[Analysis of the common respiratory viruses in children with acute respiratory infection in a hospital in Lanzhou City from 2021 to 2022].

To explore the situation of 8 common respiratory pathogens in children with acute respiratory infection (ARI) from 2021 to 2022.The retrospective study selected 8 710 ARI patients from September 2021 to August 2022 in the Maternal and Child Health Hospital of Gansu Province as the study object, patients aged 0 to 17 years old, including 5 048 male children and 3 662 female children. Indirect immunofluorescence was used to detect 8 common respiratory pathogens, including influenza virus A (FluA), influenza virus B (FluB), parainfluenza virus (PIV), respiratory syncytial virus (RSV), adenovirus (ADV), Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP), and Coxsackie virus group B (CoxB) IgM antibodies. χ2 test was used to analyze the results. The results showed that 1 497 of 8 710 children with ARI were positive, with a positive rate of 17.19%. The detection rate of MP among 8 common respiratory pathogens was 11.34%, accounting for 66.0%, followed by FluB, CoxB, PIV, RSV, ADV, FluA and CP, accounting for 13.83%, 9.55%, 6.01%, 2.61%, 1.47%, 0.40% and 0.13%, respectively. Respiratory tract viruses (FluA, FluB, RSV, ADV, PIV, CoxB) accounted for 33.86%.There were significant differences in the detection rates of PIV, ADV and MP among children of different genders (χ2=6.814, 5.154 and 17.784, P<0.05). The detection rate of school-age children (6-17 years old) was the highest, accounting for 33.27% (184/553). The detection rates of 8 common respiratory pathogens in patients with ARI were higher in spring and winter and lower in summer and autumn. To sum up, from 2021 to 2022, MP and FluB infection were dominant in ARI patients in our hospital. The peak period of 8 common respiratory pathogens was in spring and winter. The physical examination rate of 8 common respiratory pathogens in ARI patients aged 6-17 years old was the highest.

[Effect of REG3A on proliferation and invasion of glioma cells by regulating PI3K/Akt signaling pathway].

Objective: To investigate the effects of regenerating islet-derived protein 3A (REG3A) on the proliferation and invasion of glioma cells and its molecular mechanism. Methods: Five low-grade, five high-grade glioma tissues and ten adjacent tissues from glioma patients who underwent surgery at Linyi People's Hospital from October 17, 2017 to October 18, 2018 were collected. Human glioma cell lines (SF295, U251, TG905, A172, CRT) and a primary glioma cell line PT-1 were cultured in vitro. The protein and mRNA expressions of REG3A in these tissues and glioma cell lines were detected by Western blot and reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). SF295 cells were infected with lentivirus and labeled as REG3A plasmid transfection group, and the TG905 cells were transfected with si-REG3A by liposome transfection reagent and labeled as si-REG3A transfection group. At the same time, the empty transfection control and blank control groups were set up. Glioma cells were treated with REG3A recombinant protein alone or in combination with Akt1/2 inhibitors. Cell counting kit-8 (CCK-8) and cell scratch assay were used to detect cell proliferation and invasion, respectively. Western blot was used to detect the protein expression of N-cadherin, vimentin and phosphorylation of Akt (p-Akt) in REG3A overexpressed and knockdown glioma cells. Results: RT-qPCR results showed that the mRNA expression levels of REG3A in glioma cells in each group were U251 (2.129±0.13), TG905 (2.22±0.59), CRT (5.02±0.31), A172 (6.62±1.34) and PT-1 (9.18±0.61), respectively, higher than its expression in SF295 cells (1.00±0.18, P<0.001). The mRNA expression level of REG3A in high-grade glioma tissue samples (3.18±2.92) was higher than that in the control group (1.00±1.14, P=0.031) and low-grade glioma group (0.90±0.67, P=0.014). The results of western blot and immunohistochemical staining were consistent with that of RT-qPCR. The migration rate of cells in si-REG3A transfection group [(60.57±5.30)%] was lower than that of the empty transfection group [(84.18±13.63)% (P=0.038)] and blank control group [(79.65±12.09)% (P=0.076)]. The results of the scratch experiment showed that the migration rate of cells in REG3A plasmid transfected cells in the SF295 group was (96.05±6.41)%, which was significantly higher than that of empty transfected cells [(74.47±8.23)%, P=0.021)]. REG3A recombinant protein could up-regulate the expression of N-cadherin, vimentin and p-Akt in SF295 cells. Compared with the control group [(100.00±2.53)%], the proliferation rate in the REG3A recombinant protein group [(117.70±10.24)%] was significantly up-regulated, and the proliferation rate in the REG3A recombinant protein+ Akt inhibitor group [(98.31±3.64)%] was significantly lower than that of the REG3A recombinant protein group (P=0.017). The migration rate of the REG3A recombinant protein+ Akt inhibitor group was (63.35±4.06)%, which was significantly lower than (89.26±11.07)% of the REG3A recombinant protein group (P=0.019). Conclusion: REG3A can promote the proliferation and invasion of human glioma cells by activating the PI3K/Akt signaling pathway.

[A clinicopathological classification of space-occupying lesions of the orbit in 1 913 patients from 2000 to 2021].

Objective: To investigate the histopathological classification of orbital space-occupying lesions. Methods: This is a retrospective case series study. The clinical and pathological data of 1 913 tissue specimens from 1 913 patients with space-occupying lesions of the orbit which were examined in the Second Affiliated Hospital, Zhejiang University School of Medicine from January 2000 to December 2021 were collected. The mass lesions were classified based on histogenesis, pathological nature and age. Results: There were 913 males (47.7%) and 1 000 females (52.3%). The lesions were benign in 1 489 patients (77.8%) and malignant in 424 patients (22.2%). Based on histogenesis, there were 521 vasculogenic lesions (27.2%), which rancked first, 407 cystoid lesions (21.3%), 277 lymphoproliferative lesions (14.5%), 182 lacrimal gland lesions (9.5%) and 121 inflammatory lesions (6.3%). By pathological nature, there were 1 489 benign lesions, including cavernous hemangioma (275, 14.4%), dermoid cyst (225, 11.8%), other hemangiomas (199, 10.4%), epidermoid cyst (136, 7.1%) and benign mixed tumor of the lacrimal gland (134, 7.0%), and 257 malignant lesions, including lymphoma (210, 11.0%) and sebaceous gland carcinoma (47, 2.5%). The age of all patients ranged from 0 to 90 years, while 247 lesions (12.9%) occurred in patients aged 0 to18 years, 1 270 lesions (66.4%) in patients aged 19 to 59 years, and 396 lesions (20.7%) in patients aged 60 to 90 years. Conclusions: In 22 years, almost 2/3 benign orbital lesions in the Second Affiliated Hospital, Zhejiang University School of Medicine occurred in young and middle-aged patients, and males were fewer than females. The most common benign orbital tumors was cavernous hemangioma, followed by dermoid cyst and epidermoid cyst. And the most common malignant orbital tumor was lymphoma, which occurred more frequently in older patients.

[Cognition and related factors on the use of HIV non-occupational post-exposure prevention among men who have sex with men].

Objective: To understand the cognition and related factors on the use of HIV non-occupational post-exposure prophylaxis (nPEP) among men who have sex with men (MSM). Methods: The snowballing method was applied to recruit research subjects who were ≥18 years old, had sex with men in the past three months, and were aware of nPEP in MSM groups in Beijing, Shenzhen, and Kunming from March 15 to April 14, 2019. Data on social demographics, behavioral characteristics, basic knowledge of nPEP, consultation, and using nPEP were collected through "i guardian Platform". The logistic regression model was used to analyze the related factors affecting the use of nPEP. Results: Among 1 809 investigated, 39.8% (720 persons) were aware of the basic knowledge of nPEP, 33.4% (605 persons) had consulted nPEP, and 15.0% (271 persons) had used nPEP. In addition, multivariate logistic regression analysis showed that factors as whether to have sex with men infected with HIV in the last three months (OR=2.58, 95%CI: 1.64-4.07), the frequency of HIV testing in the past year (OR=2.47, 95%CI: 1.28-5.11), nPEP knowledge awareness (OR=0.70, 95%CI: 0.49-0.99), whether to consult nPEP (OR=70.98, 95%CI: 40.51-136.83) were related to the use of nPEP. Conclusions: MSM still have poor cognition of nPEP. It is necessary to strengthen the publicity and education of nPEP in MSM and promote the use of nPEP after HIV exposure as soon as possible.

[CRISPR/Cas9-mediated microRNA-21 knockout increased imatinib sensitivity in chronic myeloid leukemia cells].

Objective: To observe the effects of miR-21 knockout on proliferation and drug resistance in K562/G01 cells, and to preliminarily explore the mechanism of imatinib sensitivity by knocking out miR-21 in K562/G01 cells. Methods: Using CRISPR/Cas9 to knock out the miR-21 gene in K562/G01 cells, and single-cell-derived clones of miR-21 knockout were obtained by genomic DNA PCR screening, Sanger sequencing, and real-time PCR. We used MTT and cell colony formation assays to assess the cell proliferation, and determined imatinib sensitivity by MTT assay and Annexin-Ⅴ-APC/7-AAD double staining flow cytometry. Using western blot, we examined the potential mechanisms affecting imatinib sensitivity by knocking out miR-21 in K562/G01 cells. Results: Three miR-21 knockout K562/G01 single-cell-derived clones were successfully constructed. The mutation efficiency mediated by CRISPR/Cas9 was 7.12%-8.11%. MiR-21 knockout inhibited the proliferation of K562/G01 cells; the clone formation rates of WT and 1#, 2#, 6# K562/G01 single-cell clones were (57.67±8.25) %, (26.94± 5.36) %, (7.17±2.11) %, (31.50±3.65) %, respectively. MiR-21 knockout increased the sensitivity of K562/G01 cells to imatinib, IC(50) of imatinib in WT, and 1#, 2#, 6# K562/G01 single-cell clones were (21.92±1.36) µmol/ml, (3.98±0.39) µmol/ml, (5.38±1.01) µmol/ml, (9.24±1.36) µmol/ml. After the knockout of miR-21, the activation of PI3K/Akt signaling molecules was inhibited, while the expression of P210(B)CR-ABL and p-P210(BCR-ABL) was downregulated; however, the expression of PTEN was not affected. Conclusion: The knockout of miR-21 can suppress cell proliferation and improve sensitivity to imatinib in K562/G01 cells, which may be achieved by inhibiting the PI3K/AKT signaling pathway and BCR-ABL expression.

[An interpretation of global consensus on the diagnosis and management of limbal stem cell deficiency].

Limbal stem cell deficiency is an ocular surface disease with the imbalance of corneal epithelial homeostasis caused by decrease of number or weakening of function of limbal stem cells. In 2019 and 2020, the international limbal stem cell deficiency working group developed and released the international consensus on the definition, classification, diagnosis, staging and management of limbal stem cell deficiency, which provided a standard protocol for related research and clinical diagnosis and treatment of limbal stem cell deficiency. In order to help Chinese ophthalmologists to properly understand its core components, there is a need to make in-depth interpretation to the key and difficult contents of the international consensus. (Chin J Ophthalmol, 2021, 57: 95-99).

[Research progress on limbal stem cell transplantation].

Limbal Stem Cell Deficiency (LSCD) is an ocular surface disease caused by the decrease of the quantity and dysfunction of limbal stem cell, which is characterized by conjunctivalization and other signs of epithelial dysfunction. For sever LSCD, surgery is the main treatment way. Recently, plenty of researches published the outcomes of different operation methods. This article summarized five major operations, including conjunctival limbal autograft (CLAU), simple limbal epithelial transplantation (SLET), limbal allograft, cultivated limbal stem cell transplantation (CLET) and cultivated oral mucosal epithelial transplantation (COMET). (Chin J Ophthalmol, 2020, 56:956-960).

Long non-coding RNA LINC00173 serves as sponge for miR-338-3p to promote prostate cancer progression via regulating Rab25.

OBJECTIVE: Long non-coding RNA LINC00173 (LINC00173) has been shown to facilitate the progression of a number of malignancies. In this study, we aimed to investigate the function of LINC00173 on prostate cancer (PCa) and discover the potential regulatory mechanism. PATIENTS AND METHODS: RT-PCR was used to determine the levels of LINC00173, miR-338-3p and Rab25 in PCa patients and cell lines. The clinical significance of LINC00173 was statistically analyzed in 124 PCa patients. CCK-8, colony formation, transwell, scratch wound, Ethynyldeoxyuridine (EdU) assays and flow cytometry assays were used to detect the proliferation, apoptosis, invasion and migration of PCa cells. The mechanism of LINC00173 action was explored through bioinformatics, RNA pull-down assays and Luciferase reporter assays. RESULTS: We observed that the expression of LINC00173 and Rab25 was distinctly upregulated in both PCa specimens and cell lines, while miR-338-3p expression was significantly down-regulated. High LINC00173 expression was associated with Gleason score, preoperative PSA level and reduced patient survivals. Functional assays revealed that knockdown of LINC00173 suppressed the proliferation, migration and invasion of PCa cells, and promoted apoptosis. Mechanistically, LINC00173 acted as a competitive endogenous RNA in PCa and increased Rab25 expressions via sponging miR-338-3p. Moreover, LINC00173 promoted PCa progression by interacting with miR-338-3p and Rab25. CONCLUSIONS: Our findings, for the first time, identified a novel PCa-related lncRNA, LINC00173 which might serve as an oncogene in PCa. The discovery of the LINC00173/miR-338-3p/Rab25 pathways provided new thinking for the treatments of PCa.

[Effect of circular RNA hsa_circ_0008898 on oral squamous cell carcinoma and its mechanism].

Objective: To investigate the effect and molecular mechanism of hsa_circ_0008898 on the cell proliferation, migration, invasion and tumor formation of oral squamous cell carcinoma (OSCC). Methods: Quantitative real-time PCR (qPCR) and Western blotting were used to detect the expression of hsa_circ_0008898, miR-197-5p and ras homolog gene family member A (RHOA) in OSCC tissues, adjacent tissues, OSCC cells and human normal oral keratinocytes (NOK). CAL27 and SCC-25 cells were transfected with si-hsa_circ_0008898#1 (knockdown group 1), si-hsa_circ_0008898#2 (knockdown group 2), hsa_circ_0008898 (circ overexpression group) and blank plasmid (circ blank group), respectively. Then miR-197-5p inhibitor (inhibition group) and blank plasmid (inhibition control group) were transfected into hsa_circ_0008898 knockdown cells (knockdown group 1). CAL27 and SCC-25 cells were transfected with miR-197-5p mimics (miR overexpression group) and blank plasmid (miR blank group), and then transfected hsa_circ_0008898 vector (co-transfection group 1), RHOA vector (co-transfection group 2) and blank plasmid (co-transfection control group) in cells overexpressing miR-197-5p. Cell counting kit 8 (CCK-8), colony formation, Transwell and scratch test were used to detect cell proliferation, cloning ability, cell cycle distribution, cell invasion and migration ability. Ten nude mice were equally divided into two groups, with 5 mice in each group. SCC-25 cells transfected with blank plasmid (control group) and SCC-25 cells transfected with sh-hsa_circ_0008898 (knockout group) were subcutaneously injected into the armpit. The volume and mass of the tumor were measured. Results: The expressions of hsa_circ_0008898 (2.89±0.72) and RHOA (2.62±0.21) in OSCC tissues were significantly higher than those in para-carcinoma tissues (1.00±0.48, 1.00±0.11, respectively), while the expression of miR-197-5p in OSCC tissues （0.46±0.24） was significantly lower than that in para-carcinoma tissues (1.00±0.42) (P<0.05). Compared with NOK, the expression of hsa_circ_0008898 and RHOA in CAL27 and SCC-25 cells increased significantly, while the expression of miR-197-5p decreased (P<0.05). Compared with circ blank group, the cell viability, colony formation, scratch healing rate and invasive cell number of CAL27 and SCC-25 cells in knockdown group 1 and group 2 were significantly decreased, while the proportion of cells in G1 phase was significantly increased (P<0.05). Compared with inhibition control group, the cell viability, colony formation, scratch healing rate and invasive cell number of CAL27 and SCC-25 in inhibition group were significantly increased, while the proportion of cells in G1 phase was significantly decreased in inhibition group (P<0.05). Compared with miR blank group, the cell viability, colony formation, scratch healing rate and invasive cell number of CAL27 and SCC-25 in miR overexpression group were significantly decreased, while the proportion of cells in G1 phase was significantly increased in miR overexpression group (P<0.05). Compared with co-transfection control group, the cell viability, colony formation, migration area and invasive cell number of CAL27 and SCC-25 in co-transfection group2 were significantly increased, while the proportion of cells in G1 phase was significantly decreased in co-transfection group 2 (P<0.05). The volume and mass of transplanted tumor in knockout group ï¼»(660.4±67.8) mm(3 )and (0.60±0.06) g, respectivelyï¼½ were significantly lower than those in control group ï¼»(1 210.4±198.9) mm(3) and (1.00±0.12) g, respectivelyï¼½. Conclusions: Knockdown of hsa_circ_0008898 inhibited OSCC cells proliferation, cloning, migration and invasion and induced cell cycle arrest in vitro by regulating the miR-197-5p/RHOA. Additionally, Knockdown of hsa_circ_0008898 also inhibited tumor formation of OSCC cells in vivo.

[The 474th case: anemia, ostealgia, proteinuria].

A 49-year-old woman was admitted to hospital with intermittent dizziness and fatigue for 7 years. The symptoms were aggravated and accompanied by bone pain for more than 4 months. She was referred to our hospital. Laboratory tests and imaging findings suggested that acquired Fanconi Syndrome (FS) was associated with smoldering multiple myeloma (MM). Renal biopsy and electron microscopy confirmed the diagnosis of proximal light chain tubular disease (LCPT). LCPT causes proximal tubular dysfunction, which is characterized by the cytoplasmic crystal deposition usually kappa monoclonal light chain in the proximal tubule. MM with FS and LCPT is less common in clinical practice because it is difficult to diagnose. This is a typical case focusing on the differential diagnosis of monoclonal gammopathy of renal significance(MGRS) such as LCPT and plasma cells diseases.

[Alveolar soft part sarcoma in children: a clinicopathological study of 13 cases].

Objective: To investigate the clinicopathological manifestations, molecular genetic, diagnostic histology and differential diagnosis of alveolar soft part sarcoma (ASPS) in children. Methods: A total of 13 cases of ASPS diagnosed at Beijing Children's Hospital from August 2009 to November 2018 were collected. HE staining, histochemical staining for PAS and D-PAS, immunohistochemical (IHC) staining for TFE3, INI1 and CD68 and florescence in situ hybridization (FISH) for TFE3 gene translocation were performed. Results: There were four males and nine females, age ranged from 1 year and 2 months to 13 years and 8 months (mean 7.8 years); and four patients were under 5 years old. Histologically, the tumors showed a distinctive and characteristic nested or organoid growth pattern (11 cases) or solid, diffuse growth (2 cases). The tumor cells possessed abundant eosinophilic, or glycogen-rich and clear to vacuolated cytoplasm. The chromatin was relatively dispersed, with prominent and pleomorphic nucleoli; mitotic figures were rare. Vascular invasion was frequently seen. IHC staining showed specific nuclear TFE3 staining. The tumor cells were also positive for INI1,CD68 and vimentin; but were negative for MyoD1, Myogenin, CK and S-100 protein. Seven cases showed PAS and D-PAS staining, with fuchsia acicular or rod-shaped crystals in tumor cytoplasm. Nine cases showed TFE3 break-apart signals by FISH. Conclusions: ASPS is a rare soft tissue sarcoma in children. Compared with ASPA in adults, it has both similarities and unique clinicopathologic characteristics. The diagnosis needs to be confirmed by combining clinical, pathologic, IHC and genetic testing.

[Clinical effect of nutritional and psychological intervention combined with pulmonary rehabilitation exercise on patients with chronic obstructive pulmonary disease].

Objective: To explore the clinical effect of nutritional and psychological intervention combined with pulmonary rehabilitation exercise on patients with chronic obstructive pulmonary disease (COPD). Methods: A total of 260 patients with COPD admitted to the Sixth Affiliated Hospital of Kunming Medical University from October 2014 to October 2017 were included. They were divided into mild, moderate, severe and extremely severe groups according to forced expiratory volume in one second predicted (FEV(1)%prep) of pulmonary function. The patients were divided into control group and comprehensive management group according to the random number table method. The control group was given routine treatment including smoking quitting persuasion, vaccination, oxygen therapy and standardized medication. The comprehensive management group was given additional nutritional support, psychological intervention and pulmonary rehabilitation exercise. The data of the lung function indexes (FEV(1)%prep, FEV(1)/FVC, PaO(2), PaCO(2)), nutritional indexes [body mass index (BMI), albumin (ALB), nutrition risk screening (NRS)2002], anxiety and depression scores, 6-minute walking distance (6MWD), modified medical research council (mMRC) dyspnea scale, COPD assessment test (CAT), St. George's score, and frequency of acute exacerbations were compared between two groups after 12 months of treatment. Results: After 12 months' treatment, PaO(2) in the comprehensive management group was significantly higher than that in the control group [(51.1±7.2) vs (47.0±9.1) mmHg] (1 mmHg=0.133 kPa); Nutritional risk (NRS2002) decreased obviously [(1.1±1.1) vs (2.2±1.0)]; anxiety score [(4.1±2.2) vs (5.6±2.7)]; depression score [(4.1±2.0) vs (5.5±2.6)] and St. George's score [(36.8±20.8) vs (48.6±19.5)] decreased significantly (P<0.05). And the 6MWD was significantly farther [(368.4±72.0) vs (343.4±75.0) m] in management group. The frequency of acute exacerbations was significantly reduced in the mild, moderate and severe groups (P<0.05). But there was no significant difference in FEV(1)%prep, FEV(1)/FVC, PaCO(2), BMI, ALB, mMRC score and CAT score. Conclusion: Nutritional and psychological intervention combined with pulmonary rehabilitation exercise can reduce the nutritional risk and the frequency of acute exacerbations in patients with COPD, relieve anxiety and depression state and improve the quality of life.

[A clinicopathological classification analysis of ocular mass lesions in 7 910 cases].

Objective: To investigate the anatomical region, histopathological classification and histogensis distribution of ocular mass lesions in South China. Methods: Retrospective cases study. The clinical and pathological data of 7 910 samples with ocular (adnexal) tumors or proliferative lesions which were examined from January 2000 to May 2018 were retrospectively retrieved. The constituent ratios of ocular mass lesions in different anatomical regions and histogenesis have been analyzed. Results: There were 3 445 males and 4 465 females aged from 3 months to 106 years. Classification by anatomical region. Eyelid 4 976 cases (62.9%): benign-pigmented nevus (31.7%, 1 342/4 235), squamous cell papilloma (12.3%, 519/4 235), seborrheic keratosis (9.4%, 396/4 235); malignant-basal cell carcinoma (48.5%, 359/741), sebaceous gland carcinoma (34.4%, 255/741), squamous cell carcinoma (12.3%, 91/741). Ocular surface 1 449 cases (18.3%): benign-pigmented nevus (26.6%, 359/1 348), squamous cell papilloma (12.8%, 173/1 348); malignant-lymphoma (34.7%, 35/101), squamous cell carcinoma (30.7%, 31/101).Orbit 1 485 cases (18.8%): benign-hemangioma (28.5%, 332/1 167), lacrimal gland (duct) cyst(13.2%, 154/1 167); malignant-lymphoma (44.7%, 142/318), adenoid cystic carcinoma (10.1%, 32/318). Classification by histogenesis: epithelial 2 145 cases (27.1%), cutaneous appendages 378 cases (4.8%), cystoid 1 068 cases (13.5%), mesenchymal 748 cases (9.5%), lymph-hematopoietic 225 cases (2.8%), neurogenic 31 cases (0.4%), melanocytic 1 765 cases (22.3%), others 1 550 cases (19.6%). Conclusions: Over the past 18 years, the ocular tumors identified at the Second Affiliated Hospital, Zhejiang University School of Medicine most frequently occur in eyelid and originate from epithelium. The most common types are as followings. Benign lesions: pigmented nevus, squamous cell papilloma are the most common types for eyelid and ocular surface, whereas hemangioma, lacrimal gland (duct) cyst and epidermoid cyst are the most common types for orbit. Malignant cancers: basal cell carcinoma is the most prevalent disease in eyelid, whereas lymphoma occurs more frequently in ocular surface and orbit. (Chin J Ophthalmol, 2019, 55: 847-853).

Hsa-miR-375 promotes the progression of inflammatory bowel disease by upregulating TLR4.

OBJECTIVE: To elucidate the biological function of hsa-miR-375 in the progression of inflammatory bowel disease (IBD) and the potential mechanism. PATIENTS AND METHODS: Intestinal mucosa tissues of 26 IBD patients and 30 healthy volunteers who underwent colonoscopy were harvested for determining hsa-miR-375 level by quantitative Real-time polymerase chain reaction (qRT-PCR). Binding of hsa-miR-375 to toll-like receptor 4 (TLR4) was verified by the dual-luciferase reporter gene assay. Changes in the viability and apoptosis in Caco-2 cells influenced by hsa-miR-375 were examined by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. The regulatory effect of hsa-miR-375 on the intestinal epithelial barrier was examined by detecting transepithelial electrical resistance (TEER) and lucifer yellow flux. Relative levels of TLR4, nuclear factor-kappa B (NF-κB), zonula occludens-1 (ZO-1), occludin and inflammatory factors in Caco-2 cells were detected by qRT-PCR, Western blot and enzyme-linked immunosorbent assay (ELISA). RESULTS: Hsa-miR-375 was downregulated in intestinal mucosa tissues of patients with Crohn's disease (CD) and ulcerative colitis (UC). Knockdown of hsa-miR-375 decreased viability and TEER, but elevated apoptotic rate and lucifer yellow flux. Overexpression of hsa-miR-375 achieved the opposite trends. TLR4 was the direct downstream of hsa-miR-375, and its level was negatively mediated by hsa-miR-375. In addition, TLR4 level in Caco-2 cells was upregulated after LPS induction, while hsa-miR-375 level was unchangeable. Knockdown of hsa-miR-375 upregulated NF-κB and pro-inflammatory factors TNF-α, IL-1ß, IL-6 and IL-8, and downregulated ZO-1, occludin and anti-inflammatory factor IL-10. CONCLUSIONS: Hsa-miR-375 is involved in the pathogenesis of IBD by upregulating TLR4 and inducing NF-κB activation.

Long non-coding RNA GClnc1 promotes progression of colorectal cancer by inhibiting p53 signaling pathway.

OBJECTIVE: The aim of this study was to investigate whether long non-coding RNA (lncRNA) GClnc1 was involved in the development of colorectal cancer, and to explore its possible mechanisms. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was conducted to detect GClnc1 expression in 48 colorectal cancer tissues and normal colon tissues. The Kaplan-Meier method was used to analyze the relationship between GClnc1 expression and survival rate of patients with colorectal cancer. In addition, GClnc1 expression in colorectal cancer cell lines and normal colonic epithelial cell lines were analyzed. After knockdown and over-expression of GClnc1 in colorectal cancer cells, Cell Counting Kit-8 (CCK-8) and colony formation assay were performed to detect the viability and proliferation of cells, respectively. RNA pull-down and RNA-binding protein immunoprecipitation (RIP) were applied to examine the specific interaction between GClnc1 and p53. After over-expression of GClnc1 in colorectal cancer cells, qPCR and Western blot were performed to evaluate the expression levels of p53, p21 and BAX. Meanwhile, the Luciferase reporter gene assay was established to reveal the activity of p53 after over-expression of GClnc1. ChIP assay was applied to figure out whether GClnc1 could affect the binding ability of p53 to the promoter region of p21. After p53 or GClnc1 knock-down in colorectal cancer cells, the protein level of p53 was analyzed using Western blot. Finally, qRT-PCR, CCK-8 and colony formation assay were used to detect the levels of p21 and BAX, the viability, as well as the proliferation ability of cells, respectively. RESULTS: The expression of GClnc1 in colorectal cancer tissues was significantly higher than that of para-cancerous tissues. Meanwhile, GClnc1 expression in T3 and T4 tumors was markedly higher than that of T1 and T2. The survival analysis revealed that patients with a higher level of GClnc1 showed remarkably lower overall survival than those with lower expression of GClnc1. QRT-PCR results indicated that GClnc1 expression in colorectal cancer cells (including SW620 and HCT116) was conspicuously higher than that of normal colonic epithelial cells (NCM640). After knocking down GClnc1 in SW620 cells, the viability and proliferation abilities were conspicuously decreased. Meanwhile, the expression level of GClnc1, as well as the viability and colony formation ability of cells, were significantly increased after over-expression of GClnc1 in HCT116 cells. Subsequently, the qRT-PCR assay demonstrated that GClnc1 was mainly localized in the nucleus. RNA pull-down and RIP experiments revealed that there was a specific interaction between GClnc1 and p53. Moreover, qRT-PCR and Western blot analysis indicated that the expression level of p53 was not affected after over-expression of GClnc. However, the expressions of p21 and BAX were remarkably decreased. The Luciferase reporter gene assay revealed that GClnc1 over-expression markedly weakened the Luciferase activity of p53. Meanwhile, ChIP experiments demonstrated that GClnc1 up-regulation affected the binding condition of p53 to p21. Western blot analysis showed that knockdown of p53 reversed the increased mRNA level of p21 as well as BAX. Furthermore, p53 down-regulation significantly weakened cell viability and colony formation ability caused by knockdown of GClnc1. CONCLUSIONS: LncRNA GClnc1 was highly expressed in colorectal cancer tissues. Meanwhile, it could increase the proliferation of colorectal cancer cells by reducing the expression of p21 as well as BAX via p53 signaling pathway, thereby promoting the progression of colorectal cancer.

[Clinical effects of application of antibiotic bone cement in wounds of diabetic foot ulcers].

Objective: To explore the clinical effects of antibiotic bone cement in the treatment of diabetic foot ulcers. Methods: According to the treatment methods, 18 patients with diabetic foot ulcers (11 males and 7 females, aged 53-79 years), who were conformed to the study criteria and admitted to our hospital from January 2016 to January 2017, were enrolled in traditional group; 18 patients with diabetic foot ulcers (11 males and 7 females, aged 55-80 years), who were conformed to the study criteria and admitted to our hospital from February 2017 to February 2018, were enrolled in bone cement group. Wounds of patients in traditional group were treated with vacuum sealing drainage after conventional debridement. Wounds of patients in bone cement group were covered with antibiotic bone cement after conventional debridement. The number of patients with positive bacterial culture in wound exudate in the 2 groups on admission and 3, 6, 9, and 15 days after surgery, the length of hospital stay, the number of operation, and the wound complete healing time were retrospectively recorded. Data were processed with Fisher's exact probability test and independent sample t test. Results: Compared with (29±10) d and (4.6±1.2) times of patients in traditional group, the length of hospital stay [(9±3) d] of patients was obviously shortened, the number of operation [(1.3±0.6) times] of patients was obviously reduced, the number of patients with positive bacterial culture in wound exudate at each time point post surgery was obviously reduced (t=8.177, 9.896, P<0.05 or P<0.01) in bone cement group. There were no statistically significant differences in the number of patients with positive bacterial culture in wound exudate on admission and wound complete healing time between patients in the 2 groups (t=0.175, P>0.05). Conclusions: The antibiotic bone cement treatment of diabetic foot ulcers can reduce the number of patients with positive bacterial culture in wound exudate and the number of operation, as well as shorten the length of hospital stay.