 3. Cranial CT is of great value in the diagnosis of TS. The involvement of visceral organs such as the brain and kidney should be examined in TS patients.

Photochemotherapy inhibits angiogenesis and induces apoptosis of endothelial cells in vitro.

BACKGROUND: Photochemotherapy has long been used in the treatment of psoriasis; however, its mechanism has not been completely elucidated. Psoriasis is now regarded as an angiogenesis-related disease. Recent studies indicated that the inhibition of angiogenesis by photochemotherapy could be an underlying mechanism. It was found that photochemotherapy can downregulate the expression of angiogenic factors in keratinocytes. However, the direct effect of photochemotherapy on endothelial cells has not been studied. METHODS: In this study, we determined the effect of photochemotherapy on the proliferation of human microvascular endothelial cells through MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and cell cycle analysis. The migration assay and in vitro tube formation assay were used to investigate the migration properties and tube formation ability of human microvascular endothelial cells after psoralen plus UVA (PUVA) treatment. The apoptosis of endothelial cells elicited by photochemotherapy was also analyzed with fluorescence-activated cell sorting analysis (FACS). RESULTS: UVA (0.8-5.0 J/cm(2)) irradiation with the presence of 8-methoxypsoralen (8-MOP) (300 ng/ml) resulted in a dose-dependent reduction in the cell viabilities of endothelial cells. FACS data showed an accumulation of cells in G0/G1 phase of cell cycle and apoptotic features of cell death after UVA irradiation with psoralen. The migration properties and tube formation ability of endothelial cells were dramatically inhibited by photochemotherapy. CONCLUSION: Our results showed that photochemotherapy inhibits angiogenesis and induces apoptosis of human microvascular endothelial cells in vitro, which may be a possible mechanism of photochemotherapy in the treatment of psoriasis.