Circular RNAs (circRNAs) are endogenous non-coding RNAs with covalently closed structures, which have important functions in plants. However, their biogenesis, degradation, and function upon treatment with gibberellins (GAs) and auxins (1-naphthaleneacetic acid, NAA) remain unknown. Here, we systematically identified and characterized the expression patterns, evolutionary conservation, genomic features, and internal structures of circRNAs using RNase R-treated libraries from moso bamboo (Phyllostachys edulis) seedlings. Moreover, we investigated the biogenesis of circRNAs dependent on both cis- and trans-regulation. We explored the function of circRNAs, including their roles in regulating microRNA (miRNA)-related genes and modulating the alternative splicing of their linear counterparts. Importantly, we developed a customized degradome sequencing approach to detect miRNA-mediated cleavage of circRNAs. Finally, we presented a comprehensive view of the participation of circRNAs in the regulation of hormone metabolism upon treatment of bamboo seedlings with GA and NAA. Collectively, our study provides insights into the biogenesis, function, and miRNA-mediated degradation of circRNAs in moso bamboo.
RNA, Circular/metabolism* ; Multiomics ; Poaceae/metabolism* ; Seedlings/genetics* ; Hormones/metabolism* ; MicroRNAs/metabolism* ; Gene Expression Regulation, Plant

ObjectiveTo investigate the clinical features, diagnosis, treatment, and prognosis of autoimmune pancreatitis (AIP) alone versus AIP with IgG4 sclerosing cholangitis (IgG4-SC). MethodsA retrospective analysis was performed for the clinical data of 40 patients with type 1 AIP who were admitted to The First Affiliated Hospital of Zhengzhou University from June 2015 to January 2020, among whom 29 patients had AIP alone and 11 had AIP with IgG4-SC. The two groups were compared in terms of clinical manifestations, laboratory examination, imaging findings, treatment, and prognosis. The t-test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the Fisher’s exact test was used for comparison of categorical data between two groups. The Kaplan-Meier method was used to calculate recurrence rate and plot recurrence curve, and the log-rank test was used for univariate analysis. ResultsCompared with the AIP group, the AIP+IgG4-SC group had significantly higher number of affected organs ［3.0(3.0-4.0) vs 3.0(1.5-3.5), Z=-2.172, P=0035］ and response index before treatment ［12.0(12.0-15.0) vs 12.0(9.0-13.5), Z=-2.157, P=0.032］. The AIP+IgG4-SC group had a significantly higher median serum IgG level than the AIP group ［21.0(15.8-23.7) g/L vs 14.8(13.3-15.7) g/L, Z=-2.711, P=0.004］. During the median follow-up time of 15.8 (6.5-31.3) months, the AIP+IgG4-SC group had a significantly higher recurrence rate than the AIP group (χ2=8.155, P=0.004). ConclusionPatients with AIP and IgG4-SC tend to have higher serum IgG4 level, number of affected organs, and recurrence rate than those with AIP alone. Early identification, diagnosis, and treatment can reduce the recurrence rate of AIP.

Objective: To explore the clinicopathological characteristics of primary hepatic neuroendocrine neoplasm (HNEN) and metastatic HNEN from digestive tract, to screen the risk factors of hepatic metastasis of gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN) and to analyze the differences between primary and metastatic HNEN in clinical features, diagnosis, treatment and prognosis. Methods: From January 2010 to June 2017, the clinical data of 182 patients with HNEN admitted at The First Affiliated Hospital of Zhengzhou University were retrospectively analyzed, including 39 cases of primary HNEN, 129 cases of metastatic HNEN and 14 cases of HNEN with unknown primary lesions. Chi-square test and t test were performed to analyze the pathologic characteristics among groups. Logistic regression method was used to analyze the risk factors of hepatic metastasis. Kaplan-Meier method and log-rank test were used for survival analysis. Cox model was used for the prognostic multivariate survival analysis. Results: Metastatic HNEN from digestive tract was more common in male (70.5%, 91/129). The case number of serological tumor biomarkers neuronspecific enolase and alpha-fetoprotein positive in primary HNEN were two cases and one case, respectively, and the positive rates in metastatic HNEN were 37.2% (32/86) and 6.4% (7/110). Most primary HNEN was single lesion (61.5%, 24/39), while multiple lesions were more common in metastatic HNEN (78.3%, 90/115). Primary HNEN mainly occurred in the right lobe of the liver (44.7%, 17/38), while metastatic HNEN located simultaneously in the left and right lobes of the liver (68.4%, 78/114). There were significant differences between primary HNEN and metastatic HNEN in tumor number, pathological grading, location of tumors and maximum diameter of tumors (χ²=21.264, 11.696, 19.461 and 4.547, all P<0.05). The median survival time of patients with primary HNEN and metastatic HNEN were 17.0 months and 10.0 months, and there was a significant difference in survival curves between the two groups (χ²=7.235, P=0.007). The type of hepatic tumors (primary or metastatic)(P=0.002), pathological grading of hepatic tumors (P=0.044), lymph node metastasis (P=0.024), the growth pattern of tumors (P<0.01) and treatment methods (P=0.018) were the independent factors for the prognosis of patients. Conclusions There are significant differences between primary HNEN and metastatic HNEN in tumor number, size and location. The type of hepatic tumors, pathological grading, lymph node metastasis, growth pattern of tumors and treatment methods are the independent factors for the prognosis of patients. Early topical treatment and combination treatment can help to prolong survival time of HNEN patients.

Objective To explore the clinicopathologic characteristics,to screen risk factors of metastasis and to analyze the diagnosis,treatment and prognosis of gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN).Methods From January 2010 to November 2015,the clinical data of 405 patients with GEP-NEN were retrospectively analyzed.GEP-NEN tumors were classified as neuroendocrine tumor (NET,G1 and G2 grade),neuroendocrine carcinoma (NEC,G3 grade) and mixed adenoendocrine carcinoma (MANEC,G3 grade).The clinicopathologic characteristics were summarized.The staining characteristics of synaptophysin (Syn),chromogranin A (CgA) and CD56 of tumor tissues were analyzed by immunohistochemistry.x2 and t test were performed to analyze differences in pathologic characteristics between groups.Logistics regression method was used to analyze the risk factors of metastasis.KaplanMeier method and Log-rank test were used for survival analysis.Results The mean age of patients with GEP-NEN was (54.7± 13.3) years.Gastric NEN was the most common GEP-NEN (98 cases,24.2％),followed by 95 cases (23.5％) with NEN in rectum,86 cases (21.2％) in pancreas and 50 cases (12.3％)in esophagus.Among them,47 cases (11.6％) were functional GEP NEN and 358 cases (88.4％) were non-functional GEP-NEN.According to pathologic diagnosis,227 cases (56.0％) were NET,125 cases (30.9％) were NEC and 16 cases (4.0％) were MANEC.According to tumor classification,120 cases (29.6％) were grade G1,108 cases (26.7％) were grade G2 and 177 cases (43.7％) were grade G3.Immunohistochemistry staining positive ratesof Syn,CgA and CD56 were 97.4 ％ (381/391),44.0 ％ (121/275) and 83.9％(291/347),respectively.The median (lower quartile,upper quartile) diameter of grade G1,G2 and G3 tumors were 1.0 cm (0.6 cm,1.5 cm),1.5 cm (1.0 cm,2.5 cm),4.0 cm(2.5 cm,6.0 cm),respectively,and the difference was statistically significant (Z =99.171,P ＜ 0.01).The positive rate of CgA of grade G3 tumor was lower than that of grade G1 and G2(x2 =7.078 and 11.391,both P＜ 0.01).The results of Logistic regression analysis revealed that tumor size and pathologic classification were the important predictors of metastasis.The median survival time of metastasis group and non-metastasis group of grade G3 was 12.0 months and 41.5 months,and there was a significant difference between the two groups by Log-rank test (x2 =37.075,P＜0.01).Conclusions GEP-NEN may occur at any part of the digestive system.There are differences in tumor size positive rate of,immunohistochemistry staining and the primary site of tumors with different pathological grading.The tumor diameter and pathologic classification are the important predictors of metastasis.The prognosis of metastasis group is worse than that of non-metastasis group.

Objective To evaluate the effect of anti-RANTES monoclonal antibody in combination with ciclosporin (CsA)upon inhibiting the rejection response during secondary heart transplantation in mouse models. Methods BALB/c mouse models were used as the donors and C57BL/6 mice were utilized to establish secondary heart transplantation recipient models. The animals were randomly divided into the control (physiological saline,n =6 ),A (anti-RANTES monoclonal antibody treatment,n =6 ),B (CsA treatment,n =6 ) and C groups (anti-RANTES monoclonal antibody combined with CsA treatment,n=6). The survival time of heart after secondary transplantation was observed. The degree of acute heart rejection was assessed by histopathological analysis. The relative expression levels of RANTES,interleukin(IL)-2,IL-1 0,interferon(IFN)-γand transcription growth factor(TGF)-βmessenger ribonucleic acid (mRNA)in the heart grafts were quantitatively measured by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The serum levels of RANTES,IFN-γ,IL-2,IL-1 0 and TGF-βwere detected by enzyme-linked immune absorbent assay (ELISA). Results The heart grafts of all mice survived after secondary cardiac transplantation. Compared with the control group,the survival time of hearts in group A,B and C was significantly prolonged (all P<0. 01 ). Pathological staining revealed that the quantity of infiltrated inflammatory cells in group C was significantly decreased than those in the other groups. The expression levels of heart RANTES,IFN-γand IL-2 mRNA in group C were significantly down-regulated, whereas the expression levels of IL-1 0 and TGF-βmRNA were considerably up-regulated compared with those in the other three groups (all P<0. 05). The serum levels of RANTES,IL-2 and IFN-γin group C were significantly down-regulated, whereas the serum contents of IL-1 0 and TGF-βwere considerably up-regulated compared with those in the other three groups (all P<0. 05 ). Conclusions Combined application of anti-RANTES monoclonal antibody and CsA can effectively induce the immune tolerance to secondary cardiac transplantation and prolong the survival time of the cardiac grafts in mouse models.

Objective To study the influence of Apo E gene knockout on the lymphocytes . Methods Cells from the peripheral blood, spleen and bone marrow of Apo E knockout and wildtype mice were stained with kinds of antibodies , and analyzed by flow cytometry .Results Compared with wildtype mice , significant differences were found in B and T lymphoctes in the peripheral blood and spleen , but there was no significant difference in pre B cells , T lymphocytes in the thymus and long term hematopoietic stem cell in the Apo E knockout mice .Conclusion Numbers of B lymphocytes decreased in the peripheral blood and spleen , but there was no significant difference in B , T lymphocytes development , and numbers of long term hematopoietic stem cell in Apo E gene knockout mice .

Objective To study the influence of IL-33 on the Hematopoietic stem cells and progenitor cells . Methods Cells from the peripheral blood , spleen, thymus and bone marrow were stained with indicated antibodies and analyzed by flow cytometry . The LT-HSCs were sorted and culture using in vitro clonogenic assay . Results The percentage of B cells and T cells was decreased and the percentage of M cells was increased in the peripheral blood from IL -33 transgenic mice .Compared with the wildtype mice , the number of HSCs , MPPs and CLP was decreased;meanwhile the number of CMP and GMP was increased in the bone marrow from IL-33 transgenic mice .An in vitro clonogenic assay showed that LT-HSCs increased the ability to self-renew from IL-33 transgenic mice .And the percentage of S-G2-M stage hematopoietic stem cell was increased from IL-33 transgenic mice .Conclusion IL-33 increase the myeloid differentiation in hematopoietic stem cells .

Objective To investigate the influence of Siraitia Grosvenori and Rehmannia Glutinosa on the Hematopoietic stem cells proliferation and function .Methods Cells from the peripheral blood , spleen and bone marrow of mice were stained with indicated antibodies , and analyzed by flow cytometry .Mice were divided 3 groups:control group, Siraitia Grosvenori treatment group and Rehmannia Glutinosa treatment group .After 4.5Gy IR treatment, mice divided 4 groups: control group, 4.5Gy IR treatment and feed with normal food, 4.5Gy IR treatment and feed with Siraitia Grosvenori and 4.5Gy IR treatment and feed with Rehmannia Glutinosa for 1 month.Results Mice fed with Siraitia Grosvenori and Rehmannia Glutinosa decreased the percentage of B cells and increased the percentage of M cell .For HSCs, the number of HSCs was increased , especially the number of LT-HSCs.After 4.5Gy IR treatment, mice fed with Siraitia Grosvenori and Rehmannia Glutinosa increase the number of HSCs , and increased the percentage of M cells . Conclusion Siraitia Grosvenori and Rehmannia Glutinosa promote the hematopoietic stem cells and progenitor cells proliferation and function and recover the damage that caused by IR treatment .

Objective To understand the difference in cognition of nursing needs of different gender patients with hospitalized hepatitis .Methods The questionnaire of nursing needs of hospitalized hepatitis patients was self-designed through the retrieval of literature and the nursing of hepatitis in practice , and 164 patients with hospitalized hepatitis received the investigation of nursing needs and satisfaction .Results One hundred and sixty-four questionnaires were issued and 110 questionnaires were included in this study with an effective response rate of 67.1%.No differences were found in the physiology , security, emotional and attribution, self-realization in different gender patients (P>0.05).The percentage of “to do explanation before all kinds of operation, and your consent” in the items of respect in men was 56.7%(47/83), and was 25.9%(7/27) in women, and the difference was statistically significant (P=0.017), and no differences were found in the other items of respect (P>0.05).The percentage of general and great satisfaction in men was 78.3%(65/83), and was 51.9%(14/27) in women, and the difference was statistically significant (P=0.007). Conclusions The cognition of nursing needs of different gender patients with hospitalized hepatitis is almost the same, but women have a higher requirement to the nursing satisfaction .

Objective To investigate the in vivo effect of silenced actin-associated protein Transgelin on the growth of human pancreatic carcinoma xenograft in nude mice.Methods Human pancreatic cancer cell line BxPC3 were transfected with small hairpin RNA (shRNA) eukaryotic expression vector targeting Transgelin gene.RT-PCR and Western blot were used to analyze Transgelin expression after transfection.24 animal models were randomly divided into three groups with 8 in each:Experimental group (transplanted BxPC3/Transgelin shRNA),negative control group (transplanted BxPC3/Neo) and untreated group (transplanted BxPC3).Tumor size was measured weekly.All mice were sacrificed after 28 days.Tumor volume was calculated,inhibitory effect was analyzed.Immunohistochemical staining of paraffin sections for Transgelin and proliferating cell nuclear antigen (PCNA) proteins were performed.Results Tumors varied in sizes among 3 groups (all P ＜ 0.05).On day 21 and 28 tumor was significantly smaller in experimental group than those in control groups.Tumor weighed(0.74 ±0.21) g in experimental group,lower than that in negative control group(1.42 ± 0.28) g and untreated group(1.59 ± 0.24) g (all P ＜ 0.05).The inhibitory effect was 53.5％ in experimental group.The PCNA index was significantly lower in experimental group than those in control groups (all P ＜ 0.05).Conclusions Deletion of Transgelin gene can significantly inhibit the proliferation and tumor growth of BxPC3 cells in nude mice.