Hyperatins A-D, highly oxidized polycyclic polyprenylated acylphloroglucinols from Hypericum perforatum L. with hypoglycemic potential in liver cells.

Hyperatins A-D (1-4), four previously undescribed polycyclic polyprenylated acylphloroglucinols, were isolated from Hypericum perforatum L. (St. John's wort). Compound 1 possessed a unique octahydroindeno[1,7a-b]oxirene ring system with a rare 2,7-dioxabicyclo[2.2.1]heptane fragment. Compounds 2-4 had an uncommon decahydrospiro[furan-3,7'-indeno[7,1-bc]furan] ring system. Their structures were established by spectroscopic analyses and X-ray crystallography. Plausible biosynthetic pathways of 1-4 were also proposed. Compounds 1 and 2 exerted promising hypoglycemic activity by inhibiting glycogen synthase kinase 3 expression in liver cells.

Discovery of bioactive polycyclic polyprenylated acylphloroglucinols with adamantine/homoadamantane skeletons from Hypericum wilsonii.

In this work, nine previous undescribed polycyclic polyprenylated acylphloroglucinols with adamantine/homoadamantane skeletons, cumilcinols A-I (1-9), along with six known analogues, were isolated and identified from the stems, leaves and flowers of Hypericum wilsonii. Their structures were determined by HRESIMS, NMR spectroscopic analysis, single-crystal X-ray crystallography as well as electronic circular dichroism calculations and comparisons. Compound 2 formed a unique furan ring bearing a rare acetal functionality. In bioassays, hyperacmosin G (13) could significantly inhibit the production of NO in LPS-stimulated RAW264.7 cell (IC50 = 4.350 ± 1.146 µM), and increased expression of related transcription factors at the gene level, inhibit the nuclear translocation of NF-κBp65, and reduce the protein expression of COX-2. Additionally, compound 5 showed significant inhibitory activity on Con A-induced T-lymphocyte proliferation (IC50 = 4.803 ± 3.149 µM), and treatment of 5 could reduce the increased ratio of CD4 and CD8 subpopulations induced by Con A in vitro. Those results indicated 13 possesses potential anti-inflammatory activity, and 5 exhibits a certain degree of immunosuppressive activity.

Influencing Factors of Recurrence of Lumbar Disc Herniation After Percutaneous Transforaminal Endoscopic Discectomy: A Meta-Analysis.

Background: Lumbar disc herniation (LDH) remains one of the extremely common diseases in the elderly population, and despite the fact that percutaneous transforaminal endoscopic discectomy (PTED) can be an effective treatment for LDH, prognostic recurrence of the patients is still a clinical problem that needs to be addressed. Objective: To perform a meta-analysis of the influencing factors of disease recurrence after PTED for LDH to provide evidence for clinical practice. Methods: By screening the PubMed, EMbase, and Cochrane Library databases for relevant studies on disease recurrence after PTED for LDH, we extracted the authors, publication time, outcome measures, and other indicators were extracted for meta-analyses using RevMan 5.3 software. Results: The online retrieval and rigorous screening returned 8 valid articles for analysis, all with high reference value, as their Newcastle Ottawa Scale (NOS) scores were above 6. According to meta-analyses, there were no differences in gender and LDH type and location among patients with LDH recurrence after PTED treatment (P > .05); however, statistical significance was present in Pfirrmann grading, incomplete nucleus pulposus removal during surgery, and Modic changes (P < .05), indicating that these indexes were the influencing factors of LDH recurrence. Conclusions: Pfirrmann grading, incomplete nucleus pulposus removal during surgery, and Modic changes are related factors affecting LDH recurrence after PTED.

[Alcohol extract of root and root bark of Toddalia asiatica alleviates CIA in rats through anti-inflammatory and proapoptotic effects].

This study aims to investigate the therapeutic effect of alcohol extract of root and root bark of Toddalia asiatica(TAAE) on collagen-induced arthritis(CIA) in rats through phosphatidylinoinosidine-3 kinase/protein kinase B(PI3K/Akt) signaling pathway. To be specific, CIA was induced in rats, and then the rats were treated(oral, daily) with TAAE and Tripterygium Glycoside Tablets(TGT), respectively. The swelling degree of the hind leg joints was scored weekly. After 35 days of administration, the histopathological changes were observed based on hematoxylin and eosin(HE) staining. Enzyme-linked immunosorbent assay(ELISA) was employed to detect the levels of cytokines [tumor necrosis factor-α(TNF-α), interleukin(IL)-6)]. Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL) staining was performed to detect the apoptosis of synoviocytes in rats. Western blot was used to detect the expression levels of apoptosis-related proteins B-cell lymphoma 2(Bcl-2)-associated X(Bax), Bcl-2, and caspase-3 and pathway-related proteins phosphoinositide 3-kinase(PI3K), phosphorylated(p)-PI3K, protein kinase B(Akt), and p-Akt. RT-qPCR was conducted to examine the mRNA levels of Bax, Bcl-2, caspase-3, TNF-α, IL-6, and IL-1ß and pathway-related proteins PI3K, p-PI3K, Akt, and p-Akt. TAAE can alleviate the joint swelling in CIA rats, reduce serum levels of inflammatory cytokines, improve synovial histopathological changes, promote apoptosis of synoviocytes, and inhibit synovial inflammation. In addition, RT-qPCR and Western blot results showed that TAAE up-regulated the level of Bax, down-regulated the level of Bcl-2, and activated caspase-3 to promote apoptosis in synoviocytes. TAAE effectively down-regulated the protein levels of p-PI3K and p-Akt. In this study, TAAE shows therapeutic effect on CIA in rats and reduces the inflammation. The mechanism is that it suppresses PI3K/Akt signaling pathway and promotes synoviocyte apoptosis. Overall, this study provides a new clue for the research on the anti-inflammatory mechanism of TAAE and lays a theoretical basis for the better clinical application of TAAE in the treatment of inflammatory and autoimmune diseases.

Isolation of a degrading strain of Fusarium verticillioides and bioremediation of glyphosate residue.

Glyphosate is a broad-spectrum and nonselective organophosphorus herbicide that inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), an enzyme in the shikimate pathway in plants. A glyphosate-resistant fungus identified as Fusarium verticillioides was screened from soil subjected to long-term glyphosate application, and this fungus could grow in inorganic salt medium containing 90 mmol/L glyphosate. The optimum culture conditions identified via the response surface curve method were 28 °C and pH 7.0. The target gene epsps was cloned in this study, and the open reading frame contained 1170 nucleotides and putatively encoded 389 amino acid residues. Phylogenetic analysis showed that this gene belonged to class I, genes naturally sensitive to glyphosate. q-PCR confirmed that the relative expression level of the epsps gene was low, and no significant difference in expression was observed among different glyphosate concentrations at 12 h or 48 h. On day 28, the degradation by Fusarium verticillioides C-2 of sterilized soil and unsterilized soil supplemented with 60 mg/kg glyphosate reached 72.17% and 89.07%, respectively, and a significant difference was observed between the treatments with and without the glyphosate-degrading strain. The recovery of soil dehydrogenase activity after the addition of Fusarium verticillioides was significantly higher than that in the absence of the degrading fungus on the 28th day. The results showed that C-2 is a highly effective glyphosate-degrading strain with bioremediation potential for glyphosate-contaminated soil.

Grey matter connectome abnormalities and age-related effects in antipsychotic-naive schizophrenia.

BACKGROUND: Convergent evidence is increasing to indicate progressive brain abnormalities in schizophrenia. Knowing the brain network features over the illness course in schizophrenia, independent of effects of antipsychotic medications, would extend our sight on this question. METHODS: We recruited 237 antipsychotic-naive patients with schizophrenia range from 16 to 73 years old, and 254 healthy controls. High-resolution T1 weighted images were obtained with a 3.0T MR scanner. Grey matter networks were constructed individually based on the similarities of regional grey matter measurements. Network metrics were compared between patient groups and healthy controls, and regression analyses with age were conducted to determine potential differential rate of age-related changes between them. FINDINGS: Nodal centrality abnormalities were observed in patients with untreated schizophrenia, particularly in the central executive, default mode and salience networks. Accelerated age-related declines and illness duration-related declines were observed in global assortativity, and in nodal metrics of left superior temporal pole in schizophrenia patients. Although no significant intergroup differences in age-related regression were observed, the pattern of network metric alternation of left thalamus indicated higher nodal properties in early course patients, which decreased in long-term ill patients. INTERPRETATIONS: Global and nodal alterations in the grey matter connectome related to age and duration of illness in antipsychotic-naive patients, indicating potentially progressive network organizations mainly involving temporal regions and thalamus in schizophrenia independent from medication effects. FUNDING: The National Natural Science Foundation of China, Sichuan Science and Technology Program, the Fundamental Research Funds for the Central Universities, Post-Doctor Research Project, West China Hospital, Sichuan University , the Science and Technology Project of the Health Planning Committee of Sichuan, Postdoctoral Interdisciplinary Research Project of Sichuan University and 1.3.5 Project for Disciplines of Excellence, West China Hospital, Sichuan University.

Molecular mechanism of Epicedium treatment for depression based on network pharmacology and molecular docking technology.

BACKGROUND: Increasing attention has been paid to the effect of Epimedium on the nervous system, particularly anti-depression function. In the present study, we applied network pharmacology to introduce a testable hypothesis on the multi-target mechanisms of Epicedium against depression. METHODS: By reconstructing the network of protein-protein interaction and drug-component-target, we predicted the key protein targets of Epicedium for the treatment of depression. Then, through molecular docking, the interaction of the main active components of Epicedium and predicted candidate targets were verified. RESULTS: Nineteen active compounds were selected from Epicedium. There were 200 targets associated with Epicedium and 537 targets related to depression. The key targets of Epicedium for treating depression were IL6, VEGFA, AKT1, and EGF. According to gene ontology functional enrichment analysis, 22 items of biological process (BP), 13 items of cell composition (CC) and 9 items of molecular function (MF) were obtained. A total of 56 signaling pathways (P < 0.05) were identified by Kyoto Encyclopedia of Genes and Genomes analysis, mainly involving depression-related pathways such as dopaminergic synapse, TNF signaling pathway, and prolactin signaling pathway. The results of molecular docking showed that the most important activity components, including luteoklin, quercetin and kaempferol, were well combined with the key targets. CONCLUSIONS: Luteoklin, quercetin, kaempferol and other active compounds in Epicedium can regulate multiple signaling pathways and targets such as IL6, AKT1, and EGF, therefore playing therapeutic roles in depression.

The mechanisms of Chuanxiong Rhizoma in treating spinal cord injury based on network pharmacology and experimental verification.

BACKGROUND: Chuanxiong Rhizoma (CR) is a common traditional Chinese medicine (TCM) that has been widely used in the treatment of spinal cord injury (SCI). However, the underlying molecular mechanism of CR is still largely unknown. This study was designed to explore the bioactive components and the mechanism of CR in treating SCI based on a network pharmacology approach and experimental validation. METHODS: First, the active compounds and related target genes in CR were screened from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. Subsequently, the corresponding target genes of SCI were collected by the Therapeutic Target Database (TTD) and GeneCards database. A protein-protein interaction (PPI) network was constructed using the STRING database. Furthermore, GO function and KEGG enrichment analysis of the targets were analyzed using DAVID tools. Subsequently, the AutoDock software for molecular docking was adopted to verify the above network pharmacology analysis results between the active components and key targets. Finally, an SCI rat model animal validation experiment was assessed to verify the reliability of the network pharmacology results. RESULTS: There were 7 active ingredients identified in CR and 246 SCI-related targets were collected. Then, 4 core nodes (ALB, AKT1, MAPK1, and EGFR) were discerned via construction of a PPI network of 111 common targets. The KEGG enrichment analysis results indicated that the Ras signaling pathway, estrogen signaling pathway, and vascular endothelial growth factor (VEGF) signaling pathway were enriched in the development of SCI. The results of molecular docking demonstrated that the effects of CR have a strong affinity with the 4 pivotal targets. Experimental validation in a rat model showed that CR could effectively improve the recovery of motor function and mechanical pain threshold after SCI. CONCLUSIONS: In summary, it revealed the mechanism of CR treatment for SCI involve active ingredients, targets and signaling pathways, providing a scientific basis for future investigations into the mechanism underlying CR treating for SCI.

Simultaneous determination of 35 constituents and elucidation of effective constituents in a multi-herb Chinese medicine formula Xiaoer-Feire-Kechuan / 药物分析学报

Xiaoer-Feire-Kechuan (XFK) is an 11-herb Chinese medicine formula to treat cough and pulmonary inflammation.The complicated composition rendered its chemical analysis and effective-component elucidation.In this study,we combined quantitative analysis and bioactivity test to reveal the anti-inflammatory constituents of XFK.First,UPLC-DAD and UHPLC/Q-Orbitrap-MS methods were estab-lished and validated to quantify 35 analytes (covering 9 out of 11 herbs) in different XFK formulations.Parallel reaction monitoring mode built in Q-Orbitrap-MS was used to improve the sensitivity and selectivity.Then,anti-inflammatory activities of the 35 analytes were analyzed using in vitro COX-2 inhibition assay.Finally,major analytes forsythosides H,I,A (8-10),and baicalin (15) (total contents varied from 21.79 to 91.20 mg/dose in different formulations) with significant activities (inhibitory rate ≥ 80％) were proposed as the anti-inflammatory constituents of XFK.The present study provided an effective strategy to discover effective constituents of multi-herb formulas.

Downregulation of OsAGO17 by artificial microRNA causes pollen abortion resulting in the reduction of grain yield in rice

Background: Pollen development is an important reproductive process that directly affects pollen fertility and grain yield in rice. Argonaute (AGO) proteins, the core effectors of RNA-mediated silencing, play important roles in regulating plant growth and development. However, few AGO proteins in rice were reported to be involved in pollen development. In this study, artificial microRNA technology was used to assess the function of OsAGO17 in pollen development. Results: In this study, OsAGO17, a rice-specific gene, was specifically expressed in rice pollen grains, with the highest expression in uninucleate microspores. Downregulation of OsAGO17 by artificial microRNA technology based on the endogenous osa-miRNA319a precursor was successfully achieved. It is found that downregulation of OsAGO17 could significantly affect pollen fertility and cause pollen abortion, thus suggesting that OsAGO17 functions in rice pollen development. In addition, the downregulation of OsAGO17 mainly caused a low seed-setting rate, thereby resulting in the reduction of grain yield, whereas the downregulation of OsAGO17 did not significantly affect rice vegetative growth and other agricultural traits including number of florets per panicle, number of primary branch per panicle, and 100-grain weight. Furthermore, the result of subcellular localization analysis indicated that the OsAGO17 protein was localized to both the nucleus and the cytoplasm. Conclusion: These results represent the first report of the biological function for OsAGO17 in rice and indicate that OsAGO17 may possibly play crucial regulatory roles in rice pollen development. It helps us to better understand the mechanism of pollen development in rice.

Cellular Phenotypic Analysis of Macrophage Activation Unveils Kinetic Responses of Agents Targeting Phosphorylation.

Macrophages are highly plastic cells, which serve as sentinels of the host immune system due to their ability to recognize and respond to microbial products rapidly and dynamically. Appropriate regulation of macrophage activation is essential for pathogen clearance or preventing autoimmune diseases. However, regularly used endpoint assays for analyzing macrophage functions have the limitations of being static and non-high throughput. In this study, we introduced a real-time and convenient method based on changes in cellular impedance that are detected by microelectronic biosensors. This new method can record the time/dose-dependent cell response profiles (TCRPs) of macrophages in real time and generates physiologically relevant data. The TCRPs generated from classically interferon-γ/lipopolysaccharide-activated macrophages showed considerable consistency with the data generated from standard endpoint assays. We further explored this approach by using it for global screening of a library of protein tyrosine kinase/phosphatase (PTK/PTP) inhibitors to investigate their impact on macrophage activation. Collectively, our findings suggest that the cellular impedance-based assay provides a promising approach for dynamically monitoring macrophage functions in a convenient and high-throughput manner.

Effects of quercetin on hedgehog signaling in chronic myeloid leukemia KBM7 cells.

OBJECTIVE: To investigate the effects of quercetin on Hedgehog (Hh) signaling in chronic myeloid leukemia KBM7 cells. METHODS: The KBM7 cells were treated with 50, 100 and 200 µmol/L quercetin for 48 h respectively. And then the trypan blue assay was used to examine the proliferative inhibition of quercetin. Apoptotic cells and cell cycle were measured by flow cytometry. The mRNA and protein expression were detected by quantitative real-time polymerase chain reaction (PCR) and Western blot, respectively. RESULTS: Quercetin significantly inhibited KBM7 cell proliferation, induced cell apoptosis, and blocked cell cycle at G1 phase, which were in dose-dependent manners. The mRNA and protein expression of Smoothened and Glioma1 (Gli1), the members of Hh pathway decreased after treatment with quercetin. The Bcl-2 and Cyclin D1, targets of Hh signaling, also decreased after treatment with quercetin, respectively. Quercetin also could increase p53 and Caspase-3 expression. Bcr-abl mRNA copies decreased, but no changes of phosphorylated Bcr-abl and Bcr-abl proteins were observed, after treatment with quercetin. CONCLUSION: Quercetin could inhibit Hh signaling and its downstream targets in the KBM7 cells. And it might be one of mechanisms of inducing apoptosis and inhibiting cell cycle by quercetin.

Glyphosate effects on the gene expression of the apical bud in soybean (Glycine max).

Glyphosate is a broad spectrum, non-selective herbicide which has been widely used for weed control. Much work has focused on elucidating the high accumulation of glyphosate in shoot apical bud (shoot apex). However, to date little is known about the molecular mechanisms of the sensitivity of shoot apical bud to glyphosate. Global gene expression profiling of the soybean apical bud response to glyphosate treatment was performed in this study. The results revealed that the glyphosate inhibited tryptophan biosynthesis of the shikimic acid pathway in the soybean apical bud, which was the target site of glyphosate. Glyphosate inhibited the expression of most of the target herbicide site genes. The promoter sequence analysis of key target genes revealed that light responsive elements were important regulators in glyphosate induction. These results will facilitate further studies of cloning genes and molecular mechanisms of glyphosate on soybean shoot apical bud.

Identification of novel acyl-ACP thioesterase gene ClFATB1 from Cinnamomum longepaniculatum.

A putative fatty acyl-acyl carrier protein (acyl-ACP) thioesterase (thioesterase) full-length cDNA sequence named as ClFATB1 was obtained from the seed cDNA library of Cinnamomum longepaniculatum by the SMART-RACE method. The novel gene encodes a protein of 382 amino acid residues with close homology to fatty acid thioesterase type B (FATB) enzymes of other plants, with two essential residues (His285 and Cys320) for thioesterase catalytic activity. The gene was transcribed in all tissues of C. longepaniculatum, the highest being in seeds. Recombinant ClFATB1 in Escherichia coli had higher specific activities against saturated 16:0- and 18:0-ACPs than on unsaturated 18:1-ACP. Overexpression of ClFATB1 in transgenic tobaccos upregulated thioesterase activities of crude proteins against 16:0-ACP and 18:0-ACP by 20.3 and 5.7%, respectively, and resulted in an increase in the contents of palmitic and stearic acids by 15.4 and 10.5%, respectively. However, ectopic expression of this gene decreased the substrate specificities of crude proteins to unsaturated 18:1-ACP by 12.7% in transgenic tobacco and lowered the contents of oleic, linoleic, and linolenic acids in transgenic leaves. So ClFATB1 would potentially upregulate the synthesis of saturated fatty acids and downregulate unsaturated ones in the fatty acid synthesis pathway of plants.

MicroRNA profiles and their control of male gametophyte development in rice.

Plant microRNAs (miRNAs) act as negative regulators of gene expression by slicing target transcripts or inhibiting translation. A number of miRNAs play important roles in development. In order to investigate the potential function of miRNAs during male gametogenesis in rice, we obtained both gene and small RNA expression profiles by combining microarray and high-throughput sequencing technologies. From the microarray datasets, 2,925 male gametophyte-specific genes were identified, including 107 transcription factors and three significant Argonaute genes (AGO12, AGO13, and AGO17). From the sRNA-Seq datasets, 104 unique miRNAs (miRus) were identified, including 47 known miRus and 57 novel miRus; interestingly, most of the new miRus are pollen-specific and not conserved among species. Furthermore, an interactive network of miRNA-target was constructed based on the two datasets. By employing enrichment analysis, the miRNA-regulated targets were found to be involved in both the up and down pathways, but predominantly in the down pathways, including 37 GO biological processes and 32 KEGG pathways. These findings indicate that miRNAs play a broad regulatory role during male gametophyte development in rice.

[Study on needling Ying method for treatment of sore throat].

To explore the mechanism of needling Ying method for treatment of sore throat. By the analysis of pathogenesis of sore throat, the authors think the key of its pathogenesis is stagnation of pathogenic factors such as hotness and phlegm accumulating, and meridian-vessel obstruction in the throat is its meridian foundation. There are several meridians passing through the throat, so the throat is closely related to viscera and meridians, and stagnation of pathogenic factors such as hotness and phlegm accumulating in the throat lead to sore throat when exogenous pathogenic factors invading or dysfunction of viscera and meridians. The treatment of needling Ying at local throat or combined with corresponding meridian point selection can dredge collaterals, dispel pathogenic factors, remove pathogenic factors to dispel swelling, resolve phlegm and dissipate stagnation and harmonize yin and yang, so as to relieve sore throat. In conclusion, needling Ying method is an important method in the treatment of sore throat.

[Leaf nitrogen and phosphorus stoichiometry across 654 terrestrial plant species in NSTEC].

Stoichiometry of leaf N and P is a characteristic of plant to adapt to environment, and can provide data for process-based modeling at large scale. Leaf nitrogen and phosphorus stoichiometry of NSTEC terrestrial plants were studied based on a NSTEC data set including leaf nitrogen and phosphorus observations for 654 plant species at 168 sites. The results showed that leaf nitrogen and phosphorus stoichiometry in NSTEC exhibited large variations, primarily ranging 2.17-52.61 mg x g(-1) for N, 0.10-10.27 mg x g(-1) for P, 1.7-74.6 for N/P ratio. Geometric means for all plant species were 17.55 mg x g(-1), 1.28 mg x g(-1) and 13.5, respectively. Leaf P of NSTEC (ever across China) was lower than global level, therefore plant growth was more limited by P in China region comparing to global environment. For all functional groups, the difference of leaf N was largest, but that of N/P ratio was smallest, since leaf N and P closely related in most of functioning groups; for leaf N and P of the different phylogenic functional groups, the difference was largest for farthest relative (seed vs fern), least for closest relative (monocotyledon vs. dicotyledon) . There were obvious correlation between leaf N (or P) and latitude (or mean annual temperature, MAT). Leaf N and P significantly increased with latitude increasing (or MAT decreasing), but the relationship between N/P ratio and latitude (or MAT) wasn't significant (p = 0.386 and p = 0.342), and the reason maybe include, leaf N and P had the same tendencies and large variations, and region in this research was smaller than global research.

[Clinical observation on acupuncture kinetotherapy for acute simple laryngitis of wind-heat type].

OBJECTIVE: To observe clinical therapeutic effect of acupuncture kinetotherapy for acute simple laryngitis of wind-heat type and to probe the mechanism. METHODS: Eighty cases were randomly divided into a test group and a control group, 40 cases in each group. The test group were treated with acupuncture kinetotherapy (Acupuncture was given at Kaiyin point No. 1 combined with deep respiratory movement of the glottis), once daily. The control group were treated with ultrasonic aerosol inhalation of 0.9% sodium chloride injection, gentamicin sulfate injection 80 000 U and Dexamethasone injection 5mg, twice a day. The treatment was given for 5 days for the 2 groups. RESULTS: The cured and markedly effective rate was 90.0% in the test group and 82.5% in the control group with no significant difference between the two groups (P>0.05); in the test group there was a very significant difference before and after treatment in the cumulative score of symptoms and the acoustic parameters (P<0.01). CONCLUSION: Acupuncture kinetotherapy has obvious therapeutic effect on acute simple laryngitis of wind heat type, with no toxic and adverse effects.

[Acupuncture treatment of vocal nodule and evaluation of the therapeutic effect by computerized acoustics assay].

OBJECTIVE: To evaluate the therapeutic effect of acupuncture at point Kaiyin No. 1 on vocal nodule. METHODS: Eighty cases of vocal nodule were randomly divided into an acupuncture group of 40 cases, a Chinese herb group of 20 cases and a western medicine group of 20 cases. The acupuncture group were treated with acupuncture at main point Kaiyin No. 1, the Chinese herb group with oral administration of Jinsang Sanjie Pill and the Western medicine group with spray inhalation of antibiotics and hormones. Their therapeutic effects and parameters of acoustics before and after treatment were observed. RESULTS: The total effective rate was 92.5% in the acupuncture group, significantly better than 85.0% in the Chinese herb group and 85.0% in the Western medicine group (P < 0. 01); the acupuncture group in improvement of parameters of acoustics was significantly superior to other two groups (both P < 0.01). CONCLUSION: Acupuncture at point Kaiyin No. 1 has significant therapeutic effect on vocal nodule with no adverse effect.

[The identification and cloning of human M961 full-length cDNA and its splicing isoform].

OBJECTIVE: To identify and clone the gene encoding human M96 gene and study its expression spectrum in several blood cell lines. METHODS: According to the sequence of human EST which was highly homologous to the mouse M96 gene, primers used for library screening were synthesized, then the human adult testis and fetal brain cDNA library were screened. The gene was analyzed by making use of BLAST and CLUSTAL W, and its expression spectrum was studied by multiple-cell lines Northern blot analysis. The expression change of M961 in cell differentiation was observed by use of K562 cell line induced by hemin. RESULTS: Two cDNA clones encoding human M96 gene were isolated, identified and named as M961, and M962. They were found to be isoforms of each other. Northern, blot showed that M961 gene was expressed highly in CEM, Hel, Dami and K562 cell lines. However, during K562 cell line differentiation, process the expression of M961 elevated only slightly. CONCLUSIONS: M961 gene was expressed highly in pluripotent cell lines with erythrocytic and megakaryocytic potentials.