Somatic and germline mutations in the tumor suppressor gene PARK2 impair PINK1/Parkin-mediated mitophagy in lung cancer cells.

PARK2, which encodes Parkin, is a disease-causing gene for both neurodegenerative disorders and cancer. Parkin can function as a neuroprotector that plays a crucial role in the regulation of mitophagy, and germline mutations in PARK2 are associated with Parkinson's disease (PD). Intriguingly, recent studies suggest that Parkin can also function as a tumor suppressor and that somatic and germline mutations in PARK2 are associated with various human cancers, including lung cancer. However, it is presently unknown how the tumor suppressor activity of Parkin is affected by these mutations and whether it is associated with mitophagy. Herein, we show that wild-type (WT) Parkin can rapidly translocate onto mitochondria following mitochondrial damage and that Parkin promotes mitophagic clearance of mitochondria in lung cancer cells. However, lung cancer-linked mutations inhibit the mitochondrial translocation and ubiquitin-associated activity of Parkin. Among all lung cancer-linked mutants that we tested, A46T Parkin failed to translocate onto mitochondria and could not recruit downstream mitophagic regulators, including optineurin (OPTN) and TFEB, whereas N254S and R275W Parkin displayed slower mitochondrial translocation than WT Parkin. Moreover, we found that deferiprone (DFP), an iron chelator that can induce mitophagy, greatly increased the death of A46T Parkin-expressing lung cancer cells. Taken together, our results reveal a novel mitophagic mechanism in lung cancer, suggesting that lung cancer-linked mutations in PARK2 are associated with impaired mitophagy and identifying DFP as a novel therapeutic agent for PARK2-linked lung cancer and possibly other types of cancers driven by mitophagic dysregulation.

[A new Cannabis germplasm classification system and research strategies of non-psychoactive medicinal cannabis].

At present,China manages cannabis into industrial and medical types. Industrial cannabis( THC<0. 3%) is grown for fiber and seed,while medical cannabis( THC>0. 3%) is prohibited from being planted and used. In recent years,breakthroughs have been made in the applications for a variety of fields of CBD,a non-psychoactive cannabinoid of Cannabis. However,China made a minimal contribution to this international research hotspot,mainly due to the outdated way of classification on cannabis. Here we reviewed the medicinal use history of cannabis,achievements in the study of Cannabis germplasm,and first proposed the three-level classification of management: psychoactive medicinal cannabis( THC>0. 3%),non-psychoactive medicinal cannabis( THC<0. 3%,high CBD)and industrial cannabis( THC<0. 3%,for fiber and seed,low CBD). Based on summarizing the research progress of the non-psychoactive medicinal cannabis,four key research strategies of breeding,sex control,plant factory,and synthetic biology are proposed.

Rapid diagnosis of Mycobacterium marinum infection using targeted nanopore sequencing: a case report.

Mycobacterium marinum (M. marinum) is a non-tuberculous mycobacterium (NTM) that can cause infectious diseases in aquatic animals and humans. Culture-based pathogen detection is the gold standard for diagnosing NTM infection. However, this method is time-consuming and has low positivity rates for fastidious organisms. Oxford Nanopore MinION sequencing is an emerging third-generation sequencing technology that can sequence DNA or RNA directly in a culture-independent manner and offers rapid microbial identification. Further benefits include low cost, short turnaround time, long read lengths, and small equipment size. Nanopore sequencing plays a crucial role in assessing drug resistance, clinical identification of microbes, and monitoring infectious diseases. Some reports on Mycobacterium tuberculosis (MTB) using nanopore sequencing have been published, however, there are few reports on NTM, such as M. marinum. Here, we report the use of nanopore sequencing for the diagnosis of M. marinum.

Inhibition of perilipin 2 attenuates cerebral ischemia/reperfusion injury by blocking NLRP3 inflammasome activation both in vivo and in vitro.

Cerebral ischemia/reperfusion (CI/R) usually causes neuroinflammation within the central nervous system, further prompting irreversible cerebral dysfunction. Perilipin 2 (Plin2), a lipid droplet protein, has been reported to exacerbate the pathological process in different diseases, including inflammatory responses. However, the role and mechanism of Plin2 in CI/R injury are unclear. In this study, the rat models of transient middle cerebral artery occlusion followed by reperfusion (tMCAO/R) were established to mimic I/R injury, and we found that Plin2 was highly expressed in the ischemic penumbra of tMCAO/R rats. The siRNA-mediated knockdown of Plin2 significantly decreased neurological deficit scores and reduced infarct areas in rats induced by I/R. Detailed investigation showed that Plin2 deficiency alleviated inflammation of tMCAO/R rats as evidenced by reduced secretion of proinflammatory factors and the blockade of NLR family pyrin domain containing 3 (NLRP3) inflammasome activation. In vitro experiments showed that Plin2 expression was upregulated in mouse microglia subjected to oxygen-glucose deprivation/reoxygenation (OGD/R). Plin2 knockdown inhibited OGD/R-induced microglia activation and the accumulation of inflammation-related factors. Taken together, this study demonstrates that lipid droplet protein Plin2 contributes to the pathologic process of CI/R damage by impacting inflammatory response and NLRP3 inflammasome activation. Thus, Plin2 may provide a new therapeutic direction for CI/R injury.

Association of uric acid levels with cardiac syndrome X: A meta-analysis.

Objective: The pathogenesis of elevated uric acid (UA) levels in patients with cardiac syndrome X (CSX) is unclear, and the results presented in recent papers on UA levels in patients with CSX are controversial. Therefore, we conducted a meta- analysis to assess the relationship between UA levels and CSX. Methods: Three databases, including the Web of Science, EMBASE and PubMed, were systematically searched until January 2022. Fixed-effect and random-effects models were used to analyze the relationship between UA levels and CSX. Subgroup analysis and sensitivity analysis were also performed. Results: Six studies involving 406 CSX patients and 267 non-CSX were included. Our results showed a significant relationship between UA levels and CSX, with a pooled SMD of 0.68 (95% CI 0.37 to 1.00; p < 0.0001). We also found a close relationship between UA levels and CSX for patients ≥ 55 years old (SMD:0.70, 95% CI: 0.41 to 0.99, p < 0.00001), for patients < 55 years old (SMD: 0.68, 95% CI: 0.25 to 1.12, p =0 .002), for women ≥ 60% (SMD: 0.77, 95% CI: 0.33 to 1.14, p =0 .0004), for women < 60% (SMD: 0.61, 95% CI:0.23 to 0.98, p =0 .001), for BMI ≥ 28 Kg/m2 (SMD :0.61, 95% CI: 0.23 to 0.98, p =0 .001), for BMI < 28 Kg/m2 (SMD:0.75, 95% CI: 0.31 to 1.19, p =0 .0009), for publication years ≥ 2012 (SMD :0.69, 95% CI: 0.23 to 1.15, p = 0.003), for publication years < 2012 (SMD:0.73, 95% CI:0.41 to 1.05, p < 0.00001), and for Turkey (SMD:0.75, 95% CI:0.38 to 1.11, p <.0001). Sensitivity analysis showed that the pooled results remained consistent after removing any one study or converting the random-effects model to fixed-effects model. Conclusion: Our results indicated a strong association between high UA levels and CSX. However, more well-designed studies are needed to investigate whether early treatment of hyperuricemia can reduce the incidence of CSX.

HSulf-1 inhibits cell proliferation and invasion in human gastric cancer.

The HSulf-1 gene encodes an extracellular 6-O-endosulfatase and regulates the sulfation status of heparan sulfate proteoglycans (HSPG). We have demonstrated that promoter hypermethylation is correlated with the HSulf-1 silencing in gastric cancer. To investigate the functional importance of HSulf-1 silencing in gastric cancer, we restored HSulf-1 expression in the gastric cancer cell line MKN28, which lacks endogenous HSulf-1. Following restoration of expression, HSulf-1 inhibited cell proliferation, motility, and invasion in vitro, as well as significantly suppressing the MKN28 xenograft model (P < 0.05). No noticeable changes in proliferation and motility were observed following restoration of HSulf-1 in another gastric cancer cell line, namely AGS cells. Interestingly, in MKN28 cells, which have been reported to be dependent on extracellular Wnt signaling, we found that HSulf-1 inhibited the transcriptional activity of the Wnt / ß-catenin pathway and downregulated its targeted genes. Conversely, in AGS cells, in the constitutive Wnt / ß-catenin pathway is active, HSulf-1 had no effect on the activity of the Wnt / ß-catenin pathway. Furthermore, transfection of Wnt3a cDNA or ß-catenin shRNA resulted in rescue or enhancement, respectively, of the effects of HSulf-1 in MKN28 cells. Furthermore, HSPG epitope analysis confirmed that HSulf-1 affected the structure of heparan sulfate on the cell surface. Together, the results of the present study suggest that extracellular HSulf-1 may function as a negative regulator of proliferation and invasion in gastric cancer by suppressing Wnt / ß-catenin signaling at the cell surface.

Acupuncture treatment for plantar fasciitis: a randomized controlled trial with six months follow-up.

Plantar fasciitis is a common cause of heel pain. It has been suggested that some acupoints have a specific effect on heel pain. The aim of this study was to determine the efficacy and specificity of acupuncture treatment for plantar fasciitis. Subjects were randomly assigned to the treatment group (n = 28) or control group (n = 25). The treatment group received needling at the acupoint PC 7, which is purported to have a specific effect for heel pain. The control group received needling at the acupoint Hegu (LI 4), which has analgesic properties. Treatment was administered five times a week for 2 weeks, with an identical method of manual needling applied to the two acupoints. The primary outcome measure was morning pain on a 100-point visual analog scale (VAS) at one month post-treatment. Secondary outcome measures included a VAS for activity pain, overall pain rating as well as pressure pain threshold using algometry. Significant differences in reduction in pain scores, favoring the treatment group, were seen at one month for morning pain (22.6 ± 4.0 versus 12.0 ± 3.0, mean ± SEM), overall pain (20.3 ± 3.7 versus 9.5 ± 3.6) and pressure pain threshold (145.5 ± 32.9 versus -15.5 ± 39.4). No serious adverse event was observed in either group. The results indicate that acupuncture can provide pain relief to patient with plantar fasciitis, and that PC 7 is a relatively specific acupoint for heel pain.

Establishing a genomic database for the medicinal plants in the Brazilian Pharmacopoeia.

BACKGROUND: Brazil is exceptionally abundant in medicinal plant resources and has a rich ethnopharmacological history. Brazilian Pharmacopoeia (BP) acts as a national standard that regulates drug quality and has six published editions. Recent genomic approaches have led to a resurgence of interest in herbal drugs. The genomic data of plants has been used for pharmaceutical applications, protecting natural resources, and efficiently regulating the market. However, there are few genomic databases specifically for medicinal plants, and the establishment of a database that focuses on the herbs contained in the BP is urgently required. METHODS: The medicinal plant species included in each edition of the BP were analyzed to understand the evolution of the Brazilian herbal drugs. The data of 82 plants in the BP were collected and categorized into four sections: DNA barcodes, super-barcodes, genomes, and sequencing data. A typical web server architecture pattern was used to build the database and website. Furthermore, the cp-Gs of the Aloe genus in the database were analyzed as an illustration. RESULTS: A new database, the Brazilian Pharmacopoeia Genomic Database (BPGD) was constructed and is now publicly accessible. A BLAST server for species identification and sequence searching with the internal transcribed spacer 2 (ITS2), the intergenic region (psbA-trnH), and the chloroplast genome (cp-G) of Brazilian medicinal plants was also embedded in the BPGD. The database has 753 ITS2 of 76 species, 553 psbA-trnH and 190 genomes (whole genome and chloroplast genome) of 57 species. In addition, it contains 37 genome sequence data sets of 24 species and 616 transcriptome sequence data sets of 34 species and also includes 187 cp-Gs representing 57 medicinal species in the BP. Analyses of the six cp-Gs of three Aloe species identified the variable regions in the cp-Gs. These can be used to identify species and understand the intraspecific relationships. CONCLUSIONS: This study presents the first genomic database of medicinal plants listed in the latest BP. It serves as an efficient platform to obtain and analyze genomic data, accelerate studies regarding Brazilian medicinal plants and facilitate the rational development on their market regulation.

Kinetic assays of DNA polymerase fidelity: A theoretical perspective beyond Michaelis-Menten kinetics.

The high fidelity of DNA polymerase (DNAP) is critical for the faithful replication of DNA. There are several quantitative approaches to measure DNAP fidelity. Directly counting the error frequency in the replication products gives the true fidelity but it turns out very hard to implement in practice. Two biochemical kinetic approaches, the steady-state assay and the transient-state assay, were then suggested and widely adopted. In these assays, the error frequency is indirectly estimated by using kinetic theories combined with the measured apparent kinetic rates. However, whether it is equivalent to the true fidelity has never been clarified theoretically, and in particular there are different strategies using these assays to quantify the proofreading efficiency of DNAP but often lead to inconsistent results. In this paper, we make a comprehensive examination on the theoretical foundation of the two kinetic assays, based on the theory of DNAP fidelity recently proposed by us. Our studies show that while the conventional kinetic assays are generally valid to quantify the discrimination efficiency of DNAP, they are valid to quantify the proofreading efficiency of DNAP only when the kinetic parameters satisfy some constraints which will be given explicitly in this paper. These results may inspire more carefully-designed experiments to quantify DNAP fidelity.

[Floral composition and ecological distribution of wood-decaying fungi in Laojunshan National Park, Southwest China]. / ä¸½æ±Ÿè€å›å±±å›½å®¶å ¬å›­æœ¨è çœŸèŒåŒºç³»ç»„æˆä¸Žåˆ†å¸ƒç‰¹å¾.

Wood-decaying fungi are important components of forest ecosystem, mainly growing on the dead wood and decomposing lignin, cellulose, and hemi-cellulose. Understanding the ecological distribution of wood-decaying fungi are necessary to reveal their ecological function. In this study, we investigated wood-decaying fungi at three sites with different altitudes in Laojunshan National Park. Fruit bodies of fungi were collected and identified based on morphological and molecular analy-sis. In total 68 species were recorded, belonging to 40 genera, 21 families and 8 orders. Polyporaceae, Fomitopsidaceae, and Hymenochaetaceae were dominant families. The floral composition showed a distinct north temperate character, which was the most important element among all the bio-geographical elements with the highest percentage of 38.2%. These wood-decaying fungi can be divided into two ecological types: saprophytic (63 species) and ectomycohizal (5 species). Among these saprophytic fungi, 51 species could cause white rot and 12 species cause brown rot. The amounts of fungal species in deciduous broad-leaf forest (DBF), coniferous broad-leaved mixed forest (CBMF) and dark coniferous forest (DCF) were 34, 26 and 22, respectively. Postia fragilis and Stereum hirsutum, distributed in all the three types of forests, were dominant species of Laojunshan National Park.

Promoter hypermethylation correlates with the Hsulf-1 silencing in human breast and gastric cancer.

The HSulf-1 gene is an important factor that modulates the sulfation status of heparan sulfate proteoglycans (HSPGs) in the extracellular matrix, resulting in disturbance of HSPG-related signal transduction pathways. Recently, HSulf-1 has been reported to be down-regulated in several human cancers. In this study, we first cloned and characterized the 5' promoter region of the HSulf-1 gene (around 400 bp) that contained high basal promoter activity. We also found that this functional promoter region was hypermethylated in a number of human cancer cell lines. Furthermore, we found that hypermethylation in this promoter region correlated with the down-regulation of the HSulf-1 expression in human breast and gastric cancer cell lines and tissue samples. These results suggest that the promoter hypermethylation may be one of the mechanisms of the HSulf-1 gene silencing in human breast and gastric cancers. Finally, we demonstrated that the HSulf-1 promoter was more frequently (p<0.05) methylated in cell-free DNA extracted from serum samples of human breast and gastric cancer patients than that of healthy people (76.2%, 55.0% and 19.0%, respectively), indicating that detection of the HSulf-1 promoter methylation in serum samples may have clinical implications in early detection and diagnosis of human breast and gastric cancers.

Spontaneous rupture of the eyeball due to choroidal metastasis of gastric carcinoma: A case report.

RATIONALE: Although metastatic tumor is the most common intraocular malignancy, choroidal metastasis from gastric cancer is relatively rare. We present the exact course of a spontaneous rupture of the eyeball with choroidal metastasis from gastric carcinoma (GC) and the applied surgical interventions. PATIENT CONCERNS: A 59-year-old male presented with pain and vision loss on his left eye for 6 months. He was diagnosed with GC, for which he received systemic chemotherapy for a year. DIAGNOSES: Preoperative B-scan, color fundus photography, computed tomography, and magnetic resonance imaging showed a choroidal tumor in his left eye. The spontaneous rupture of the corneoscleral limbus from 2 to 5 o'clock, combined secondary glaucoma, exudative retinal detachment and choroidal detachment were found. INTERVENTIONS: The ruptured corneoscleral limbus was sutured and the orbit was lavaged with 0.4% cisplatin during the enucleation. OUTCOMES: Histopathology confirmed high homology of the choroidal metastasis and GC. He survived for 2 months after surgery, without pain or orbital neoplasms. LESSONS: Choroidal metastasis from GC rapidly progressed to spontaneous rupture of the eyeball. Careful eyeball enucleation followed by orbital lavage with chemotherapeutics may reduce metastasis risk beyond the eyeball. Additional therapeutic interventions should be considered in patients resistant to single systemic chemotherapy.

Template-specific fidelity of DNA replication with high-order neighbor effects: A first-passage approach.

DNA replication fidelity is a critical issue in molecular biology. Biochemical experiments have provided key insights on the mechanism of fidelity control by DNA polymerases in the past decades, whereas systematic theoretical studies on this issue began only recently. Because of the underlying difficulties of mathematical treatment, comprehensive surveys on the template-specific replication kinetics are still rare. Here we propose a first-passage approach to address this problem, in particular the positional fidelity, for complicated processes with high-order neighbor effects. Under biologically relevant conditions, we derived approximate analytical expressions of the positional fidelity which show intuitively how some key kinetic pathways are coordinated to guarantee the high fidelity, as well as the high velocity, of the replication processes. It is also shown that the fidelity at any template position is dominantly determined by the nearest-neighbor template sequences, which is consistent with the idea that replication mutations are randomly distributed in the genome.

Effects of Adipose-derived Mesenchymal Stem Cell Exosomes on Corneal Stromal Fibroblast Viability and Extracellular Matrix Synthesis.

BACKGROUND: Corneal stromal cells (CSCs) are components of the corneal endothelial microenvironment that can be induced to form a functional tissue-engineered corneal endothelium. Adipose-derived mesenchymal stem cells (ADSCs) have been reported as an important component of regenerative medicine and cell therapy for corneal stromal damage. We have demonstrated that the treatment with ADSCs leads to phenotypic changes in CSCs in vitro. However, the underlying mechanisms of such ADSC-induced changes in CSCs remain unclear. METHODS: ADSCs and CSCs were isolated from New Zealand white rabbits and cultured in vitro. An Exosome Isolation Kit, Western blotting, and nanoparticle tracking analysis (NTA) were used to isolate and confirm the exosomes from ADSC culture medium. Meanwhile, the optimal exosome concentration and treatment time were selected. Cell Counting Kit-8 and annexin V-fluorescein isothiocyanate/propidium iodide assays were used to assess the effect of ADSC- derived exosomes on the proliferation and apoptosis of CSCs. To evaluate the effects of ADSC- derived exosomes on CSC invasion activity, Western blotting was used to detect the expression of matrix metalloproteinases (MMPs) and collagens. RESULTS:: ADSCs and CSCs were successfully isolated from New Zealand rabbits. The optimal concentration and treatment time of exosomes for the following study were 100 µg/ml and 96 h, respectively. NTA revealed that the ADSC-derived exosomes appeared as nanoparticles (40-200 nm), and Western blotting confirmed positive expression of CD9, CD81, flotillin-1, and HSP70 versus ADSC cytoplasmic proteins (all P < 0.01). ADSC-derived exosomes (50 µg/ml and 100 µg/ml) significantly promoted proliferation and inhibited apoptosis (mainly early apoptosis) of CSCs versus non-exosome-treated CSCs (all P < 0.05). Interestingly, MMPs were downregulated and extracellular matrix (ECM)-related proteins including collagens and fibronectin were upregulated in the exosome-treated CSCs versus non-exosome-treated CSCs (MMP1: t = 80.103, P < 0.01; MMP2: t = 114.778, P < 0.01; MMP3: t = 56.208, P < 0.01; and MMP9: t = 60.617, P < 0.01; collagen I: t = -82.742, P < 0.01; collagen II: t = -72.818, P < 0.01; collagen III: t = -104.452, P < 0.01; collagen IV: t = -133.426, P < 0.01, and collagen V: t = -294.019, P < 0.01; and fibronectin: t = -92.491, P < 0.01, respectively). CONCLUSION:: The findings indicate that ADSCs might play an important role in CSC viability regulation and ECM remodeling, partially through the secretion of exosomes.

Cell viability and extracellular matrix synthesis in a co-culture system of corneal stromal cells and adipose-derived mesenchymal stem cells.

AIM: To investigate the impact of adipose-derived mesenchymal stem cells (ADSCs) on cell viability and extracellular matrix (ECM) synthesis of corneal stromal cells (CSCs). METHODS: ADSCs and CSCs were obtained from the corneas of New Zealand white rabbits and indirectly co-cultured in vitro. The proliferative capacity of CSCs in the different groups was assessed by CCK-8 assays. Annexin V-fluorescein isothiocyanate (FITC)/proliferation indices (PI) assays were used to detect the apoptosis of CSCs. The expression levels of matrix metalloproteinase (MMP), such as MMP1, MMP2, MMP9, and collagens were also evaluated by Western blot. RESULTS: ADSCs significantly promoted proliferation and invasion of CSCs in the indirect co-culture assays. The co-cultural group displayed much higher ability of proliferation, especially under the co-culture conditions of ADSCs for 3d, compared with that CSCs cultured alone. The PI of CSCs in the co-culture system were increased approximately 3-8-fold compared with the control group. A significant change was observed in the proportions of cells at apoptosis (early and late) between the negative control group (6.34% and 2.06%) and the ADCSs-treated group (4.69% and 1.59%). The expression levels of MMPs were down regulated in the co-culture models. Compared with the control group, the decrease intensities of MMP-1, MMP-2 and MMP-9 in CSCs/ADSCs group were observed, 3.90-fold, 1.09-fold and 3.03-fold, respectively. However, the increase intensities of collagen type (I, II, III, IV, and V) in CSCs were observed in CSCs/ADSCs group, 3.47-fold, 4.30-fold, 2.35-fold, 2.55-fold and 2.43-fold, respectively, compared to that in the control group. The expressions of aldehyde dehydrogenase and fibronectin in CSCs were upregulated in the co-culture models. CONCLUSION: ADSCs play a promotive role in CSCs' growth and invasion, which may be partially associated with MMPs decrease and collagens increase, resulting in a positive participation in the plasticity and ECM synthesis of CSCs. This provided a new insight into the extensive role of ADSCs in CSCs and a potential molecular target for corneal therapy.

[Effect of scalp acupuncture on glucose metabolism in brain of patients with depression].

OBJECTIVE: To observe the effect of scalp acupuncture (SA) on the glucose metabolism in different regions of brain in patients with depression by positron emission computed tomography (PET). METHODS: Twelve depressive patients were treated by scalp acupuncture on middle line of vertex (MS5), middle line of forehead (MS1) and bilateral lateral line 1 of forehead (MS2), once a day for six days per week, and received PET detection on different region of brain before and after 6 weeks acupuncture treatment. Semiquantitative analysis was used to compare the average values of radioactive count gotten from various brain regions before and after treatment, which could reflect the condition of glucose metabolism at the brain region detected. RESULTS: SA could increase the glucose metabolism at bilateral frontal lobes, bilateral parietal lobes, right occipital lobe, right caudate nucleus, right cingulated gyrus and left cerebellum and decrease that at right temporal lobe and bilateral thalamus. CONCLUSION: SA on MS5, MS1 and MS2 in depressive patients could influence the glucose metabolism in various brain regions. It primarily illustrated that the mechanism of SA in treating depression is related with its regulation on cortex-limbic circuitry dysfunction and increase the glucose metabolism in various brain regions.

Aberrant Wnt/ß-catenin signaling and elevated expression of stem cell proteins are associated with osteosarcoma side population cells of high tumorigenicity.

According to the cancer stem cell theory, the presence of a small sub­population of cancer cells, termed cancer stem cells (CSCs), have a significant implication on cancer treatment and are responsible for tumor recurrence. Previous studies have reported that alterations in the Wnt/ß­catenin signaling are crucial in the maintenance of CSCs. In the present study, the characteristic features and activation of Wnt/ß­catenin signaling in CSCs from osteosarcoma, an aggressive human bone tumor, were investigated. In total, ~2.1% of the cancer stem­like side population (SP) cells were identified in the osteosarcoma samples. The results of subsequent western blot and reverse transcription­quantitative polymerase chain reaction analyses revealed that the protein levels of ß­catenin and cyclin D1 were markedly upregulated in the fluorescence­activated cell sorted osteosarcoma SP cells. In addition, the elevated expression levels of stem cell proteins, including CD133, nestin Oct­4, Sox­2 and Nanog were significantly higher in the SP cells, which contributed to self­renewal and enhanced the proliferation rate of the SP cells. Furthermore, the SP cells were found to be highly invasive and able to form tumors in vivo. Taken together, these data suggested that the identification of novel anticancer drugs, which suppress the Wnt/ß­catenin signaling and its downstream pathway may assist in eradicating osteosarcoma stem cells.

Effects of sleep deprivation on gamma-amino-butyric acid and glutamate contents in rat brain.

OBJECTIVE: To investigate the effects of sleep deprivation on the contents of gamma-amino-butyric acid (GABA) and glutamate (GLU) in rat brain. METHODS: Thirty rats were assigned into sleep-deprivation (SD, n=10) group (subjected to treatment on small platforms 6.5 cm of diameter), tank control (TC, n=10) group (treated on large platforms 14 cm of diameter) and control group kept in cages (CC, n=10). Rats in SD group were deprived of rapid eye movement sleep for 96 h by "flower pot" technique. High-performance liquid chromatography was used to determine the GABA and GLU contents in the 3 brain regions of the rats: the frontal cortex, hypothalamus and brain stem. RESULTS: The rats in SD group showed an increase in the contents of GABA and lowered ratio of GLU/GABA in all the 3 brain regions (P<0.01) compared with the rats in CC group; GLU contents were also increased in the brain stem and hypothalamus of the rats in SD group (P<0.05). No significant changes were found in TC group. CONCLUSION: Rapid eye movement sleep deprivation induces significant changes in the contents and proportion of amino-acid neurotransmitter in rat brain.

[Effect of Tongxinluo on the proliferation and differentiation of rat embryonic neural stem cells].

OBJECTIVE: To investigate the effect of Tongxinluo in on the proliferation and differentiation of rat embryonic neural stem cells (NSCs). METHODS: NSCs were isolated from 12- to 14-day SD rat embryo and treated with Tongxinluo at different doses, and the proliferation and differentiation of the cells were observed by immunofluorescence staining at different time points. RESULTS: The ratio of embryonic NSCs labeled with nestin decreased soon after Tongxinluo treatment, but increased afterwards. Significant difference was noted in the number of cells labeled with beta-tubulin between Tongxinluo group and the control group 3 and 7 days after the treatment, and also between high-dose and low-dose Tongxinluo groups at 7 days. CONCLUSION: Tongxinluo can induce the proliferation and neuronal differentiation of rat embryonic NSCs, and the effect is related to the dose of Tongxinluo administered.

[18FDG PET cerebral function imaging in 10 vascular dementia patients receiving needling at Baihui(DU20), Shuigou(DU26) and Shenmen(HT7)].

OBJECTIVE: To observe the effect of needling at Baihui(DU20), Shuigou(DU26) and Shenmen(HT7) on glucose metabolism in different regions of the brain in vascular dementia patients. METHODS: Ten patients with vascular dementia were randomized into 2 groups, and the patients in the control group received the treatment with needling in the routine acupoints for hemiplegia (chosen from 6 Yang meridians of the hand and foot), while those in treatment group received needling at 3 additional acupoints, namely Baihui(DU20), Shuigou(DU26) and Shenmen(HT7), which were specific for dementia treatment. All the patients were examined by positron-emission tomography(PET) to detect the glucose metabolism in different brain regions before and after acupuncture treatment by means of semiquantitative analysis of the average values of radioactive count in the brain regions. RESULTS: Needling at the additional 3 acupoints could obviously enhance glucose metabolism in the bilateral frontal lobes, bilateral thalamus, temporal lobe and lentiform nucleus on the uncompromised side. CONCLUSION: The effect of needling at Baihui(DU20), Shuigou(DU26) and Shenmen(HT7) on vascular dementia is closely related to improved cerebral glucose metabolism.