Specific expression profile of follicular fluid-derived exosomal microRNAs in patients with diminished ovarian reserve.

BACKGROUND: Diminished ovarian reserve (DOR) is defined as a reduction in ovarian reserve and oocyte quality. The pathophysiology of DOR has not been completely explained as of yet. Scholars have uncovered a large number of exosomes that have been detected in follicular fluid, and exosomal miRNAs have been proven to play a critical role in controlling ovarian disorders and follicle formation. We focused on the expression profile of follicular fluid-derived exosomal microRNAs (miRNAs) and attempted to understand if their role is connected to the pathomechanism of DOR. METHODS: The follicular fluid-derived differentially expressed exosomal miRNAs (DEmiRs) between patients with DOR and those with normal ovarian function were investigated using the next-generation sequencing (NGS) method. The main metabolic and signaling pathways of DEmiRs were identified using the KEGG pathway database, disease ontology (DO) analysis, and gene ontology (GO) analysis. In the end, a Protein-Protein Interaction (PPI) network was built to search for exosomal miRNAs and their target genes that were potentially strongly connected with DOR. RESULTS: In comparison to normal controls, 52 DEmiRs were discovered in follicular fluid-derived exosomes of DOR patients, of which 19 were up-regulated and 33 were down-regulated (|log2(fold change) |>2, P < 0.05). GO, DO analysis, and the KEGG pathway database revealed that many of these DEmiRs have broad biological roles that are connected to ovarian function and disorders. The top ten DEmiRs in terms of expression were then chosen for miRNA-mRNA interaction analysis. Totally, 8 experimentally supported miRNAs (hsa-miR-1246, hsa-miR-483-3p, hsa-miR-122-5p, hsa-miR-130b-3p, hsa-miR-342-3p, hsa-miR-625-3p, hsa-miR-675-3p, and hsa-miR-134-5p) and 126 target genes were filtrated by utilizing Cytoscape software. The module analysis findings of the PPI network showed that the main module cluster with a score > 6.0 (MCODE score = 15) had six hub genes, including IGFR, VEGFA, KRAS, ERBB2, RHOA, and PTEN (MCODE score = 11.472). CONCLUSION: Our data suggested a special expression profile of follicular fluid-derived exosomal miRNAs in patients with DOR, which was probably correlated to ovarian dysfunction and follicle formation. These results may give a unique insight into a better understanding of the molecular process in the pathogenesis of DOR or other ovarian diseases.

OsPHT1;3 Mediates Uptake, Translocation, and Remobilization of Phosphate under Extremely Low Phosphate Regimes.

Plant roots rely on inorganic orthophosphate (Pi) transporters to acquire soluble Pi from soil solutions that exists at micromolar levels in natural ecosystems. Here, we functionally characterized a rice (Oryza sativa) Pi transporter, Os Phosphate Transporter-1;3 (OsPHT1;3), that mediates Pi uptake, translocation, and remobilization. OsPHT1;3 was directly regulated by Os Phosphate Starvation Response-2 and, in response to Pi starvation, showed enhanced expression in young leaf blades and shoot basal regions and even more so in roots and old leaf blades. OsPHT1;3 was able to complement a yeast mutant strain defective in five Pi transporters and mediate Pi influx in Xenopus laevis oocytes. Overexpression of OsPHT1;3 led to increased Pi concentration both in roots and shoots. However, unlike that reported for other known OsPHT1 members that facilitate Pi uptake at relatively higher Pi levels, mutation of OsPHT1;3 impaired Pi uptake and root-to-shoot Pi translocation only when external Pi concentration was below 5 µm Moreover, in basal nodes, the expression of OsPHT1;3 was restricted to the phloem of regular vascular bundles and enlarged vascular bundles. An isotope labeling experiment with 32P showed that ospht1;3 mutant lines were impaired in remobilization of Pi from source to sink leaves. Furthermore, overexpression and mutation of OsPHT1;3 led to reciprocal alteration in the expression of OsPHT1;2 and several other OsPHT1 genes. Yeast-two-hybrid, bimolecular fluorescence complementation, and coimmunoprecipitation assays all demonstrated a physical interaction between OsPHT1;3 and OsPHT1;2. Taken together, our results indicate that OsPHT1;3 acts as a crucial factor for Pi acquisition, root-to-shoot Pi translocation, and redistribution of phosphorus in plants growing in environments with extremely low Pi levels.

Primary extra-nodal non-Hodgkin lymphoma in buttock soft tissue: A rare case report.

RATIONALE: Primary extra-nodal non-Hodgkin lymphoma (PE-NHL) arising in the region of the buttocks is rare. After reviewing the literature from the last 20 years, we found only 3 reported lymphomas originating from soft tissue of the buttocks. In our case, positron emission tomography/computed tomography (PET/CT) was performed for the first time, both before and after treatment, to determine the initial stage of PE-NHL and the curative effects of treatment. PATIENT CONCERNS: We report the case of a 71-year-old woman who was admitted to our hospital due to pain, skin redness, rising skin temperature, and swelling in the right hip. DIAGNOSES: After an initial misdiagnosis of local infection, a histological examination and PET/CT were performed which revealed evidence of non-Hodgkin marginal zone B cell lymphoma of Ann Arbor stage II. INTERVENTIONS: Following unsuccessful treatment with cephalosporin, the patient was successfully treated with rituximab combined with cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) chemotherapy. OUTCOMES: Comparison of the PET/CT scans taken before and after treatment showed that the lesion size had decreased, as had the fluorodeoxyglucose (FDG) uptake seen in the subcutaneous tissue of the right buttock with standardized uptake value max (SUVmax) 11.6 versus 2.5, respectively. Subsequently, no relapse or distant metastasis has been detected. LESSONS: Young doctors should suspect PE-NHL in similar cases. PET/CT is valuable in the diagnosis and treatment of PE-NHL, as well as for accurately determining PE-NHL stage and aggressiveness.

The prognostic value of regional lymph node metastases in patients of Guangdong Province, China with differentiated thyroid cancer: A multicenter retrospective clinical study.

Although the prognostic value of nodal metastases in differentiated thyroid cancer remains controversial, it is of interest to evaluate and understand the different characteristics of predictive outcomes.A multicenter retrospective study was conducted in 215 untreated patients with differentiated thyroid cancer from July 1997 to July 2015 in 4 medical centers of Guangdong Province. A total of 107 patients with nodal metastases (group A) were compared to 108 patients without metastases (group B). The 5-year disease-free survival (DFS), overall survival (OS), and postoperative complications in both groups were calculated. Variables predictive of DFS and OS were evaluated in group A.The group A had lower 5-year DFS (69.16%, 11 months) and shorter median time of recurrence than those in group B (87.96%, 8.5 months, respectively, P < 0.001). The incidence of temporary hypoparathyroidism in group A is lower; whereas higher incidence of temporary unilateral vocal cord palsy, permanent hypoparathyroidism, permanent unilateral vocal cord palsy, and bilateral vocal cord palsy in group A were observed. Both univariate and multivariate analyses in group A revealed that age, pathological tumor node metastasis (pTNM) stage, and histology were related to DFS (P < 0.05); while pTNM stage and histology were related to OS only in univariate analyses.Positive nodal metastases have significant prognostic value in patients with differentiated thyroid cancer in Guangdong, China and primarily reduce DFS. Moreover, patients with positive nodal metastases who are >45 years and have higher pTNM stage or follicular histology tend to have poor prognosis. Selective lymph node dissection with appropriate postoperative treatment and frequent follow-up should be accorded to these vulnerable groups of patients.

Efficacy and prognosis of neoadjuvant chemotherapy is correlated with breast cancer molecular classification.

Breast cancer is one of the most common cancers affecting women globally. Recent studies have begun to investigate the possibility of customized treatment options for individuals based on the specific cancer type. Here, we sought to analyze the relationship between the molecular classification of breast cancer and the efficacy and prognosis of neoadjuvant chemotherapy (NCT). The study included 100 breast cancer patients treated with an NCT regimen of epirubicin and docetaxel (ET) who were divided into groups based on cancer subtype (luminal, HER2 over-expression, and basal-like subtype). The nuclear classification, number of NCT cycles, pathological remission rate, and clinical curative effect, as well as the disease-free survival time (DFS) and the overall survival (OS), were compared across groups. The nuclear grade of participants in the basal-like group was significantly higher than those in the other groups but this group had fewer preoperative NCT cycles and lower pathological remission and clinical efficacy (Z=53.245, 50.077, 62.467, χ2=16.082, p<0.05). The OS and DFS of participants in the luminal subtype group were significantly higher than those in other groups while those in the basal-like subtype group were the lowest. The OS and DFS of participants who achieved pathological complete remission (pCR) through NCT treatment were significantly higher than those of the patients who had not achieved pCR through NCT treatment (χ2=9.558, 10.139, p<0.05). Therefore, we conclude that when NCT (ET regimen) is used in the treatment of breast cancer, the curative effects and prognosis appear to be correlated with the molecular classification of the tumor. Based on these results, clinicians should consider the molecular classification of the individual tumor to design the most effective treatment option.

Synthesis and In-Vitro Cytotoxicity of (E)-N,2,3-Triarylacrylamide Derivatives as Analogs of Combretastatin A-4.

A new series of (E)-N,2,3-triarylacrylamide derivatives were designed and synthesized as potent anticancer agents. Cytotoxicity of the 26 target compounds was evaluated in vitro against six cancer cell lines (HCT116, A549, MDA-MB-468, HepG2, SKNMC and SK-OV-3) by Sulforhodamine B colorimetric assay. The most promising compound, 4h, was as potent as the reference drug cisplatin (DDP). Preliminary structure-activity relationship (SAR) data provided guidance for further design and discovery of (E)- N,2,3-triarylacrylamide scaffold anticancer agents.

[Expression, purification and application of EsxB protein in Staphylococcus aureus].

OBJECTIVE: This study aimed to establish the method of expression and purification of EsxB protein, explore the EsxB antibody-positive Staphylococcus aureus (S. aureus) clinical infection status and relevance of drug resistance. METHODS: Constructed EsxB prokaryotic expression system by homologous recombination, Ni(2+) column was used to purify EsxB protein; and then ELISA was used to detect the anti-EsxB antibodies in serum of 78 patients with S. aureus infection; antimicrobial susceptibility of related S. aureus strains by automatic bacterial identification analyzer. RESULTS: EsxB prokaryotic protein expression system was constructed and EsxB protein was purified successfully; anti-EsxB antibodies were present in the serum of patients with S. aureus infection up to 28.21% (22/78). The proportion of multi-drug resistant and Methicillin-resistant S. aureus strains isolated from anti-EsxB antibodies positive patients were 100.0% (22/22), 77.3% (17/22), respectively, which were statistically higher than those strains isolated from anti-EsxB antibody-negative patients (35.7% (20/56) and 21.4% (12/56), respectively) (all P values < 0.01). CONCLUSION: Method for expression and purification of EsxB protein was established. All the S. aureus strains isolated from EsxB antibody-positive patients were multidrug resistant strains and most of them were resistant to methicillin.

[Detection and significance of transcription factors and cytokines of Th17/Treg cells in peripheral blood in the gastric cancer patients].

OBJECTIVE: To detect the expression levels of transcription factors and associated cytokines of Th17 and Treg cells in peripheral blood mononuclear cells (PBMC) of patients with gastric cancer, and explore the possible pathological mechanism of these cells involved in the development of gastric cancer. METHODS: The mRNA levels of RORgammat, FoxP3 in PBMC were determined by quantitative real-time PCR (QRT-PCR) from 57 patients with gastric cancer, 31 patients with benign gastric illness and 40 healthy people. The concentration of IL-17, IL-23, TGF-beta, IL-10 in plasma were detected by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with healthy volunteers, patients with gastric cancer showed higher levels of RORgammat and FoxP3 in PBMC (P < 0.05). The ratio of FoxP3/RORgammat in gastric cancer group was higher than that in the volunteer group and benign gastric illness group (P < 0.05). The ratio of FoxP3/RORgammat was higher in advanced disease than early disease (P < 0.05). The expressions of IL-17, IL-23, TGF-beta and IL-10 were higher in patients with gastric cancer than that in healthy volunteers (P < 0.05). In addition, The expression of TGF-beta and IL-10 were significantly increased in the advanced disease group than that in the early group (P < 0.05), but IL-17 and IL-23 was not significantly changed between the two groups (P > 0.05). CONCLUSION: There are higher levels of Th17 and Treg cells in gastric cancer patients, and it also shows a persistent predominant tendency of Treg cells and a reduced tendency of Th17 cells in advanced disease. Detecting the expression of Th17/Treg transcription factor and related cytokines would contribute to the diagnosis and prediction of the disease development and prognosis.

[Cloning and expression of human Runx3 gene obtained from human peripheral blood CD8(+) T lymphocyte].

AIM: Runx3, a type of Runt family member, plays an important role in immune regulation. In this study, we cloned and analyzed the cDNA encoding human Runx3 from T lymphocyte, expressed Runx3 protein in E.coli system, and studied the relation between Runx3 and some immune disorders or tumors. METHODS: The CD8(+) T were isolated from human peripheral blood with MACS, Runx3 cDNA was amplified by RT-PCR and cloned into pMD19-T vector, and recombinant was transformed into competent cells DH5alpha and recombinant sequencing were performed. The identical was subcloned into pQE30 vector and expressed in E.coli M15. The fusion protein was identified by Western blot. RESULTS: The 1,248 bp fragment amplified by RT-PCR was the same as the anticipated one in size and encodes 415 amino acids. Runx3 protein was gained. CONCLUSION: Human Runx3 gene was cloned and expressed in E.coli system successfully, which brought a foundation for further research on its biological function.

[Chinese traditional medicine yi kang ling to treat immunity infertility caused by anti-sperm antibody: the experimental research II].

OBJECTIVE: To observe the effects of Chinese traditional medicine Yi Kang Ling (YKL) on immunity infertility caused by anti-sperm antibodies (AsAb). METHODS: With the AsAb infertile rabbit as the experimental model, seventy-five pairs of New zealand rabbits were divided into three group including YKL treatment group (sub-divided into mini-, midi- and maxi-dosage groups), prednisone treatment group and non-treatment group. Rabbits of the YKL treatment sub-groups were further divided into M+F- (male positive, female negative), M-F+, (male negative, female positive) and M+F+ (male positive, female positive) pairs according to their initial AsAb detection results. The control group consisted of five pairs of normal rabbits. When the expected AsAb reversing ratio was achieved, the rabbits were matted and observed for the number of the pregnant and the weight of the newborn. RESULTS: Statistics showed that in M+F- pairs both the midi-dosage of YKL and prednisone treatment groups had fertility, in the mini- and maxi-dosage of YKL treatment groups, 20% of the female rabbits failed to be pregnant, while in the non-treatment group, 60% female rabbits remained sterile. The sterile ratios of the M-F+ pairs in the mini-, midi- and maxi-dosage of YKL and prednisone treatment groups were 0, 20%, 25% and 25%, respectively, while the sterile ratio in the non-treatment M-F+ group was 40%. In M+F+ groups, the sterile ratios of the three YKL sub-groups, prednisone treatment and non-treatment groups were 20%, 20% and 60% respectively. In the control group the sterile ratio was 20%. The weight of the newborn rabbits were around 50 grams with no visible malformation. CONCLUSIONS: YKL can effectively reverse the AsAb positive results, and restore the fertility of female rabbits. Mini-dosage of YKL for 45 days produced the best results and maxi-dosage of YKL had no negative effects on the weight of the newborn rabbit.