[Sequence analysis and identification of a novel HLA-DPB1*02:01:69 allele by third-generation sequencing].

OBJETIVE: To analyze the sequence of a novel HLA-DPB1 allele in an individual. METHODS: A individual identified from the database of blood donors for matched platelet transfusion at the Blood Center of Zhejiang Province in May 2022 was selected as the study subject. HLA genotype of the individual was determined by next-generation sequencing (NGS) on an Ion Torrent S5 platform. The sequence of the HLA-DPB1 locus was also determined by NGS on an Illumina Miseq platform and third-generation sequencing using Oxford Nanopore MinION. This study was approved by the Blood Center of Zhejiang Province (Ethics No. 2021-001). RESULTS: A novel HLA-DPB1*02 allele was identified in the specimen, for which the closest genotype was HLA-DPB1*02:new,17:01:01G, with the variant located in exon 3. Meanwhile, the NGS also revealed a novel HLA-DPB1*17 allele, with the closest genotype being HLA-DPB1*02:01,17:new. Both the HLA-DPB1*17:01:01:01 and HLA-DPB1*02 alleles were identified by third-generation sequencing. Compared with the HLA-DPB1*02:01:02:01 allele, the novel allele had a G>A variation at position 369 in the exon 3, which however did not result in amino acid change. CONCLUSION: A novel HLA-DPB1 allele has been identified and validated by both NGS and TGS, which has been named as HLA-DPB1*02:01:69 by the World Health Organization Committee on Nomenclature of Factors of the HLA System.

Cerebellar Fastigial Nucleus Stimulation in a Chronic Unpredictable Mild Stress Rat Model Reduces Post-Stroke Depression by Suppressing Brain Inflammation via the microRNA-29c/TNFRSF1A Signaling Pathway.

BACKGROUND We previously reported that cerebellar fastigial nucleus stimulation reduced post-stroke depression in a rat model by reducing inflammation. This study aimed to investigate the molecular inflammatory signaling pathways associated with cerebellar fastigial nucleus stimulation in an established rat model of post-stroke depression. MATERIAL AND METHODS Twenty-four Sprague-Dawley rats included a sham group (N=6), an untreated stroke group (N=6), an untreated post-stroke depression model group (PSD) (N=6), and the model group treated with cerebellar fastigial nucleus stimulation (FNS) (N=6). The rat stroke model involved occlusion of the middle cerebral artery occlusion (MCAO). Post-stroke depression model was established using chronic unpredictable mild stress treatment and was verified using an open field test. Real-time polymerase chain reaction (PCR) and Western blot compared expression levels of microRNA-29c (miR-29c), miR-676, TNFRSF1A, tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6 and IL-1ß in cerebellar tissue. U251 human glioblastoma cells and SH-SY5Y human neuroblastoma cells were studied in vitro. RESULTS Cerebellar fastigial nucleus stimulation reduced behaviors associated with depression in the rat model, upregulated the expression of miR-29c, and reduced the expression of TNFRSF1A and inflammatory cytokines, and mildly reduced neuronal apoptosis. Bioinformatics data analysis identified a regulatory relationship between miR-29c and TNFRSF1A. SH-SY5Y cells treated with a miR-29c mimic, or TNFRSF1A short interfering RNA (siRNA), identified a negative regulatory relationship between TNFRSF1A and miR-29c. CONCLUSIONS In a rat model, cerebellar fastigial nucleus stimulation reduced the expression of TNFRSF1A by upregulating miR-29c expression, which suppressed the expression of inflammatory cytokines, resulting in reduced severity of post-stroke depression.

Fungal community structure shifts in litter degradation along forest succession induced by pine wilt disease.

Fungi play a crucial role in decomposing litter and facilitating the energy flow between aboveground plants and underground soil in forest ecosystems. However, our understanding how the fungal community involved in litter decomposition responds during forest succession, particularly in disease-driven succession, is still limited. This study investigated the activity of degrading enzyme, fungal community, and predicted function in litter after one year of decomposition in different types of forests during a forest succession gradient from coniferous to deciduous forest, induced by pine wilt disease. The results showed that the weight loss of needles/leaves and twigs did not change along the succession process, but twigs degraded faster than needles/leaves in both pure pine forest and mixed forest. In pure pine forest, peak activities of enzymes involved in carbon degradation (ß-cellobiosidase, ß-glucosidase, ß-D-glucuronidase, ß-xylosidase), nitrogen degradation (N-acetyl-glucosamidase), and organic phosphorus degradation (phosphatase) were observed in needles, which subsequently declined. The fungal diversity and evenness (Shannon's diversity and Shannon's evenness) dropped in twig from coniferous forest to mixed forest during the succession. The dominant phyla in needle/leaf and twig litters were Ascomycota (46.9%) and Basidiomycota (38.9%), with Lambertella pruni and Chalara hughesii identified as the most abundant indicator species. Gymnopus and Desmazierella showed positively correlations with most measured enzyme activities. Functionally, saprotrophs constituted the main trophic mode (47.65%), followed by Pathotroph-Saprotroph-Symbiotroph (30.95%) and Saprotroph-Symbiotroph (10.57%). The fungal community and predicted functional structures in both litter types shifted among different forest types along the succession. These findings indicate that the fungal community in litter decomposition responds differently to disease-induced succession, leading to significant shifts in both the fungal community structure and function.

[Effect of substrate temperature on structure and photoluminescence of ZnMgO films].

ZnMgO films were deposited on quartz glass substrates by the ultrasonic spray pyrolysis at different substrate temperatures (450-550 degrees C). The structural, surface morphological and optical properties of the samples were investigated by X-ray diffraction (XRD), scanning electron microscope (SEM) and photoluminescence (PL) spectroscopy. The results demonstrate that the substrate temperature has important effect on structural and optical characteristics. All the films have hexagonal wurtzite polycrystalline structures and the c-axis preferential orientation has an optimum temperature of 530 degrees C. The sample prepared at this temperature owns uniform grain size, smooth surface morphology and better crystalline quality. The width of deep-level emission decreases and the near band edge (NBE) ultraviolet emission peak appears with the increase in temperature by the PL spectrum. When the temperature arrives to 530 degrees C, a distinct NBE emission peak can be observed at 374. 5 nm, while the deep level emission is almost undetectable.

[Analysis of Gene Recombination between HLA-B and -DRB1, HLA-DQB1 and -DPB1 Loci].

OBJECTIVE: To investigate the recombinations within the human leukocyte antigen (HLA) region in two families. METHODS: Genomic DNA was extracted from the peripheral blood specimens of the different family members. HLA-A, -B, -C, -DRB1, -DQB1 and -DPB1 loci were genotyped using polymerase chain reaction-sequence specific oligonucleotide probing technique (PCR-SSO) and next-generation sequencing technique. HLA haplotype was determined by genetic analysis of the pedigree. RESULTS: The haplotypes of HLA-A*11:01~C*03:04~B*13:01~DRB1*12:02~DQB1*03:01~DPB1*05:01:01G and HLA-A*03:01~C*04:01~B*35:03~DRB1*12:01~DQB1*03:01~DPB1*04:01:01G in the family 1 were recombined between HLA-B and HLA-DRB1 loci, which formed the haplotype of HLA-A*11:01~C*03:04~B*13:01~DRB1* 12:01~DQB1*03:01~DPB1*04:01:01G. The haplotypes of HLA-A *02:06~C*03:03~B*35:01~DRB1*08:02~DQB1*04:02~ DPB1*13:01:01G and HLA-A *11:01~C*07:02~B*38:02~DRB1*15:02~DQB1*05:01~DPB1*05:01:01G in the family 2 were recombined between HLA-DQB1 and HLA-DPB1 loci, which formed the haplotype of HLA-A*02:06~C*03:03~B*35:01~ DRB1*08:02~DQB1*04:02~DPB1*05:01:01G. CONCLUSION: The gene recombination events between HLA-B and -DRB1, HLA-DQB1 and -DPB1 loci were found respectively in two Chinese Han families.

A multitask classification framework based on vision transformer for predicting molecular expressions of glioma.

PURPOSE: The purpose of this study is to develop a Vision Transformer model with multitask classification framework that is appropriate for predicting four molecular expressions of glioma simultaneously based on MR imaging. MATERIALS AND METHODS: A total of 188 glioma (grades II-IV) patients with an immunohistochemical diagnosis of IDH, MGMT, Ki67 and P53 expression were enrolled in our study. A Vision Transformer (ViT) model, including three independent networks based on T2WI, T1CWI and T2 + T1CWI (T2-net, T1C-net and TU-net), was developed for the prediction of four glioma molecular expressions simultaneously. To evaluate the model performance, the accuracy rate, recall, precision, F1-score, and area under the receiver operating characteristic curve (AUC) were calculated. RESULTS: The proposed ViT model achieved high accuracy in predicting IDH, MGMT, Ki67 and P53 expression in gliomas. Among the three networks using the ViT model, TU-net achieved the best results with the highest values of accuracy (range, 0.937-0.969), precision (range, 0.949-0.972), recall (range, 0.873-0.991), F1-score (range, 0.910-0.981) and AUC (range, 0.976-0.984). Comparisons were also made between our ViT model and convolutional neural network (CNN)-based models, and the proposed ViT model outperformed the existing CNN-based models. CONCLUSION: Vision Transformer is a reliable approach for the prediction of glioma molecular biomarkers and can be a viable alternative to CNNs.

A new species (Begoniagiganticaulis) of Begoniaceae from southern Xizang (Tibet) of China.

Begoniagiganticaulis, a huge new species in Begoniasect.Platycentrum of Begoniaceae from southern Xizang (Tibet) of China, is described. Morphologically, it is mostly similar to B.longifolia and B.acetosella, but clearly differs from the former mainly by its dioecious and taller plants, sparse hairs on abaxial veins, longer inflorescence, unique shape of fruits, and differs from the latter mainly by its late and longer flowering time, 6-tepals of female flower and 3-loculed ovary. The phylogenetic analyses also support the separation of the new species from other taxa. Based on the current data, its conservation status is assigned to Endangered (B2a) according to the IUCN Red List Categories and Criteria.

Begonia catbensis (sect. Coelocentrum, Begoniaceae), a new species from northern Vietnam.

Begonia catbensis, a new species in Begonia sect. Coelocentrum is described and illustrated. The new species was discovered in lowland limestone hills at Cat Ba National Park and can be easily distinguished from all its congeners by having dendritic hairs on the petiole, adaxial veins and stipules, fimbriate bracts and bracteoles, dense conical bullae on the upper surface of the leaf blade, two tepals in the pistillate flowers and a glabrescent ovary with verrucose wings. Based on IUCN Criteria, the species is currently assessed as "Endangered" (D).

Extended expression of B-class MADS-box genes in the paleoherb Asarum caudigerum.

Asarum caudigerum (Aristolochiaceae) is a paleoherb species that is important for research in origin and evolution of angiosperm flowers due to its basal position in the angiosperm phylogeny. In this study, a subtracted floral cDNA library from floral buds of A. caudigerum was constructed and cDNA arrays by suppression subtractive hybridization were generated. cDNAs of floral buds at different stages before flower opening and of leaves at the seedling stage were used. The macroarray analyses of expression profiles of isolated floral genes showed that 157 genes out of the 612 unique ESTs tested revealed higher transcript abundance in the floral buds and uppermost leaves. Among them, 78 genes were determined to be differentially expressed in the perianth, 62 in the stamens, and 100 genes in the carpels. Quantitative real-time PCR of selected genes validated the macroarray results. Remarkably, APETALA3 (AP3) B-class genes isolated from A. caudigerum were upregulated in the perianth, stamens and carpels, implying that the expression domain of B-class genes in this basal angiosperm was broader than those in their eudicot counterparts.

[Effects of shading on photosynthetic characteristics and chlorophyll fluorescence parameters of four Corydalis species]. / 遮阴对4ç§ç´«å ‡å±žæ¤ç‰©å ‰åˆç‰¹æ€§å’Œå¶ç»¿ç´ è§å ‰å‚æ•°çš„å½±å“.

We examined the effects of five shading treatments (0, 20%, 40%, 60% and 80% shading) on chlorophyll content, photosynthetic characteristics, and chlorophyll fluorescence chara-cteristics of four Corydalis species (C. incisa, C. decumbens, C. edulis and C. pallida) in a pot experiment. The results showed that the contents of chlorophyll a, chlorophyll b and chlorophyll (a+b) increased with the increment of shading, with that of C. incisa reaching the maximum under 80% shading treatment and that of C. decumbens, C. edulis and C. pallida reaching a maximum under 60% shading treatment. In contrast, chlorophyll a/b, light saturation point, light compensation point and dark respiration rate decreased with increasing shading. Among the four Corydalis species, C. incisa reached up to the maximum chlorophyll fluorescence parameters under 80% shading treatment, and C. decumbens, C. edulis and C. pallida reached the maximum at 60% shading treatment. The shade tolerance of four species was as follows: C. incisa ＞ C. decumbens ＞ C. edulis ＞ C. pallida. C. incisa under 80% shading treatment and C. decumbens, C. edulis, C. pallida at 60% shading treatment had the highest light energy utilization and photosynthetic capacity, which would facilitate their growth.

[A clinical study of chromosome translocations in extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue in Chinese patients].

OBJECTIVE: To investigate the genetic aberrations in extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT) lymphomas from different sites of the body in Chinese patients. METHODS: Two hundred and seventeen paraffin-embedded MALT lymphoma specimens from 11 major sites were studied with interphase fluorescence in situ hybridization (FISH) to detect t (11; 18) (q21; q21)/API2-MALT1, t (1; 14) (p22; q32)/IGH-BCL10, (14; 18) (q32; q21)/IGH-MALT1 and BCL6 gene involved chromosome translocations. RESULTS: These translocations were mutually exclusive and detected in 21% (46/217) of the cases, including t (11; 18) (q21; q21) API2-MALT1 13% (29/217), t (1; 14) (p22; q32) IGH-BCL10 in 1% (3/217), t (14; 18) (q32; q21) IGH-MALT1 1% (2/217), BCL6 involved translocation in 2% (4/217) and IGH-unknown translocation partner in 4% (8/217). t (11; 18) (q21; q21) API2-MALT1 was found with the highest frequency in MALT lymphoma from lungs (47%, 8/17) and small intestine (29%, 4/14), followed by salivary gland (17%, 1/6), stomach (14%, 12/84) and ocular adnexae (6%, 4/68). t (1; 14) (p22; q32) was only detected in lungs (12%, 2/17) and stomach (1%, 1/84). t (14; 18) (q32; q21) was mainly detected in lungs (6%, 1/17) and ocular adnexae (2%, 1/68). BCL6 gene involved translocation was detected in salivary gland (17%, 1/6) and stomach (4%, 3/84). CONCLUSIONS: It is demonstrated that the four translocations occur with markedly variable frequencies in MALT lymphoma of different sites in Chinese patients. The distributions of these chromosome translocations in Chinese patients are slightly different from those reported in western patients.

[Relationship between primary ocular adnexal mucosa-associated lymphoid tissue lymphoma and eye infection].

OBJECTIVE: To study the role of pathogenic microorganisms commonly associated with chronic eye disease, including Chlamydia psittaci, Chlamydia trachomatis, Chlamydia pneumoniae, herpes simplex virus (HSV) type 1 and type 2, and adenovirus type 8 and type 19, in the development of primary ocular adnexal mucosa-associated lymphoid tissue (MALT) lymphoma in Chinese patients. METHODS: Sixty-eight archival cases of primary ocular adnexal lymphoproliferative lesions, including 38 cases of MALT lymphoma, 3 cases of non-MALT lymphoma and 27 cases of chronic inflammation, were enrolled into the study. DNA was extracted from the paraffin-embedded tissue samples. The presence of DNA of C. psittaci, C. trachomatis, C. pneumoniae, HSV type 1, HSV type 2, adenovirus type 8 and adenovirus type 19 were analyzed by multiplex touchdown enzyme time-release polymerase chain reaction (TETR-PCR). RESULTS: All of the specimens yielded PCR products of over 100 base pairs and were thus suitable for TETR-PCR screening of infectious agents. The prevalence of DNA of C. psittaci, C. trachomatis and adenovirus type 19 were 0 in MALT lymphoma, non-MALT lymphoma and chronic inflammation. There were 2 cases positive for C. pneumoniae DNA, amongst the 38 cases of MALT lymphoma studied (5.3%, 2/38). HSV type 1, HSV type 2 and adenovirus type 8 DNA was found in each of the 3 patients with chronic inflammation. CONCLUSION: The study indicates that C. psittaci, C. trachomatis, C. pneumoniae, HSV type 1, HSV type 2, adenovirus type 8 and adenovirus type 19 probably play little role in the pathogenesis of ocular adnexal MALT lymphoma in Chinese patients.

Role of intestinal microbiota and metabolites in inflammatory bowel disease.

OBJECTIVE: The metabolites produced by the gut microbiota are of interest to scientists. The objective of this review was to provide an updated summary of progress regarding the microbiota and their metabolites and influences on the pathogenesis of inflammatory bowel disease (IBD). DATA SOURCES: The author retrieved information from the PubMed database up to January 2018, using various combinations of search terms, including IBD, microbiota, and metabolite. STUDY SELECTION: Both clinical studies and animal studies of intestinal microbiota and metabolites in IBD were selected. The information explaining the possible pathogenesis of microbiota in IBD was organized. RESULTS: In IBD patients, the biodiversity of feces/mucosa-associated microbiota is decreased, and the probiotic microbiota is also decreased, whereas the pathogenic microbiota are increased. The gut microbiota may be a target for diagnosis and treatment of IBD. Substantial amounts of data support the view that the microbiota and their metabolites play pivotal roles in IBD by affecting intestinal permeability and the immune response. CONCLUSIONS: This review highlights the advances in recent gut microbiota research and clarifies the importance of the gut microbiota in IBD pathogenesis. Future research is needed to study the function of altered bacterial community compositions and the roles of metabolites.

Lysimachia fanii, a new species of Primulaceae from limestone area of Guangxi, China.

Lysimachia fanii, a new species of Lysimachia (Subgen. Idiophyton, Primulaceae), is described and illustrated from Guangxi, China based on morphological and molecular data. Lysimachia fanii differs from L. verbascifolia, L. rupestris and L. alpestris mainly by the habit being nearly rosulate, leaves congested at the apex of the rhizome, leaf blades spatulate to narrowly oblanceolate and flowers solitary. Phylogenetic analyses supported L. verbascifolia as sister to L. fanii. This new species is endemic to limestone areas in Liucheng county of Guangxi, China.

The complete plastid genome sequence of Begonia guangxiensis.

Begonia guangxiensis was assessed as endangered according to Red List of Chinese Plants. In this study, we described the complete plastid genome of B. guangxiensis. The plastid genome sequence of B. guangxiensis is 157,648 bp in size, having a large single-copy region (LSC) of 86,514 bp, a small single-copy region (SSC) of 18,076 bp, and two inverted repeat (IR) regions of 26,529 bp. The complete plastid genome of B. guangxiensis encoding 112 unique genes including 79 protein-coding genes, 29 tRNA genes, and four rRNA genes. The overall GC content of the plastid genome is 35.9%. Phylogenetic analysis confirmed that B. guangxiensis is closely related to B. varipeltata.

[Anaplastic lymphoma kinase gene abnormality and the expression of its fusion protein in primary systemic anaplastic large cell lymphoma].

OBJECTIVE: To study the expression of anaplastic lymphoma kinase (ALK) and chromosome breakage of the anaplastic lymphoma kinase (ALK) gene retrospectively and to investigate their possible value as indicators of prognosis in primary systemic anaplastic large cell lymphomas (S-ALCL). METHODS: Twenty-eight cases of S-ALCL were collected from the Lymphoma Lab, the Department of Pathology, Peking University Health Science Center and Beijing Children's Hospital. The morphologic characteristics were studied under light microscope, and essential immunohistochemical staininings (IHC) were performed and reviewed to confirm the diagnosis of S-ALCL. ALK-1 monoclonal antibody was used to assess ALK fusion protein expression, and EnVision method was used in IHC. Locus specific interphase fluorescence in situ hybridization (LSI-FISH) was also performed on the neoplastic cells using paraffin-embedded tissues to detect ALK gene abnormality. RESULTS: ALK-1 protein was expressed in 19 of the 28 cases. In 14 ALCL cases, ALK gene breakage was detected by LSI-FISH, using a dual-color break-apart ALK gene DNA (LSI-ALK) probe. Of the other 14 cases which did not show ALK gene breakage, 5 showed 2 copies of ALK gene as normal, and 9 showed multi-copies of ALK gene. Of all the 28 cases, 22 had complete follow-up materials. Sixteen survived and 6 died, their survival time ranged from 0.5 to 36.0 months, and the survival time on average was 12.8 months, cumulative proportion survival rate was 73.9% in the 1st year. Those cases showing multi-copies of ALK gene might have the worst outcome, with only 47.6% of cumulative proportion survival rate in the 1st year. CONCLUSION: IHC detection for ALK fusion protein is important to the diagnosis of S-ALCL. ALK gene breakage detected by interphase LSI-FISH might not be always consistent with abnormal expression of ALK fusion protein. Complex abnormalities of ALK gene exist in S-ALCL cases, and different types of ALK gene might lead to different clinical outcome. Those cases with multi-copies of ALK gene probably have the poorest prognosis.

[Primary ocular adnexal lymphoproliferative lesions: clinicopathologic features and genetic alterations].

OBJECTIVE: To investigate clinicopathological and genetic characteristics of primary ocular adnexal lymphoproliferative lesions. METHODS: Clinical, morphological and immunohistochemical features of 37 archival cases of primary ocular adnexal lymphoproliferative lesions were studied including 5 cases of reactive lymphoid hyperplasia and 32 lymphomas retrospectively. Classification of the lymphomas were made according to the WHO classification of tumors of haematopoietic and lymphoid tissues. All cases were studied by interphase fluorescence in situ hybridization (FISH) using dual color break apart probes of IgH, MALT1, bcl-6, c-Myc, bcl-2, CCND1, bcl-10, and FOXP1 for detection of chromosomal aberrations involving IgH, MALT1, bcl-6, c-Myc, bcl-2, cyclinD1, bcl-10 and FOXP1 genes, respectively. FISH with IgH / bcl-2 dual color dual fusion probe was used for detection of t(14;18)(q32;q21)/IgH-bcl-2. CEP18 spectrum orange probe was used for detection of aneuploidy of the chromosome 18. RESULTS: Among 32 cases of lymphomas, 28 cases (87.5%) were extranodal marginal zone B-cell lymphomas of mucosa associated lymphoid tissue (MALT lymphoma), 2 cases were follicular lymphoma (FL) and 2 cases diffuse large B cell lymphoma (DLBCL). Among the 28 cases of MALT lymphoma, chromosomal aberrations were found in 60.7% (17/28) by interphase FISH analysis. One case showed positive IgH break-apart signal with unknown partner. 16 cases showed three copies of different genes, of which, three copies of MALT1, bcl-6, and c-Myc were identified in 7 cases (25%), 12 cases (43%), and 2 cases (8%) of MALT lymphomas, respectively. In addition, 5 cases showed two genes including three copies of bcl-6 and MALT1 in 4 cases, and three copies of bcl-6 together with c-Myc in one case. Furthermore, all cases with three copies of MALT1 had trisomy 18. t(14;18)(q32;q21) was detected in both follicular lymphomas. Of the 2 DLBCL cases, one showed three copies of bcl-6 together with trisomy 18 and the other one showed three copies of bcl-6 together with IgH and c-Myc rearrangements. Chromosomal aberration was not found in all 5 cases of reactive lymphoid hyperplasia. CONCLUSIONS: The most common entity of primary ocular adnexal lymphomas is MALT lymphoma and FISH is helpful for their differential diagnosis and classification. Trisomy 18 and three copies of bcl-6 are common chromosomal aberrations in primary ocular adnexal MALT lymphomas.

[Comparative study on contents of amino acid and major and trace elements in Cornus officinalis before and after being processed].

OBJECTIVE: To investigate amino acid and inorganic element in cornus officinalis and study the effects of processed on contents of them. METHODS: The amino acid analysis with precolume derivatization was implemented by HPLC and the contents of element in the prevention were determined by ICP. RESULTS: cornus officinalis contained eighteen inorganic element and abundant K, Ca, Mg. After processed, except for Cu, Ba, Ni, all were increased and the contents of La and Ce were added remarkably. Sixteen amino acid were determined in cornus officinalis and rich asp and glu were measurated. The total content of amino acid decreased, but contents of lys, phe and etc. increased. After processed, the contents of met, his were destroyed and pro reduced, however the total content of amino acid increased. CONCLUSION: cornus officinalis contains many types of amino acid and inorganic element. Processing lead to the dissolved of inorganic element increase and change of contents of amino acid. Plenty K, asp and the increase of the contents of lys, leu, val, and etc. being necessary for mamkind may be one of mechanism of enganced effect on protecting liver and kidney.

Faecal and mucosal microbiota in patients with functional gastrointestinal disorders: Correlation with toll-like receptor 2/toll-like receptor 4 expression.

AIM: To investigate the intestinal luminal microbiota (LM) and mucosa-associated microbiota (MAM) in Chinese patients with functional gastrointestinal disorders (FGIDs) and examine the association between these communities and the expression of toll-like receptor (TLR) 2 and TLR4. METHODS: Thirty-two Chinese subjects who suffered from symptoms of FGIDs, as confirmed by gastroenterologists, were enrolled in this study. Fresh faecal samples and descending colonic mucosal biopsies were collected from the subjects before (faecal) and during (mucosal) flexible colonoscopy. For analysis of the samples, we performed high-throughput sequencing of the V3-V4 region of the 16S rRNA gene and reverse transcription (RT)-PCR to detect the expression of colonic TLR2 and TLR4. Differences in the stool and mucosal microbiota were examined and a correlation network analysis was performed. RESULTS: The microbiota of faecal samples was significantly more diverse and richer than that of the mucosal samples, and the LM and MAM populations differed significantly. TLR2 expression showed a significant positive correlation with TLR4 expression. In the MAM samples, the genera Faecalibacterium and Ruminococcus, which belong to the family Ruminococcaceae, were inversely correlated with TLR4 expression (r = -0.45817, P = 0.0083 and r = -0.5306, P = 0.0018, respectively). Granulicatella, which belongs to Carnobacteriaceae, and Streptococcus, which belongs to Streptococcaceae, were inversely correlated with TLR2 expression (r = -0.5573, P = 0.0010 and r = -0.5435, P = 0.0013, respectively). In the LM samples, examination at phylum, class, or order level revealed no correlation with TLR4 expression. Faecalibacterium, which belongs to Ruminococcaceae, and Streptococcus, which belongs to Streptococcaceae, were inversely correlated with TLR2 expression (r = -0.5743, P = 0.0058 and r = -0.3905, P = 0.0271, respectively). CONCLUSION: Microbial compositions of LM and MAM in Chinese patients with FGIDs are different. Expression of TLRs may be affected by the type of bacteria that are present in the gut.

Endoplasmic reticulum stress regulates proliferation, migration and invasion of human ovarian cancer SKOV3 cells through PI3K/AKT/mTOR signaling pathway.

OBJECTIVE: The study is to explore the role of tunicamycin-induced endoplasmic reticulum stress (ERS) in human ovarian cancer (OC) SKOV3 cells proliferation, migration and invasion by modulating the activity of PI3K/AKT/mTOR pathway. METHODS: The collected human OC SKOV3 cells were randomly separated into three groups: The control group, the Tun group (treated with tunicamycin to induce ERS) and the CHOP-si group (transfected with CHOP-siRNA before tunicamycin treatment). CCK-8 method was applied for testing cell proliferation, while flow cytometry was conducted to detect cell apoptosis. Scratch test and Transwell test were used to determine the level of cell migration and invasion, respectively. Western blotting was performed to determine the related proteins expressions in ERS and PI3K/AKT/mTOR pathway. RESULTS: The cell survival rate in the Tun group was enhanced than that in the CHOP-si group, both of which were declined with the passage of time. The cell apoptosis rate in the Tun group was increased compared to the CHOP-si group, both of which were significantly elevated. The horizontal migration distance and the number of invasive cells in the Tun and CHOP-si groups were inhibited; however, the horizontal migration distance and the number of invasive cells in the CHOP-si group were enhanced than that in the Tun group. In comparison with the control group, the expressions of CHOP and TRB3 were increased in the Tun group but decreased in the CHOP-si group. The PI3K, p-AKT and p-mTOR expressions were remarkably declined in the Tun group than those in the control group (P< 0.05). CONCLUSION: The study provides strong evidence that tunicamycin-induced ERS induces the apoptosis of human OC SKOV3 cells through inhibiting PI3K/AKT/mTOR signaling pathway.