Anti-Melanoma Activity of Single Intratumoral Injection of ZnPc Micelles Mixed With in situ Gel in B16 Bearing Mouse.

Photodynamic therapy (PDT) is a potential treatment strategy for melanoma. As a second-generation photosensitizer, Zinc phthalocyanine (ZnPc) has many advantages for anti-tumor PDTs, such as strong absorption in the red and near infrared regions, high photo and chemical stability, etc. However, ZnPc has a poor water solubility and is apt to aggregate due to the π-π interaction between molecules, which limits its applications. In this study, various solvents and surfactants were screened for dissolving ZnPc and preparing ZnPc@SDC-TPGS micelle and thermosensitive in situ gel. After the cytotoxic effects of thermosensitive gels on PDT were tested, the antitumor effects on PDT of them in mice by intratumoral injection were evaluated, including body weight, and tumor weight, volume and morphology. The cell death pathway and the relationship of reactive oxygen species yield with apoptotic rate of tumor cells induced by ZnPc in situ gel were investigated. The results were that N-methyl-pyrrolidone (NMP) mixed with 2 % SDC and aqueous solution containing 2 % TPGS and 2 % SDC were used to synthesize ZnPc@SDC-TPGS micelle and the thermosensitive in situ gel. The cytotoxic effects of thermosensitive gels showed good tumor suppression of ZnPc@SDC-TPGS in situ gel and no toxicity of the blank gel. Intratumoral injection in situ gel containing 3 µg ZnPc under irradiation demonstrated good tumor inhibition in mice with melanoma. Apoptosis has been established as the primary pathway of cell death, and the production of reactive oxygen species (ROS) plays a crucial role in cellular apoptosis induced by ZnPc@SDC-TPGS in situ gel. In conclusion, the intratumoral injection of ZnPc@SDC-TPGS thermosensitive in situ gel provides a promising local treatment option for melanoma.

Impacts of Eucalyptus plantation on soil and water losses in a typical small watershed in mountainous area of southern China.

Unreasonable exploitation of artificial forest causes severe soil erosion in the mountainous areas of sou-thern China. The spatial-temporal variations of soil erosion in typical small watershed with artificial forest has signifi-cant implications for artificial forest exploitation and sustainable development of mountainous ecological environment. In this study, we used revised universal soil loss equation (RUSLE) and geographic information system (GIS) to evaluate the spatial and temporal variations of soil erosion and its key drivers of Dadingshan watershed in mountainous area of western Guangdong. The results showed that the erosion modulus was 1948.1 t·km-2·a-1 (belonging to light erosion) in the Dadingshan watershed. However, the spatial variation of soil erosion was substantial, with variation coefficient of 5.12. The maximal soil erosion modulus was 191127 t·km-2·a-1. Slight erosion (<500 t·km-2·a-1) accounted for 80.6% of the total watershed area. The moderate erosion and above (>2500 t·km-2·a-1) were mainly distributed in young Eucalyptus forest area with less than 30% of the vegetation coverage, which contributed nearly 75.7% of total soil erosion. During 2014-2019, the interannual variations of mean erosion of Dadingshan catchment was modest, but the spatial variation of soil erosion was large. Vegetation cover, slope, and rainfall were key drivers of such variation. The destruction of natural vegetation resulted by plantation exploitation was the primary cause of soil erosion in afforestation areas. Soil erosion significantly increased with the increases of slope gradient in the young forest area, which was aggravated by extreme rainfall. However, soil erosion gradually decreased with the increases of the age of Eucalypt plantation. Therefore, the hot spot of soil erosion was young forest areas of Eucalypt plantation with slope >25°, and the key period for soil erosion control was the first 2-3 years after Eucalyptus planting. We suggested that reasonable afforestation measures should be used in area with >25° slopes, and that the destruction of natural vegetation should be avoided on hillslope with >35° slope gradient. The road construction standards and forest management should be further improved to address the challenge of extreme rainfalls.

Effects of common afforestation tree species on soil bacterial community and microbial functional guilds in subtropical forests.

To understand the effects of common afforestation tree species on soil microbial community in subtropical forests, seven different tree species were selected as the research object, including Pinus massoniana, Mytilaria laosensis, Liquidambar formosana, Ilex chinensis, Michelia macclurei, Quercus acutissima and Betula luminifera. Based on 16S rRNA high-throughput sequencing and real-time quantitative PCR techniques, we explored the effects of different tree species on soil bacterial community composition, diversity and microbial functional guilds. The results showed that Acidobacteria, Proteobacteria, and Actinobacteria were the dominant bacterial phyla, and that there was no significant difference in bacterial diversity or richness index among different tree species. Results of redundancy analysis suggested that soil bulk density, soil C/N, litter nitrogen content, and litter C/N were the predominant factors determining soil bacterial community composition. The afforestation tree species had significant effects on functional gene abundances of ammonia oxidizing archaea, ammonia oxidizing bacteria and complete ammonia oxidation. Comammox were dominant in abundance. Ammonia oxidizing archaea amoA gene was the only type whose abundance showed significant correlation with soil nitrate content, suggesting that ammonia oxidizing archaea could play a dominant role in the autotrophic nitrification in the acidic subtropical forest soils. The afforestation tree species had significant effects on functional gene abundances of ammonia oxidizing microorganisms. Results of correlation analysis showed that litter nitrogen content was the driving factor for the abundance of ammonia oxidizing microorganisms. Our study provided strong evidence that the responses of soil microbial functional guilds to tree species were more sensitive than bacterial community composition. Future studies should explore the mechanisms of tree plantations on forest ecosystem functioning from the perspective of microbial functional guilds.

Efficacy of acupuncture at ghost points combined with fluoxetine in treating depression: A randomized study.

BACKGROUND: Depression affects more than 350 million people worldwide. In China, 4.2% (54 million people) of the total population suffers from depression. Psychotherapy has been shown to change cognition, improve personality, and enhance the ability to cope with difficulties and setbacks. While pharmacotherapy can reduce symptoms, it is also associated with adverse reactions and relapse after drug withdrawal. Therefore, there has been an increasing emphasis placed on the use of non-pharmacological therapies for depression. The hypothesis of this study was that acupuncture at ghost points combined with fluoxetine would be more effective than fluoxetine alone for the treatment of depression. AIM: To investigate the efficacy of acupuncture at ghost points combined with fluoxetine for the treatment of patients with depression. METHODS: This randomized controlled trial included patients with mild to moderate depression (n = 160). Patients received either acupuncture at ghost points combined with fluoxetine (n = 80) or fluoxetine alone (control group, n = 80). Needles were retained in place for 30 min, 5 times a week; three treatment cycles were administered. The Mann-Whitney U test was used to compare functional magnet resonance imaging parameters, Hamilton depression rating scale (HAMD) scores, and self-rating depression scale (SDS) scores between the acupuncture group and control group. RESULTS: There were no significant differences in HAMD or SDS scores between the acupuncture group and control group, before or after 4 wk of treatment. The acupuncture group exhibited significantly lower HAMD and SDS scores than the control group after 8 wk of treatment (P < 0.05). The acupuncture group had significantly lower fractional Amplitude of Low Frequency Fluctuations values for the left anterior wedge leaf, left posterior cingulate gyrus, left middle occipital gyrus, and left inferior occipital gyrus after 8 wk. The acupuncture group also had significantly higher values for the right inferior frontal gyrus, right insula, and right hippocampus (P < 0.05). After 8 wk of treatment, the effective rates of the acupuncture and control groups were 51.25% and 36.25%, respectively (P < 0.05). CONCLUSION: The study results suggest that acupuncture at ghost points combined with fluoxetine is more effective than fluoxetine alone for the treatment of patients with mild to moderate depression.

[Clinical observation of different surgical methods for metaphyseal sequelae of ulna].

OBJECTIVE: To investigate the effect and safety of ulnar osteochondroma resection, ulnar minimally invasive osteotomy, external fixation and ulnar lengthening in the treatment of forearm deformity of metaphyseal extension of ulna. METHODS: From August 2005 to December 2013, there were 20 cases of ulnar metaphyseal sequelae, including 15 males and 5 females, aged from 7 to 13(10.00±2.34) years, the course of disease ranged for 6 to 11(8.10±1.52) months. The clinical manifestations were shortening of the affected forearm and bending to the ulnar side. The postoperative evaluation included pain, activities of daily living, orthopedic effect and the range of motion of wrist, elbow and forearm. The radiological evaluation included ulnar length, radial joint inclination angle and wrist epiphysis growth. RESULTS: All patients healed without infection. The only operation related to complications was ulnar lengthening, including 1 case of nonunion, 2 cases of ulnar lengthening callus fracture and 1 case of temporary radial nerve palsy. All patients were followed up for 4 to 7.5 years, with an average of (6.03±1.33) years. There were statistically significant differences in changes of wrist radial deviation, ulnar deviation, forearm pronation and supination in all cases (P<0.05). Radiological evaluation parameters (ulnar variance, radial joint angle, wrist slip) were improved, and the differences were statistically significant(P<0.05). The modified Green and O'Brien wrist function scores at the last follow-up were significantly different from those before operation (P<0.05). The clinical effect of wrist joint was significantly different from that before operation (P<0.05). The Mayo elbow function scoreat the latest follow-up was significantly different from that before operation (P<0.05). The clinical effect of elbow joint was significantly different from that before operation (P<0.05). CONCLUSION: Ulnar lengthening is not beneficial to prevent the development of long-term deformity. Simple resection of osteochondroma of distal ulna is beneficial to prevent the development of deformity. Patients with limited rotation of wrist joint and forearm and strong demand for improvement of appearance can be actively treated.

Enhanced Antitumor Efficacy of Curcumin-Loaded PLGA Nanoparticles Coated with Unique Fungal Hydrophobin.

Modifications to the surface chemistry, charge, and hydrophilicity/hydrophobicity of nanoparticles are applicable approaches to the alterations of the in vivo fate of intravenously administered nano-sized drug carriers. The objective of this study is to investigate the in vitro and in vivo antitumor efficacies of curcumin PLGA nanoparticles in relation to their surface structural modification via self-assembling coating with unique fungal hydrophobin. The hydophobin-coated curcumin PLGA nanoparticles (HPB PLGA NPs) were obtained by simply soaking curcumin-loaded PLGA nanoparticles (PLGA NPs) in aqueous fungal hydrophobin solution. The in vitro drug release behavior of the HPB PLGA NPS was also tested. The cytotoxicity and cellular uptake of these nanoparticles were determined in HepG2, A549, and Hela cell lines using MTT assay method and CLSM observation. The in vivo antitumor activity was evaluated in Hela tumor xenografted mice model. Compared with the PLGA NPs, the size and zeta potential of the nanoparticles were changed after hydrophobin coating, whereas similar in vitro release pattern was observed. The pharmacodynamics study showed prolonged blood retention of both nano-formulations than that of free curcumin, but no significant difference between the hydrophobin coated and uncoated nanoparticles. It was found that HPB PLGA NPs had increased cytotoxicities, higher cellular uptake, and improved antitumor efficacy. Surface modification of nanoparticles via self-assembling of hydrophobin is a convenient and promising method of changing particle surface physiochemical properties and antitumor performances. Further investigations, especially on tissue distribution, were needed to assess the potential application of the hydrophobin self-assembling coating in nano-drug delivery carriers.

Mangiferin ameliorates insulin resistance by inhibiting inflammation and regulatiing adipokine expression in adipocytes under hypoxic condition.

Adipose tissue hypoxia has been recognized as the initiation of insulin resistance syndromes. The aim of the present study was to investigate the effects of mangiferin on the insulin signaling pathway and explore whether mangiferin could ameliorate insulin resistance caused by hypoxia in adipose tissue. Differentiated 3T3-L1 adipocytes were incubated under normal and hypoxic conditions, respectively. Protein expressions were analyzed by Western blotting. Inflammatory cytokines and HIF-1-dependent genes were tested by ELISA and q-PCR, respectively. The glucose uptake was detected by fluorescence microscopy. HIF-1α was abundantly expressed during 8 h of hypoxic incubation. Inflammatory reaction was activated by up-regulated NF-κB phosphorylation and released cytokines like IL-6 and TNF-α. Glucose uptake was inhibited and insulin signaling pathway was damaged as well. Mangiferin substantially inhibited the expression of HIF-1α. Lactate acid and lipolysis, products released by glycometabolism and lipolysis, were also inhibited. The expression of inflammatory cytokines was significantly reduced and the damaged insulin signaling pathway was restored to proper functional level. The glucose uptake of hypoxic adipocytes was promoted and the dysfunction of adipocytes was relieved. These results showed that mangiferin could not only improve the damaged insulin signaling pathway in hypoxic adipocytes, but also ameliorate inflammatory reaction and insulin resistance caused by hypoxia.

Synthesis of 1,2,3-Thiadiazole and Thiazole-Based Strobilurins as Potent Fungicide Candidates.

Strobilurin fungicides play a crucial role in protecting plants against different pathogens and securing food supplies. A series of 1,2,3-thiadiazole and thiazole-based strobilurins were rationally designed, synthesized, characterized, and tested against various fungi. Introduction of 1,2,3-thiadiazole greatly improved the fungicidal activity of the target molecules. Compounds 8a, 8c, 8d, and 10i exhibited a relatively broad spectrum of fungicidal activity. Compound 8a showed excellent activities against Gibberella zeae, Sclerotinia sclerotiorum, and Rhizoctonia cerealis with median effective concentrations (EC50) of 2.68, 0.44, and 0.01 µg/mL, respectively; it was much more active than positive controls enestroburin, kresoxim-methyl, and azoxystrobin with EC50 between 0.06 and 15.12 µg/mL. Comparable or better fungicidal efficacy of compound 8a compared with azoxystrobin and trifloxystrobin against Sphaerotheca fuliginea and Pseudoperonspera cubensis was validated in cucumber fields at the same application dosages. Therefore, compound 8a is a promising fungicidal candidate worthy of further development.

Improvement of the Antitumor Efficacy of Intratumoral Administration of Cucurbitacin Poly(Lactic-co-Glycolic Acid) Microspheres Incorporated in In Situ-Forming Sucrose Acetate Isobutyrate Depots.

Localized drug delivery strategies for cancer therapy have been introduced for decades as a means of increasing drug concentration at tumor target site and minimizing systemic toxicities. In this paper, a combination of microspheres (MSs) and sucrose acetate isobutyrate (SAIB) in situ-forming implants (ISFIs) was evaluated for improving antitumor efficacy via intratumoral injection. Monodispersed cucurbitacin (Cuc)-loaded Poly (lactic-co-glycolic acid) (PLGA) MSs with mean diameter of about 5 µm were fabricated by Shirasu porous Glass (SPG) membrane emulsification technique, and their properties were investigated. The in vitro drug release pattern, antimelanoma efficiency, and drug distribution in tumor of three different intratumoral injection systems, that is, MSs, SAIB ISFIs, and combination of MSs and SAIB ISFIs (SAIB-MSs), was investigated. The Cuc-loaded MSs prepared by PLGA (LA/GA = 50:50, inherent viscosity = 0.87 dL/g), has an appropriate release pattern with lower initial burst and delayed drug release. SAIB-MSs have a much slower drug release rate than that of MSs or SAIB ISFIs. SAIB-MSs showed the best antitumor efficacy in melanoma-bearing mice model, and the results of drug distribution in tumor revealed that the incorporation MSs in SAIB solution obviously extended the residence of drug in tumor. The low Cuc concentration in tumor periphery region after intratumoral administration of SAIB-MSs demonstrated poor drug penetration of this system. For further improving the antitumor efficacy of intratumoral chemotherapy, elegant designing to carriers with both extended residency and wide drug distribution in tumor is needed.

UPLC-MS/MS-ESI assay for simultaneous determination of magnolol and honokiol in rat plasma: application to pharmacokinetic study after administration emulsion of the isomer.

ETHNOPHARMACOLOGICAL RELEVANCE: Magnolia officinalis is one of the commonly used in traditional Chinese medicine for the treatment of fever, chronic bronchitis and stomach ailments. Magnolol and honokiol are isomers with hydroxylated biphenol compound in the extract of Magnolia officinalis. This study aims to determine the isomers in rat plasma and evaluate their pharmacokinetic pattern after administration emulsion. MATERIALS AND METHODS: Sprague Dawley male rats received either an intravenous (i.v.25, mg/kg) or oral (50mg/kg) dose of the emulsion of the isomer. A sensitive and specific ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method was developed for the investigation of the pharmacokinetics of magnolol and honokiol in rats. Kaempferol was employed as an internal standard. RESULTS: The plasma samples were deproteinized with acetonitrile, the post-treatment samples were analyzed on an Agela C18 column interfaced with a triple quadrupole tandem mass spectrometer in negative electrospray ionization mode. Acetonitrile and 5 mmol/L ammonium acetate buffer solution (65: 35, v/v) was used as the mobile phase at a flow rate of 0.2 mL/min. Following oral administration of emulsion to rats, magnolol attained mean peak plasma concentrations of 426.4 ± 273.8 ng/mL at 1.20 h, whereas honokiol reached peak plasma concentrations of 40.3 ± 30.8 ng/mL at 0.45 h. The absolute bioavailability of magnolol and honokiol is 17.5 ± 9.7% and 5.3 ± 11.7%. By comparison, the AUC0-∞ of magnolol was 5.4 times higher than that of honokiol after intravenous administration, but AUC0-∞ of magnolol was about 18-fold higher than honokiol after oral administration.

Multicentric reticulohistiocytosis associated with liver carcinoma: report of a case.

We report a unique case of multicentric reticulohistiocytosis (MRH) associated with liver carcinoma. A 61-year-old man presented with a 4-month history of nonpruritic, generalized, ruby-red papules and nodules, accompanied by fever, joint swelling and difficulty in swallowing. Skin histology showed polymorphic histiocyte infiltration with typical 'ground glass' cytoplasm. Further immunohistochemical studies characterized the lesions as positive for leukocyte common antigen, HLA-DR and CD68. The patient had a history of hepatitis B, and systemic examination, including carcinoma index and type-B ultrasonic examination, revealed high levels of AFP and a solid tumor, which was considered malignant, localized on the right lobe of the liver. Treatment of the liver carcinoma resulted in a significant improvement of the skin symptoms. This is the first case study to report an association between MRH and liver carcinoma. A review of the English-language literature reveals the close linkage between MRH and malignancy. All patients with MRH should be evaluated and monitored carefully to determine the underlying neoplasm.

Gene expression profile and overexpression of apoptosis-related genes (NGFI-B and Gadd 45 gamma) in early phase of Thy-1 nephritis model.

Mesangioproliferative glomerulonephritis (MPGN) is a disease of high incidence in humans. Rat Thy-1 nephritis (Thy-1 N), namely, anti-thymocyte serum (ATS)-induced nephritis, is considered to be an animal model for studying MPGN. Although previous studies have demonstrated that glomerular mesangial cell (GMCs) injury might be a feature of Thy-1 N, the mechanism of the disease (i.e., GMC apoptosis) remains unclear. We have examined the pathologic changes of GMCs and the gene expression profile of renal tissues in Thy-1 N. The pathologic changes of Thy-1 N include three phages: GMC apoptosis (40 min), necrosis (2 h), and proliferation (5 days). Many TUNEL-positive cells are found 40 min after administration of ATS. Concomitantly, 341 genes are up-regulated, whereas 392 genes are down-regulated, as shown by microarrays analysis. The mRNA and protein of two of the up-regulated genes (nerve growth factor induced protein I-B, NGFI-B; growth arrest- and DNA-damage-inducible protein 45 gamma, Gadd 45 gamma) in the GMC apoptotic phase of Thy-1 N are markedly elevated, as observed by real-time polymerase chain reaction and immunohistochemistry. Our data indicate that pathologic changes of Thy-1 N are involved in the abnormal gene profile. The overexpression of the NGFI-B and Gadd 45 gamma genes may be associated with GMC apoptosis of Thy-1 N.

[Determination of hydromorphone in Beagle dogs plasma by liquid chromatography-tandem mass spectrometry].

AIM: To establish a LC/MS/MS method for determination of hydromorphone (HYD) in Beagle dog plasma. METHODS: After incubation with beta-glucuronidase for 16 h, an aliquot of 0.1 mL plasma was treated by liquid-liquid extraction. The analytes of interest were separated on a Zorbax SB C8 column with the mobile phase consisting of methanol-water- formic acid (65: 35: 1). Atmospheric pressure chemical ionization source of MS was applied and operated in positive ion mode. RESULTS: The linear calibration curve was obtained in the concentration range of 0.80 - 200.0 microg x L(-1). The lower limit of quantification was 0.80 microg x L(-1). The inter-day and intra-day precision (RSD) was below 6.0%, and the accuracy (RE) was within 1% calculated from QC samples. The method was used to determine the pharmacokinetic parameters of HYD after a single oral administration of 4 mg HYD sustained release tablets to Beagle dogs. CONCLUSION: The method was proved to be specific, sensitive, and suitable for the pharmacokinetic study of HYD sustained release formulation.

Pharmacokinetics of 9-nitro-20(S)-camptothecin in rats.

AIM: To study the pharmacokinetics and the excretion of 9-nitro-20(S)-camptothecin (9-NC) in rats. METHODS: Each rat was given a single dose at random by iv or ig administration. Serial plasma and excreta samples were collected and the pharmacokinetic behavior of 9-NC in rats was characterized by specific liquid chromatographic assays. Individual 9-NC plasma-concentration data were analyzed by both noncompartmental and compartmental analysis. For dose proportionality, AUC- and Cmax-dose relationships were evaluated by linear regression, and t1/2 and CLtot were compared by an analysis of variance model. Also, the excretion of the parent drug was estimated. RESULTS: After iv administration of 9-NC at the doses of 1.5, 3, and 6 mg/kg, the t1/2 values for 9-NC were estimated to be 0.5, 0.5, and 0.7 h, respectively, and the mean AUC0-t values were 633, 1606, and 3011 h.microg.L(-1), respectively. 9-NC was rapidly absorbed, reaching mean Cmax of 203, 417, and 1150 microg/L at Tmax of 0.3, 0.2, and 0.3 h at the doses of 3, 6, and 12 mg/kg, respectively. The mean AUC0-t values were 269, 439, and 881 h.microg.L(-1), and the mean t1/2 values were 1.7, 0.9, and 0.9 h, respectively. The absolute oral bioavailability of 9-NC was calculated to be 14.6 %, which was consistent with the ratio of the total cumulative excretion in the urine and bile by ig to that by iv injection. CONCLUSION: The kinetic process of 9-NC in rats in vivo was best fitted to a two-compartmental model. For iv administration, the pharmacokinetics are not dose-dependent. The oral bioavailability of 9-NC was low. Renal excretion was the primary elimination route of the parent drug after iv administration, however, after ig administration the unchanged drug was largely excreted in the feces because of the poor absorption.