Representation and control of pain and itch by distinct prefrontal neural ensembles.

Pain and itch are two closely related but essentially distinct sensations that elicit different behavioral responses. However, it remains mysterious how pain and itch information is encoded in the brain to produce differential perceptions. Here, we report that nociceptive and pruriceptive signals are separately represented and processed by distinct neural ensembles in the prelimbic (PL) subdivision of the medial prefrontal cortex (mPFC) in mice. Pain- and itch-responsive cortical neural ensembles were found to significantly differ in electrophysiological properties, input-output connectivity profiles, and activity patterns to nociceptive or pruriceptive stimuli. Moreover, these two groups of cortical neural ensembles oppositely modulate pain- or itch-related sensory and emotional behaviors through their preferential projections to specific downstream regions such as the mediodorsal thalamus (MD) and basolateral amygdala (BLA). These findings uncover separate representations of pain and itch by distinct prefrontal neural ensembles and provide a new framework for understanding somatosensory information processing in the brain.

[Eye acupuncture improves cerebral ischemia reperfusion injury by improving autophagy via ATF6 pathway in cerebral ischemia reperfusion injury rats].

OBJECTIVE: To explore the effect of eye acupuncture on autophagy and expressions of autophagy-related proteins Beclin1, LC3B, ATF6 and XBP1 in the infarction area of brain tissue in rats with acute cerebral ischemia-reperfusion injury (CIRI), so as to explore its mechanisms underlying improvement of CIRI. METHODS: Male SD rats were randomly divided into sham operation, model and eye acupuncture groups (n=16 in each group). The CIRI model was prepared by occlusion of the middle cerebral artery. Eye acupuncture stimulation was applied to bilateral "Gan"(Liver), "Shangjiao"(Upper-energizer), "Xiajiao"(Lower-energizer) and "Shen"(Kidney) regions at 0, 12 and 24 h after CIRI, 30 min each time. The neurological deficit score was given by referring to Longa's method, and TTC staining used to determine the success of model replication. After the treatment, the pathological changes of the cerebral infarction area were observed under light microscope, and the autophagosomes were observed by electron microscope. The protein expression levels of LC3B, Beclin1, ATF6 and XBP1 in the infarction area of brain tissue were detected by Western blot. The immunoactivity of Beclin1 and the immunofluorescence density of ATF6 and XBP1 in the infarction area of brain tissue were determined by immunohistochemistry. RESULTS: The Longa's score, and the protein expression levels of LC3B, Beclin1, ATF6 and XBP1 and immunoactivity or immunofluorescence density of Beclin1, ATF6 and XBP1 were significantly higher in the model group than those in the sham operation group (P<0.01), and considerably lower in the eye acupuncture group than those in the model group (P<0.01). Under light microscope, the model group had typical ethmoidal reticular cerebral infarction, while the eye acupuncture group had significantly smaller areas and clearer edges. Under electron microscope, there were more autophagosomes in the cytoplasm of neurons in the model group, and fewer autophagosomes in the eye acupuncture group (in contrast to the model group). CONCLUSION: Eye acupuncture can improve the neurological function and mitigate cerebral injury in CIRI rats which may be associated with its function in inhibiting autophagy in the brain tissue by regulating ATF6 pathway.

Multi-omic analysis in transgenic mice implicates omega-6/omega-3 fatty acid imbalance as a risk factor for chronic disease.

An unbalanced increase in dietary omega-6 (n-6) polyunsaturated fatty acids (PUFA) and decrease in omega-3 (n-3) PUFA in the Western diet coincides with the global rise in chronic diseases. Whether n-6 and n-3 PUFA oppositely contribute to the development of chronic disease remains controversial. By using transgenic mice capable of synthesizing PUFA to eliminate confounding factors of diet, we show here that alteration of the tissue n-6/n-3 PUFA ratio leads to correlated changes in the gut microbiome and fecal and serum metabolites. Transgenic mice able to overproduce n-6 PUFA and achieve a high tissue n-6/n-3 PUFA ratio exhibit an increased risk for metabolic diseases and cancer, whereas mice able to convert n-6 to n-3 PUFA, and that have a lower n-6/n-3 ratio, show healthy phenotypes. Our study demonstrates that n-6 PUFA may be harmful in excess and suggests the importance of a low tissue n-6/n-3 ratio in reducing the risk for chronic diseases.

[Effect of Biochar on Root Morphological Characteristics of Wetland Plants and Purification Capacity of Constructed Wetland].

A constructed wetland with Acorus calamus L. was built. Straw biochar, reed biochar, and sawdust biochar was added into the constructed wetland individually to study the effect of different biochars on the root morphology, dissolved oxygen, and purification ability of the constructed wetland. The results show that the total root length, total projection area, total volume, total surface area, root number, branch number, and root dry weight of Acorus calamus L. significantly increased when all three kinds of biochar were added into the constructed wetland (P<0.05). Similarly, adding the biochars into the constructed wetland also significantly increased dissolved oxygen content in the wetland (P<0.05). Addition of sawdust biochar into the constructed wetland increased the root length, projection area, surface area, total volume, number of root tips, number of branches, and root dry weight of Acorus calamus L. by 96.1%, 106.2%, 185.6%, 172.5%, 75.3%, 121.6%, and 84.9%, respectively. After adding biochars into the constructed wetland, the root morphology of Acorus calamus L. and dissolved oxygen content was significantly correlated with removal rate of TN, TP, and COD, respectively. Addition of sawdust biochar into the constructed wetland significantly increased the removal rates of total nitrogen, total phosphorus, and COD when the hydraulic load was 0.022 m3·(m2·d)-1 (P<0.05). These results suggested that the addition of sawdust biochar to the constructed wetland increased the root growth of Acorus calamus L. and enhanced dissolved oxygen content, resulting in purification capacity of the constructed wetland.

Structure Identification and In Vitro Anticancer Activity of Lathyrol-3-phenylacetate-5,15-diacetate.

Natural products from the genus Euphorbia show attention-attracting activities, such as anticancer activity. In this article, classical isolation and structure identification were used in a study on Caper Euphorbia Seed. Subsequently, MTT and wound healing assays, flow cytometry, western blotting, Hoechst 33258 staining and fluorescence microscopy examination were applied to investigate the anticancer activity of the obtained compounds. In a result, lathyrol-3-phenyl- acetate-5,15-diacetate (deoxy Euphorbia factor L1, DEFL1) was isolated from Caper Euphorbia Seed. Moreover, the NMR signals were totally assigned. DEFL1 showed potent inhibition against lung cancer A549 cells, with an IC50 value of 17.51 ± 0.85 µM. Furthermore, DEFL1 suppressed wound healing of A549 cells in a concentration-dependent manner. Mechanically, DEFL1 induced apoptosis, with involvement of an increase of reactive oxygen species (ROS), decrease of mitochondrial membrane potential (ΔΨm), release of cytochrome c, activity raise of caspase-9 and 3. Characteristic features of apoptosis were observed by fluorescence microscopy. In summary, DEFL1 inhibited growth and induced apoptosis in lung cancer A549 cells via a mitochondrial pathway.

[Effect of eye-acupuncture on cerebral intracellular adhesion molecule-1 expression in rats with acute cerebral ischemia-reperfusion injury].

OBJECTIVE: To investigate the effect of the eye-acupuncture therapy on cerebral intercellular adhesion molecule-1 (ICAM-1) expression in rats with acute cerebral ischemia-reperfusion injury (CI/RI) so as to reveal its underlying mechanism. METHODS: Forty male SD rats were randomly and equally divided into normal, sham operation (sham), model and eye-acupuncture groups. Acute CI/RI model was established by occlusion of the middle cerebral artery with a modified suture-blocking method. Filiform needles were inserted into the "Qan" (Liver Area), "Shangjiao" (Upper Energizer Area), "Xiajiao" (Lower Energizer Area), etc. of the eye-acupuncture acupoints, and retained for 20 min. The treatment was conducted twice daily, 7 times altogether. Seventy-two hours after Acute CI/RI, the neurological behavior was assessed by ZeaLonga neurophysical impairment scale. Cerebral ICAM-1 protein and ICAM-1 mRNA expression levels were detected by Western blot (Penumbra area of the right cerebral infarction region) and real time-polymerase chain reaction (PCR) methods, respectively. RESULTS: After modeling, the neurophysical impairment score of the model group was increased from 0 to (2.63 +/- 0.92) points on the first day and (2.63 +/- 0.74) points 72 h following Acute CI/RI. Correspondingly, cerebral ICAM-1 protein and mRNA expression was up-regulated considerably in the model group than in the normal control group (P < 0. 01). In comparison with the model group, the neurologic impairment score, and cerebral ICAM-1 protein and mRNA expression levels of the eye-acupuncture group were down-regulated obviously (P < 0.01, P < 0.05). No significant differences were found between the normal control and sham groups in neurologic impairment score and the expression levels of the ICAM-1 protein and ICAM-1 mRNA(P > 0.05). CONCLUSION: Eye-acupuncture therapy can improve the cerebral ischemia-reperfusion injury induced behavior changes, which may be related to its effects in down-regulating the expression of cerebral ICAM-1 protein and gene.

A new method for the treatment of chromite ore processing residues.

Chromite ore processing residue (COPR), derived from the so-called high-lime processing of chromite ore, contains a significant fraction of a leachable Cr(VI) which is harmful to human being and other organisms. In recent years, the concern over environmental pollution from the waste residue containing Cr(VI) has become a major problem for the chromium chemical industry. The main purpose of this investigation is to evaluate a new method for remediation of Cr(VI) in COPR. COPR was mixed with reductants, sucrose, starch or flour, and was calcinated at elevated temperatures in inertial gas. Effects of temperature, dosages of reductants and time on Cr(VI) reduction were investigated. Above 500 degrees C, Cr(VI) can be completely reduced to Cr(III).

A novel fusion suicide gene yeast CDglyTK plays a role in radio-gene therapy of nasopharyngeal carcinoma.

To investigate a novel suicide gene for nasopharyngeal carcinoma (NPC) therapy, the yCDglyTK gene was constructed by fusing yeast cytosine deaminase (CD) and herpes simplex type 1 thymidine kinase. The expression of the yCDglyTK gene was detected by RT-PCR and Western blotting, and its bioactivity was demonstrated by an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. An animal study was carried out in which BALB/C nude mice bearing yCDglyTK gene-modified tumors were treated with prodrugs and radiation. Our results revealed that the yCDglyTK gene could be expressed in CNE-2 cells in vitro. In MTT analysis, at the transfection rate of 10%, 66% cells were killed. The synergistic effect of CD and TK showed 91% of yCDglyTK-transfected cells were killed with the treatment of 5-fluorocytosine (5-FC) alone, 60% killed with ganciclovir (GCV) alone, and 75% killed with 5-FC and GCV together. In vivo, the tumor volume in all of the four prodrugs and/or radiation-treated groups were significantly different from that in the PBS-controlled group (P<.01); also yCDglyTK+prodrug+radiation group was different from the other three groups (P<.05). Our findings suggested there was a synergistic antitumor effect when combining suicide gene therapy and radiation, and yCDglyTK has potent antitumor efficacy and may be a candidate suicide gene for cancer therapy.

Three novel mutations of the spastin gene in Chinese patients with hereditary spastic paraplegia.

BACKGROUND: Hereditary spastic paraplegia is a group of genetically heterogeneous neurodegenerative disorders characterized by progressive spasticity of the lower limbs. The most common form of hereditary spastic paraplegia is caused by mutations in the spastin gene (SPG4), which encodes spastin, an adenosine triphosphatase associated with various cellular activities protein. OBJECTIVE: To investigate the Chinese patients with hereditary spastic paraplegia for mutations in SPG4. METHODS: DNA samples from 31 unrelated patients were analyzed for mutations in SPG4 by single-strand conformation polymorphism analysis and direct sequencing. All DNA samples were screened for mutations by the polymerase chain reaction, followed by electrophoresis and silver staining. Each new variant identified was analyzed in 50 control subjects to determine whether it is a polymorphism or a mutation. RESULTS: Three novel mutations were detected in 4 affected individuals, including 2 missense mutations (T1258A and A1293G) and 1 deletion mutation (1668-1670delCTA). CONCLUSIONS: To our knowledge, this is the first report of SPG4 mutations in the People's Republic of China. The percentage of involved Chinese families with autosomal dominant hereditary spastic paraplegia with an SPG4 mutation is 18% (4/22), lower than the estimated 40% linked to this locus.

[Expression of cyclooxygenase-2 mRNA and identification of its spliceriant in human myometrium obtained from women in labor].

OBJECTIVE: To investigate the expression of cyclooxygenase-2 (COX-2) in human lower segments of myometrium obtained from women in labor and those not in labor and identify the splice variant of COX-2. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) was used for investigating the expression of COX-2. According to the sequence of rat COX-2 splice variant, the primers were designed and synthesized, then the splice variant of COX-2 in human myometrium from woman in labor was identified, cloned into vector and sequenced. RESULTS: The results showed that (1) The Expression of COX-2 mRNA was lower in human myometrium obtained from women who were not in labor than those in labor. (2) A new band of COX-2 was obtained in myometrium from a woman in labor. The fragment includes an unspliced intron, which locates between exons 7 and 8. CONCLUSION: COX-2 gene is not only expressed highly in human myometrium from women in labor, but also produced splicing variant by alternative splicing.