Mechanisms of acupuncture for primary dysmenorrhea based on PI3K/Akt/mTOR signaling pathway in rats. / 基于PI3K/Akt/mTORä¿¡å·é€šè·¯æŽ¢è®¨ç”µé’ˆæ²»ç–—åŽŸå‘æ€§ç—›ç»å¤§é¼ çš„æœºåˆ¶.

OBJECTIVES: To observe the effect of electroacupuncture(EA) on phosphatidylinositol-3-kinases(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) signaling pathway of uterus tissue in rats with primary dysmenorrhea(PDM), so as to investigate its mechanisms underlying improvement of PDM. METHODS: Thirty healthy non-pregnant female SD rats were randomly divided into blank, model and EA groups, with 10 rats in each group. The PDM model was established by subcutaneous injection of estradiol diphenhydrate combined with intraperitoneal injection of oxytocin. For rats of the EA group, EA(50 Hz, a tolerable current intensity) was applied to "Guanyuan"(CV4) and bilateral "Sanyinjiao"(SP6) for 20 min, once a day for 10 consecutive days. The number of writhing, wri-thing score, and writhing latency were observed. The uterine histopathological changes were observed by H.E. staining, and the ultrastructural changes of uterine tissue cells in each group were observed by transmission electron microscopy. The contents of prostaglandin E2(PGE2), prostaglandin F2α(PGF2α) and ratios of PGF2α/PGE2 in the serum and uterine tissue were detected by ELISA. The relative expression levels of PI3K, Akt and mTOR and their phosphorylation proteins in the uterine tissue were detected by Western blot and the ratios were calculated. RESULTS: Compared with the blank group, the number and score of writhing, latency of writhing, pathological injury score, contents of PGF2α and ratios of PGF2α/PGE2 in the serum and uterine tissue, and the levels of p-PI3K/PI3K, p-Akt/Akt and p-mTOR/mTOR in the uterine tissue were significantly increased in the model group(P<0.01, P<0.05), while contents of PGE2 in the serum and uterine tissue were reduced(P<0.05). In comparison with the model group, the number of writhing and writhing score, pathological injury score, contents of PGF2α and ratios of PGF2α/PGE2 in both the serum and uterine tissue, the levels of p-PI3K/PI3K, p-Akt/Akt and p-mTOR/mTOR were obviously decreased(P<0.05, P<0.01), whereas the writhing latency was considerably prolonged in the EA group(P<0.01), with elevated contents of PGE2 in the serum and uterine tissue(P<0.05). H.E. staining showed slight dilation of uterine glandular cavity, and severe endometrial edema with extensive cell shedding and a large number of vacuole-like degeneration, apoptosis, pyknosis or fragmentation or disappearance of the nucleus, and neutrophil infiltration in the model group, which were relatively milder in the EA group. Ultrastructural results showed irregular fibroblasts of uterine tissue cells, obvious cytoplasmic edema, reduction in cytoplasmic electron density, seriously irregular nuclei, severe edema of mitochondria with dissolved matrix, fracture and disappearance of mitochondrial crests and vacuolation, and moderate dilation of rough endoplasmic reticulum in the model group, which were milder in the EA group. CONCLUSIONS: EA can improve pain and uterine inflammatory response in PDM rats, which may be associated with its functions in reducing uterine PGF2α and down-regulating PI3K/Akt/mTOR signaling.

[Effects of electroacupuncture on N-methyl-D-aspartate receptor and mitogen activated protein kinase in spinal cord of rats with primary dysmenorrhea].

OBJECTIVE: To observe the effects of electroacupuncture (EA) on the expression levels of N-methyl-D-aspartate receptor (NMDAR), extracellular signal-regulated kinase (ERK)1/2, p38 mitogen activated protein kinase (p38 MAPK) and c-Jun N-terminal kinase (JNK) in the spinal cord of rats with primary dysmenoramia (PDM), so as to explore the underlying mechanism of EA treating PDM. METHODS: Thirty female SD rats were randomly divided into normal group, model group and EA group, with 10 rats in each group. The PDM rat model was established by subcutaneous injection of estradiol benzoate and oxytocin into the thigh. At the same time of modeling, rats in the EA group were treated with EA (50 Hz) at "Sanyinjiao" (SP36) and "Guanyuan" (CV4) once daily, 20 min each time, for 10 consecutive days. The writhing times, writhing score and writhing latency were observed within 30 min after oxytocin injection. The uterine pathological morphology was observed by HE staining, and pathological score was calculated. Serum prostaglandin F2α (PGF2α) and prostaglandin E2 (PGE2) were determined by ELISA. The protein expression levels of NMDAR, ERK1/2, p38MAPK and JNK in spinal cord were detected by Western blot. RESULTS: Compared with the normal group, the writhing times and writhing score were significantly increased (P<0.05); the endometrial epithelial cells showed vacuolar degeneration, death and hyperemia, the uterine pathological score was increased (P<0.05); the content of serum PGF2α and the ratio of PGF2α/PGE2 were significantly increased (P<0.01), while the content of serum PGE2 was significantly decreased (P<0.01); the expression levels of NMDAR, ERK1/2, p38MAPK and JNK in spinal cord were significantly increased (P<0.05, P<0.01) in the model group. Compared with the model group, the writhing times and writhing score were significantly decreased (P<0.05), the writhing latency was prolonged (P<0.05); the endometrial epithelial cells still showed vacuolar degeneration, death and hyperemia, and the uterine pathological score was decreased (P<0.01); the content of serum PGF2α and the ratio of PGF2α/PGE2 were significantly decreased (P<0.01), while the content of serum PGE2 was significantly increased (P<0.01); the protein expression levels of ERK1/2 and JNK in spinal cord were significantly decreased (P<0.01) in the EA group. CONCLUSION: EA intervention at SP36 and GV4 has obvious analgesic effect on PDM rats, and its mechanisms may be related to reducing serum prostaglandin, alleviating uterine inflammation, and inhibiting the protein expressions of NMDAR, ERK1/2, p38 MAPK and JNK in spinal cord.

Electroacupuncture treatment of primary dysmenorrhea: A randomized, participant-blinded, sham-controlled clinical trial protocol.

BACKGROUND: Primary dysmenorrhea in women is a common and serious public health problem with psychological and physical effects. Painkillers have adverse effects, such as tolerance, addiction, irritation of the digestive tract, and liver and kidney damage. Electroacupuncture has been used as alternative therapy, although with no (non-anecdotal) evidence of effectiveness. OBJECTIVE: This study aims to provide evidence for the feasibility and efficacy of electroacupuncture in the treatment of primary dysmenorrhea. Moreover, by observing changes in serum and urine metabolites, we will evaluate the putative mechanisms mediating electroacupuncture effects in primary dysmenorrhea. METHODS: This multicenter, randomized, participant-blinded, sham-controlled clinical trial including 336 women with primary dysmenorrhea is being conducted at three hospital centers in China and consists of a 12-week treatment and a 3-month follow-up. Women will undergo electroacupuncture (n = 168) or sham acupuncture (n = 168), beginning 7 days before their menstruation, once per day, until menstruation. Each menstrual cycle equals one course of treatment, and we will evaluate a total of three courses of treatment. The primary outcome of interest is the change in visual analogue scale scores before and after treatment. The secondary outcomes include changes in the numeric rating scale, Cox Menstrual Symptom Scale, traditional Chinese medicine symptoms, the Self-Rating Anxiety Scale, Self-Rating Depression Scale, and 36-Item Short Form questionnaire scores, and a safety evaluation. Moreover, we will preliminarily investigate the metabolomics mechanism as a potential mediator of the association between electroacupuncture and primary dysmenorrhea symptomology. DISCUSSION: We aim to find a suitable non-medicinal alternative for primary dysmenorrhea treatment to reduce reliance on non-steroidal anti-inflammatory drugs. TRIAL REGISTRATION: Chinese Clinical Trial Registry: ChiCTR2100054234; http://www.chictr.org.cn/.

[Effects of electroacupuncture on NLRP3 inflammasome and pyroptosis protein GSDMD in uterine tissue in rats with primary dysmenorrhea].

OBJECTIVE: To observe the effects of electroacupuncture (EA) on NLRP3 inflammasome and its downstream protein gastermin D (GSDMD) in rats with primary dysmenorrhea (PDM), and to explore the potential mechanism of EA on the treatment of PDM. METHODS: Forty healthy female SD rats without pregnancy were randomly divided into a control group, a model group, an EA group and an ibuprofen group, 10 rats in each group. PDM model was prepared by injection of estradiol benzoate and oxytocin. Except the control group, the rats in each group were subcutaneously injected with estradiol benzoate for 10 days, and oxytocin was injected on the 11th day. The rats in the EA group were intervened with EA (dense wave, frequency of 50 Hz) at "Guanyuan" (CV 4) and "Sanyinjiao" (SP 6) at the same time of modeling, once a day, 20 min each time, for 10 consecutive days. The rats in the ibuprofen group were treated with 0.8 mL of ibuprofen by gavage (concentration of ibuprofen solution was 1.25 mg/mL) for 10 consecutive days. After modeling, the writhing reaction was observed. After intervention, the HE staining method was used to observe the histological morphology of uterus and evaluate the pathological damage score of uterus; ELISA method was used to detect the serum levels of prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α); Western blot method was used to detect the protein expression of NLRP3, apoptosis related spot like protein (ASC), caspase-1, GSDMD, GSDMD-N and inflammatory factors (interleukin [IL]-1ß, IL-18) in uterine tissue. RESULTS: In the model group, a large number of vacuolar degeneration and death of endometrial epithelial cells, spiral arterioles congestion in lamina propria and neutrophil infiltration were observed. In the EA group, there was a small amount of vacuolar degeneration and death of endometrial epithelial cells, a small amount of spiral arterioles congestion in the lamina propria, and a small amount of neutrophils infiltration. In the ibuprofen group, there was very small number of degeneration and death of endometrial epithelial cells, and no obvious arterial congestion was found in lamina propria, and neutrophil infiltration was occasionally seen. Compared with the control group, in the model group the number of writhing was increased (P<0.01), the writhing reaction score and serum level of PGF2α and PGF2α/PGE2 value were increased (P<0.01), the level of PGE2 was decreased (P<0.01). Compared with the model group, in the EA group and the ibuprofen group the number of writhing were decreased (P<0.05), the latency of writhing was prolonged (P<0.01), the writhing reaction scores and serum levels of PGF2α and PGF2α/PGE2 values were decreased (P<0.05, P<0.01), the levels of PGE2 were increased (P<0.01). Compared with the control group, the protein expression of NLRP3, ASC, caspase-1, GSDMD, GSDMD-N, IL-1ß and IL-18 in the uterine tissues of rats was increased in the model group (P<0.01). Compared with the model group, the protein expression of NLRP3, ASC, caspase-1, GSDMD, GSDMD-N, IL-1ß and IL-18 in the uterine tissues of rats was decreased in the EA group and the ibuprofen group (P<0.01, P<0.05). There was no significant difference between the EA group and the ibuprofen group in the above indexes (P>0.05). CONCLUSION: EA could alleviate pain and uterine tissue injury in rats with PDM. The mechanism may be related to the inhibition of the activation of NLRP3 inflammasome in rat uterine tissues, thereby inhibiting pyroptosis and its inflammatory factors release.

[Effect of electroacupuncture intervention on relieving pain and inflammation by suppressing TLR4/NF-κB signaling in rats with primary dysmenorrhea].

OBJECTIVE: To investigate the mechanism of electroacupuncture(EA) intervention in rats with primary dysmenorrhea(PDM) based on the Toll-like receptor 4(TLR4)/nuclear factor(NF)-κB signaling pathway. METHODS: Forty female SD rats were randomly divided into blank control, model, EA and medication groups, with 10 rats in each group. PDM rat model was established by subcutaneous injection of estradiol benzoate combined with intraperitoneal injection of oxytocin. At the same time of model procedures, EA(50 Hz, dense wave) was applied to "Guanyuan" (CV4) and bilateral "Sanyinjiao" (SP6) of rats in the EA group, with needles retained for 20 min, for 10 consecutive days. Rats in the medication group received ibuprofen(125 mg/100 mL, 0.8 mL) by gavage for 10 consecutive days. At the 11th day, writhing behavior of rats was assessed. Uterine morphology was observed by eyes and uterine pathological changes were observed after HE staining. Content of prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) in serum and uterine tissues was detected by ELISA; NF-κB p65 positive expression in nucleus was detected by immunofluorescence; protein expression levels of TLR4, NF-κB p65, p-NF-κB p65 and inflammatory factors interleukin (IL) -1ß and IL-18 were detected by Western blot. RESULTS: After modeling, uterus tissues were congested and edematous, with necrosis of luminal epithelium, severe edema and extensive shedding of endometrium, nuclear pyknosis, fragmentation and disappearance, neutrophils infiltration, and slight expansion of glandular cavity, which was milder in the EA and the medication groups. Compared with the blank control group, writhing times, scores and incubation period, HE pathological scores, PGF2α contents in serum and uterine tissues, ratio of NF-κB p65 positive expression in nucleus, TLR4, NF-κB p65, p-NF-κB p65, IL-1ß and IL-18 protein expression levels in uterine tissues of rats in the model group were all significantly increased(P<0.01), while PGE2 contents in serum and uterine tissues were significantly decreased(P<0.01). Compared with the model group, writhing times and scores, HE pathological scores, PGF2α contents in serum and uterine tissues, ratio of NF-κB p65 positive expression in nucleus, TLR4, NF-κB p65, p-NF-κB p65, IL-1ß and IL-18 protein expression levels in uterine tissues of rats in the EA and medication group were all significantly decreased(P<0.01), while writhing incubation period, PGE2 contents in serum and uterine tissues were significantly increased(P<0.05, P<0.01). CONCLUSION: EA intervention could relieve inflammatory response and pain in PDM rats, which may be related to its effect in reducing TLR4 expression, inhibiting NF-κB activation and down-regulating inflammatory factors levels of IL-1ß and IL-18.

Effect of the annealing treatment on structural and transport properties of thermoelectric Smy(FexNi1-x)4Sb12thin films.

The crystallographic and transport properties of thin films fabricated by pulsed laser deposition and belonging to the Smy(FexNi1-x)4Sb12filled skutterudite system were studied with the aim to unveil the effect exerted by temperature and duration of thermal treatments on structural and thermoelectric features. The importance of annealing treatments in Ar atmosphere up to 523 K was recognized, and the thermal treatment performed at 473 K for 3 h was selected as the most effective in improving the material properties. With respect to the corresponding bulk compositions, a significant enhancement in phase purity, as well as an increase in electrical conductivity and a drop in room temperature thermal conductivity, were observed in annealed films. The low thermal conductivity, in particular, can be deemed as deriving from the reduced dimensionality and the consequent substrate/film interfacial stress, coupled with the nanometric grain size.

Multidimensional Pain Modulation by Acupuncture Analgesia: The Reward Effect of Acupuncture on Pain Relief.

Pain is an intrinsically unpleasant experience with features that protect an organism by promoting motivation and learning. Pain relief, a negative reinforcement of pain, is considered a reward and activates the brain's reward system. The reward circuit in the brain involves reward and pain. Acupuncture has a multidimensional and comprehensive regulating effect on chronic pain. However, the reward effect of acupuncture in relieving chronic pain and the mechanism of the brain reward circuit involved in acupuncture analgesia are not thoroughly studied. In this article, we have reviewed the definition of pain abnormalities and negative emotions in patients with chronic pain, the conceptual characteristics of analgesic reward, and the new progress in studying brain reward circuits and functions. Moreover, we have expounded on the critical clinical and scientific significance of studying the reward effect of acupuncture analgesia and related brain reward circuits, the pain mechanism obtained from human neuroimaging studies, and the survey results on the effects of acupuncture on reward/motivation circuits. Some viewpoints and suggestions on the reward effect of acupuncture analgesia and related reward circuits have been put forward to clarify the multidimensional characteristics and benign regulation of acupuncture analgesia. Studies on the reward effect of acupuncture in relieving chronic pain and the regulating effect of the brain reward loop on acupuncture analgesia help to deepen the clinical understanding of acupuncture analgesia, innovate the research concept of acupuncture analgesia, and provide help for further studies on the central mechanism of acupuncture in improving chronic pain in the future.

Moxibustion for Primary Dysmenorrhea: An Adjuvant Therapy for Pain Relief.

The latest spectrum of moxibustion disease shows that primary dysmenorrhea is a high-frequency symptom of moxibustion and that it is the dominant clinical disease. In the specific treatment methods, all types of moxibustion methods have been widely used, such as thermal, thunder fire, partitioned, and spreading moxibustion. Moxibustion plays a therapeutic role through its four mechanisms of action: heat, light, moxa smoke, and drug effects. The mechanism of moxibustion treatment for primary dysmenorrhea focuses on adjusting endocrine hormones, regulating immune function and neuro-related factors, and improving uterine microcirculation. In this study, based on the clinical evidence of different moxibustion methods for treating primary dysmenorrhea, the design model, intervention characteristics, and clinical outcomes were analyzed. Meanwhile, the brain effect mechanisms of different imaging methods were summarized from the perspective of neuroimaging. It was pointed out that the left anterior cingulate gyrus, left inferior parietal angular gyrus, and left superior gyrus may be the analgesic brain regions that regulate sensory, emotional, and cognitive aspects. Moreover, the neural circuits involved can be inferred: the frontal cortex-basal ganglia (the pea nucleus)-cerebral cortex, which mediates motivation and emotional drive, and the parietal lobe-basal ganglia-limbic lobe-frontal lobe, which is involved in neurotransmitter transport and emotional regulation and behavioral expression. There are still problems and deficiencies in studies on the mechanism of moxibustion treatment for primary dysmenorrhea. Studies should be strengthened on how moxibustion produces an effect. Attention should be paid to exploring how the spectrum range and peak in the light effect of moxibustion treat primary dysmenorrhea. Studies assessing the mechanisms of moxibustion treatment for primary dysmenorrhea should be conducted to provide an experimental basis and evidence-based medical evidence for clinical treatment.

Synthesis and Characterization of Al- and SnO2-Doped ZnO Thermoelectric Thin Films.

The effect of SnO2 addition (0, 1, 2, 4 wt.%) on thermoelectric properties of c-axis oriented Al-doped ZnO thin films (AZO) fabricated by pulsed laser deposition on silica and Al2O3 substrates was investigated. The best thermoelectric performance was obtained on the AZO + 2% SnO2 thin film grown on silica, with a power factor (PF) of 211.8 µW/m·K2 at 573 K and a room-temperature (300 K) thermal conductivity of 8.56 W/m·K. PF was of the same order of magnitude as the value reported for typical AZO bulk material at the same measurement conditions (340 µW/m·K2) while thermal conductivity κ was reduced about four times.

Investigation on the Power Factor of Skutterudite Smy(FexNi1-x)4Sb12 Thin Films: Effects of Deposition and Annealing Temperature.

Filled skutterudites are currently studied as promising thermoelectric materials due to their high power factor and low thermal conductivity. The latter property, in particular, can be enhanced by adding scattering centers, such as the ones deriving from low dimensionality and the presence of interfaces. This work reports on the synthesis and characterization of thin films belonging to the Smy(FexNi1-x)4Sb12-filled skutterudite system. Films were deposited under vacuum conditions by the pulsed laser deposition (PLD) method on fused silica substrates, and the deposition temperature was varied. The effect of the annealing process was studied by subjecting a set of films to a thermal treatment for 1 h at 423 K. Electrical conductivity σ and Seebeck coefficient S were acquired by the four-probe method using a ZEM-3 apparatus performing cycles in the 348-523 K temperature range, recording both heating and cooling processes. Films deposited at room temperature required three cycles up to 523 K before being stabilized, thus revealing the importance of a proper annealing process in order to obtain reliable physical data. XRD analyses confirm the previous result, as only annealed films present a highly crystalline skutterudite not accompanied by extra phases. The power factor of annealed films is shown to be lower than in the corresponding bulk samples due to the lower Seebeck coefficients occurring in films. Room temperature thermal conductivity, on the contrary, shows values comparable to the ones of doubly doped bulk samples, thus highlighting the positive effect of interfaces on the introduction of scattering centers, and therefore on the reduction of thermal conductivity.

Abnormal Topological Network in Parkinson's Disease With Impulse Control Disorders: A Resting-State Functional Magnetic Resonance Imaging Study.

In recent years, neuroimaging evidence shows that the brains of Parkinson disease (PD) with impulse control disorders (ICDs) patients have functional disconnection changes. However, so far, it is still unclear whether the topological organization is damaged in PD patients with ICD. In this study, we aimed to explore the functional brain network in 18 patients with PD with ICDs (PD-ICD) and 18 patients with PD without ICDs (PD-nICD) by using functional magnetic resonance imaging and graph theory approach. We found that the PD-ICD patients had increased clustering coefficient and characteristic path length, while decreased small-world index compared with PD-nICD patients. Furthermore, we explored the hypothesis whether the abnormality of the small-world network parameters of PD-ICD patients is accompanied by the change of nodal centrality. As we hypothesized, the nodal centralities of the default mode network, control network, and dorsal attention network were found to be significantly damaged in the PD-ICD group compared with the PD-nICD group. Our study provides more evidence for PD-ICD patients' brain network abnormalities from the perspective of information exchange, which may be the underlying pathophysiological basis of brain abnormalities in PD-ICD patients.

Lactoferrin may inhibit the development of cancer via its immunostimulatory and immunomodulatory activities (Review).

Lactoferrin (Lf) is secreted by ectodermal tissue and has a structure similar to that of transferrin. Although Lf seems to be multifunctional, its main function is related to the natural defense system of mammals. The present review aims to highlight the major actions of Lf, including the regulation of cell growth, the inhibition of toxic compound formation, the removal of harmful free radicals and its important role in immune response regulation. Moreover, Lf has antibacterial, antiviral, antioxidant, anticancer and anti­inflammatory activities. In addition, the use of Lf for functionalization of drug nanocarriers, with emphasis on tumor­targeted drug delivery, is illustrated. Such effects serve as an important theoretical basis for its future development and application. In neurodegenerative diseases and the brains of elderly people, Lf expression is markedly upregulated. Lf may exert an anti­inflammatory effect by inhibiting the formation of hydroxyl free radicals. Through its antioxidant properties, Lf can prevent DNA damage, thereby preventing tumor formation in the central nervous system. In addition, Lf specifically activates the p53 tumor suppressor gene.

Characterization of ferroptosis signature to evaluate the predict prognosis and immunotherapy in glioblastoma.

BACKGROUND: Glioblastoma (GBM) is the most common type of brain cancer with poor survival outcomes and unsatisfactory response to current therapeutic strategies. Recent studies have demonstrated that ferroptosis-related genes (FRGs) are linked with the occurrence and development of GBM and may become promising biological indicators in GBM therapy. METHODS: We systematically assessed the relationship between FRGs expression profiles and prognosis in glioma patients based on the Cancer Genome Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA) datasets to establish a risk score model according to the gene signature of multiple survival-associated DEGs. Further, the differences between the tumor microenvironment score, immune cell infiltration, immune checkpoint expression levels, and drug sensitivity in the high- and low-risk group are analyzed through a variety of algorithms in R software. RESULTS: GBM patients were divided into two subgroups (high- and low-risk) according to the established risk score model. Patients in the high-risk group showed significantly reduced overall survival compared with those in the low-risk group. Also, we found that the high-risk group showed higher ImmuneScore and StromalScore, while different subgroups have significant differences in immune cell infiltration, immune checkpoint expression levels, and drug sensitivity. In summary, we developed and validated an FRGs risk model, which served as an independent prognostic indicator for GBM. Besides, the two subgroups divided by the model have significant differences, which provides novel insights for further studies as well as the personalized treatment of patients.

Novel Biomarker Genes for Prognosis of Survival and Treatment of Glioma.

Glioblastoma multiforme (GBM) is the most aggressive malignant primary central nervous system tumor. Although surgery, radiotherapy, and chemotherapy treatments are available, the 5-year survival rate of GBM is only 5.8%. Therefore, it is imperative to find novel biomarker for the prognosis and treatment of GBM. In this study, a total of 141 differentially expressed genes (DEGs) in GBM were identified by analyzing the GSE12657, GSE90886, and GSE90598 datasets. After reducing the data dimensionality, Kaplan-Meier survival analysis indicated that expression of PTPRN and RIM-BP2 were downregulated in GBM tissues when compared with that of normal tissues and that the expression of these genes was a good prognostic biomarker for GBM (p<0.05). Then, the GSE46531 dataset and the Genomics of Drug Sensitivity in Cancer (GDSC) database were used to examine the relationship between sensitivity radiotherapy (RT) and chemotherapy for GBM and expression of PTPRN and RIM-BP2. The expression of PTPRN was significantly high in RT-resistant patients (p<0.05) but it was not related to temozolomide (TMZ) resistance. The expression level of RIM-BP2 was not associated with RT or TMZ treatment. Among the chemotherapeutic drugs, cisplatin and erlotinib had a significantly good treatment effect for glioma with expression of PTPRN or RIM-BP2 and in lower-grade glioma (LGG) with IDH mutation. (p < 0.05). The tumor mutational burden (TMB) score in the low PTPRN expression group was significantly higher than that in the high PTPRN expression group (p=0.013), with a large degree of tumor immune cell infiltration. In conclusion, these findings contributed to the discovery process of potential biomarkers and therapeutic targets for glioma patients.

LncRNA GAS5 regulates epithelial-mesenchymal transition and viability of glioma cells by targeting microRNA-106b and regulating PTEN expression.

LncRNA growth arrest special 5 (GAS5) and microRNA-106b (miR-106b) have been reported to be involved in the regulation of gliomas. However, their precise mechanisms in regulating the progression and development of gliomas remain unclear. We aimed to investigate the interaction between GAS5 and miR-106b, and their influence on the proliferation, migration, and invasion of gliomas cells. Western blotting and qRT-PCR were applied for measuring expression of protein and mRNA, respectively. The proliferation, migration, and invasion of cells were measured by MTT, wound healing, and transwell assays, respectively. Dual luciferase reporter assay was applied for confirming the binding site between miR-106b and GAS5, miR-106b and PTEN. Significant higher expression of miR-106b, and lower expression of GAS5 and PTEN in the glioma tissues were observed. The binding sites between GAS5 and miR-106b, miR-106b and PTEN were identified. GAS5 could regulate the expression of PTEN through targeting miR-106b, and further influence EMT process, and the proliferation, migration, and invasion of gliomas cells. Meanwhile, PTEN could remarkably inhibited the proliferation, migration and invasion of glioma cells. The influence of PTEN on glioma cells and EMT was similar to GAS5. GAS5 could regulate the EMT process, and the migration of gliomas cells through miR-106b targeting PTEN. Therefore, our findings may provide a new thought for the study of pathogenesis and treatment of glioma.

Metabolite and Transcriptome Profiling on Xanthine Alkaloids-Fed Tea Plant (Camellia sinensis) Shoot Tips and Roots Reveal the Complex Metabolic Network for Caffeine Biosynthesis and Degradation.

While caffeine is one of the most important bioactive metabolites for tea as the most consumed non-alcohol beverage, its biosynthesis and catabolism in tea plants are still not fully understood. Here, we integrated purine alkaloid profiling and transcriptome analysis on shoot tips and roots fed with caffeine, theophylline, or theobromine to gain further understanding of caffeine biosynthesis and degradation. Shoot tips and roots easily took up and accumulated high concentrations of alkaloids, but roots showed much faster caffeine and theophylline degradation rates than shoot tips, which only degraded theophylline significantly but almost did not degrade caffeine. Clearly feedback inhibition on caffeine synthesis or inter-conversion between caffeine, theophylline, and theobromine, and 3-methylxanthine had been observed in alkaloids-fed shoot tips and roots, and these were also evidenced by significant repression of TCS and MXMT genes critical for caffeine biosynthesis. Among these responsively repressed genes, two highly expressed genes TCS-4 and TCS-8 were characterized for their enzyme activity. While we failed to detect TCS-4 activity, TCS-8 displayed N-methyltransferase activities towards multiple substrates, supporting the complex metabolic network in caffeine biosynthesis in tea plants since at least 13 TCS-like N-methyltransferase genes may function redundantly. This study provides new insight into complex metabolic networks of purine alkaloids in tea plants.

Guanine deaminase provides evidence of the increased caffeine content during the piling process of pu'erh tea.

Wet piling is a key process for producing pu'erh tea because various components change under the action of microorganisms. Among these components, caffeine content is increased. Evidence has indicated a salvage pathway for caffeine biosynthesis in microbes, in which xanthine is methylated in the order of N-3 → N-1 → N-7. In addition, guanine can be used to synthesize xanthine through guanine deaminase (EC: 3.5.4.3). In this study, we investigated the variation in caffeine content during piling fermentation with supplementary guanine, 15N-labeled guanine and xanthine. We cloned the guanine deaminase gene (GUD1) from Saccharomyces cerevisiae (one dominant strain in piling fermentation). The results revealed that [15N]xanthine could be synthesized from [15N]guanine, and [15N]caffeine was also detected during piling with supplementary [15N]xanthine. Furthermore, ScGUD1 could catalyze the conversion of guanine to xanthine, which is likely to be methylated for caffeine synthesis under microorganism action. The obtained results revealed the mechanism underlying the increased caffeine content during piling of pu'erh tea.

[Investigation on the convertion between ginkgolide B and its derivative].

A derivative decomposed from ginkgolide B (GB) was isolated by semi-preparative chromatography. The results of the high performance liquid chromatography (HPLC) showed that the retention time of the derivative was approximately three times of that of GB with ultraviolet detection (UV), and the derivative can not exist independently without the presence of GB. The UV spectrum showed that the lamdamax of the derivative was 212.1 nm and the epsilonmax was 2.29 x 10(4), which were approximately 100 times higher than those of GB. This means a new conjugation bond has been formed and that means that the conjugation bond's pii-pi* electron jump occured in the derivative. Liquid chromatography-mass spectrometry showed the molecular ion peaks of the derivative in the positive mode and negative mode were m/z 429.1 (M + Na)+ and m/z 405.2 (M-H)-, respectively. The relative molecular mass of the derivative is 406.2. The derivative showed the same mode of fragment ion as GB in the mass spectrum, which might be due to the loss of a H2O from GB. Thermal stability of GB was greater than that of the derivative. They were both easily to be dissolved in dilute alkali. When the pH gradually increases, the ring-opening speed of the derivative is higher than that of GB. The derivative was obviously affected by solvent and higher temperatures. When GB was dissolved in PEG, the peak of the derivative disappeared at 50 degrees C for 15 h or at 120 degrees C for 4 h. Besides the main peak of GB, there appeared a small peak with a retention time of 1.2-3.0 min after heating 4 h at 120 degrees C. The result showed that the derivative was in a high energy state, and GB had a better stabilization. They coexisted and converted to each other. Under some specific conditions, the derivative could all convert to GB.