Objective : To construct a human keratinocyte-forming cell line(HaCaT) with stable knockout of the G protein-coupled receptor 107(GPR107) gene, and to preliminarily investigate the effect of GPR107 deletion on the biological behaviour of HaCaT cells. Methods : Using CRISPR/Cas9 gene editing technology, HaCaT cells with knockout ofGPR107gene were constructed and monoclonal cells with GPR107 deletion were obtained by limited dilution method. Genomic DNA was amplified using Western blot and PCR and sequenced to validate the single-cell clones with knockdown of GPR107. The cell cycle changes were detected by flow cytometry; cell proliferation was detected by CCK-8; apoptosis was detected by flow cytometry; changes in cell differentiation markers were detected by Western blot; cell migration ability was analyzed by cell scratch assay and other methods. Results : LentiCas9-Blast and plenti-guide-RNA-GPR107 plasmids were successfully transfected into HaCaT cells, 21 monoclonal cell lines were obtained by limited dilution, and Western blot showed that the GPR107 expression was significantly reduced in 8 of them; PCR sequencing of the cellular genome was used, which resulted in the obtainment of C4 and 2D8GPR107-/-HaCaT monoclonal cell lines. CCK-8 assay and flow cytometry assay showed thatGPR107gene deletion resulted in G0G1phase block, significantly weakened proliferation ability and increased apoptosis level of HaCaT cells. Western blot found that the differentiation of HaCaT cells accelerated after knockdown ofGPR107. Additionally the results of the cell scratch assay indicated that the migration ability of HaCaT cells was enhanced after knockdown ofGPR107. The results showed that the migration ability of HaCaT cells was enhanced after knockdown ofGPR107. Conclusion HaCaT cell line withGPR107gene deletion is successfully constructed, GPR107 deletion blocks the G0G1phase of HaCaT cells, which inhibiting the proliferation of HaCaT cells and promoted apoptosis, and it was found that the differentiation and migration of HaCaT cells were enhanced after knocking downGPR107.

Objective To investigate the role and mechanism of urate in chronic kidney disease complicated with renal interstitial fibrosis(CKD-RIF).Methods Mice were continuously fed with a diet containing 0.2％adenine for a duration of 9 weeks to establish mice models with CKD-RIF.By the end of the 9-week experimental periods,collected blood samples from the posterior orbital venous plexus of mice to measure renal functions and serum urate concentrations prior to euthanizing the mice.Hematoxylin-eosin(HE)staining and periodic acid-Schiff staining(PAS)were used to investigate the pathological alternations in kidney tissues.Masson's trichrome staining was used to observe the extent of renal fibrosis.Urate staining was used to detect urate deposition in renal tissues.Western blot and immunohistochemistry were used to detect the expression of target molecules.Scratch tests were used to ex-amine the migration abilities of cells treated with different concentrations of uric acid.Results The kidney function analysis showed that a significant increase in the levels of serum urea nitrogen(P=0.006 4),creatinine(P=0.008 0)and urate(P=0.000 7)in the CKD-RIF mice compared with the normal control group.The results of HE staining and PAS staining showed a significance of renal tubule injury and infiltration of inflammatory cells in the model group.Masson's trichrome staining showed that a marked increase in collagen deposition in the model group.The results of urate staining showed a significant presence of urate crystals in kidney tissue of the model group when compared to the control group.Animal tissue immunoblotting and immunohistochemistry analysis showed a significant increase in the expression levels of vimentin,α-SMA and TGF-β1 in the model group in comparison to the control group.Conversely,in the model group,E-cadherin levels exhibited a dramatic reduction compared to the control group.The findings from the scratching tests showed that uric acid significantly enhanced cell migration.Western blot analysis showed a dramatic increase in the expression levels of vimentin and α-SMA,while E-cadherin exhibited significant decrease in the cells subjected to uric acid treatment.Conclusion Urate stimulates the secre-tion of TGF-β1 by renal tubule epithelial cells and induces epithelial-mesenchymal transdifferentiation,thereby ex-acerbating renal interstitial fibrosis in CKD.

Objective: To investigate the role and mechanism of Sigirr deletion in chronic kidney disease complicated with renal interstitial fibrosis (CKD⁃RIF) in mice. Methods: polymerase chain reaction (PCR) was used for identification of gene types of mice. Mice were continuously fed with the foods containing 0. 2% adenine for 12 weeks to establish the CKD⁃RIF models. Then , serum was collected to detect levels of creatinine and nitrogen when mice were killed. H&E staining was used to analyze the pathological changes of kidney tissues. Masson staining was used to observe the degree of renal fibrosis. Immunohistochemistry was used to detect the changes of the interest proteins , such as IL⁃1β , MyD88 , activated NF⁃κB , TGF⁃ β1 , E ⁃cadherin and Vimentin. Results: Serum creatinines and urea nitrogens of mice fed with high adenine (CKD⁃RIF groups) significantly increased , compared with those of the control groups. H&E and Masson staining results showed that there were more infiltrated inflammatory cells and more critical collagen fiber deposition in the renal tissues of the Sigirr - / - mice with CKD⁃RIF. Western blot and Immunohistochemical analysis showed that the expression of IL⁃1β and its downstream MyD88 increased , and the level of phosphorylated NF⁃κB (p⁃P65) significantly increased in the renal tissues of CKD⁃RIF mice compared with the controls. And upregulation of these proteins in renal tissues of Sigirr - / - mice with CKD⁃RIF was more obvious than that of the CKD⁃RIF Sigirr + / + mice. TGF⁃ β1 , as a key cytokine involved in renal interstitial fibrosis , significantly increased ,followed by the increase of vimentin , as well as the decrease of E ⁃cadherin . The results of vimentin and cadherin E detected by Western blot were consistent with those of immunohistochemistry , and α ⁃SMA also increased significantly. Conclusion Adenine diet successfully induces CKD⁃RIF mice models. Sigirr deletion is beneficial to activation of the IL⁃1β mediating NF⁃κB signal pathway ,which promotes TGF⁃ β1 expression in the renal interstitiums to induce renal interstitial fibrosis.

The aquatic grass Zizania latifolia grows symbiotically with the fungus Ustilago esculenta producing swollen structures called Jiaobai, widely cultivated in China. A new disease of Z. latifolia was found in Zhejiang Province, China. Initial lesions appeared on the leaf sheaths or sometimes on the leaves near the leaf sheaths. The lesions extended along the axis of the leaf shoots and formed long brown to dark brown streaks from the leaf sheath to the leaf, causing sheath rot and death of entire leaves on young plants. The pathogen was isolated and identified as the bacterium Pantoea ananatis, based on 16S ribosomal RNA (rRNA) gene sequencing, multilocus sequence analysis (atpD (β-subunit of ATP synthase F1), gyrB (DNA gyrase subunit B), infB (translation initiation factor 2), and rpoB (β‍-subunit of RNA polymerase) genes), and pathogenicity tests. Ultrastructural observations using scanning electron microscopy revealed that the bacterial cells colonized the vascular tissues in leaf sheaths, forming biofilms on the inner surface of vessel walls, and extended between vessel elements via the perforated plates. To achieve efficient detection and diagnosis of P. ananatis, species-specific primer pairs were designed and validated by testing closely related and unrelated species and diseased tissues of Z. latifolia. This is the first report of bacterial sheath rot disease of Z. latifolia caused by P. ananatis in China.
Pantoea/genetics* ; Plant Diseases/microbiology* ; Poaceae/microbiology* ; Virulence

Objective    To compare the clinical effects of segmentectomy and lobectomy for ≤2 cm lung adenocarcinoma with micropapillary and solid subtype negative by intraoperative frozen sections. Methods    The patients with adenocarcinoma who received segmentectomy or lobectomy in multicenter from June 2020 to March 2021 were included. They were divided into two groups according to a random number table, including a segmentectomy group (n=119, 44 males and 75 females with an average age of 56.6±8.9 years) and a lobectomy group (n=115, 43 males and 72 females with an average of 56.2±9.5 years). The clinical data of the patients were analyzed. Results    There was no significant difference in the baseline data between the two groups (P>0.05). No perioperative death was found. There was no statistical difference in the operation time (111.2±30.0 min vs. 107.3±34.3 min), blood loss (54.2±83.5 mL vs. 40.0±16.4 mL), drainage duration (2.8±0.6 d vs. 2.6±0.6 d), hospital stay time (3.9±2.3 d vs. 3.7±1.1 d) or pathology staging (P>0.05) between the two groups. The postoperative pulmonary function analysis revealed that the mean decreased values of forced vital capacity and forced expiratory volume in one second percent predicted in the segmentectomy group were significantly better than those in the lobectomy group (0.2±0.3 L vs. 0.4±0.3 L, P=0.005; 0.3%±8.1% vs. 2.9%±7.4%, P=0.041). Conclusion    Segmentectomy is effective in protecting lungs function, which is expected to improve life quality of patients.

Objective:To assess the correlation between frailty and cardiac autonomic nervous system function in elderly patients.Methods:Elderly hospitalized patients aged 65 years and over were enrolled and assessed for frailty by using the clinical frailty scale.Cardiac autonomic modulation was evaluated by heart rate variability analysis through 24 h electrocardiogram recording.Results:A total of 180 elderly patients were enrolled in this study, including 66 patients with frailty and 114 patients without frailty.The mean age of the frailty group was higher than that of the non-frailty group(79.8±6.0 vs.75.0±6.3, t=5.030, P<0.001). The proportions of patients with hypertension, stroke/transient cerebral ischemia attack(TIA), heart failure and osteoarthritis were higher in the frailty group than in the non-frailty group(all P<0.05). Compared with the non-frailty group, the standard deviation of normal-to-normal intervals(SDNN)[103.0(76.0, 121.2) vs.107.5(92.0, 136.0), Z=-2.108, P=0.035], the standard deviation of the averages of NN intervals in all 5-min segments(SDANN)[86.0(67.7, 106.5) vs.97.5(78.0, 126.0), Z=-2.694, P=0.007], normalized low frequency(LFnorm)(53.1±13.0 vs.59.3±13.9, t=-3.024, P=0.003)and low frequency/high frequency(LF/HF)ratio[1.2(1.0, 1.4) vs.1.4(1.1, 1.7), Z=-3.041, P=0.002]were decreased and normalized high frequency(HFnorm)(36.8±9.2 vs.32.2±10.7, t=3.033, P=0.003)was increased in the frailty group.HFnorm in the frailty group was significantly higher than that in the non-frailty group.The incidents of SDANN<92 ms, LFnorm<50 nU, HFnorm>32 nU and LF/HF ratio<1.5 were higher in the frailty group than in the non-frailty group(59.1% or 39/66 vs.41.2% or 47/114, 42.4% or 28/66 vs.22.8% or 26/114, 72.7% or 48/66 vs.49.1% or 56/114, 84.8% or 56/66 vs.65.8% or 75/114, χ2=5.346, 7.660, 9.547, 7.664, P=0.021, 0.006, 0.002, 0.006). Logistic multivariate regression analysis showed that LFnorm, HFnorm and LF/HF ratio were correlated with frailty( OR=0.971, 1.039 and 0.333, all P<0.05), and HFnorm>32 nU and LF/HF ratio<1.5 were risk factors for frailty( OR=2.401 and 2.773, both P<0.05). Conclusions:Cardiac autonomic nerve system function is impaired in elderly frail patients, with the imbalance between the sympathetic and vagus nerves.Therefore particular attention should be paid to heart rate variability in elderly patients with frailty.

Objective:To evaluate the biomechanical performance of our self-designed anatomical plate for posterolateral tibial plateau in comparison with conventional plates for treatment of posterolateral tibial plateau fractures.Methods:A novel anatomic plate for posterolateral tibial plateau was designed according to the data measured in the superior fibular capitulum and 3D CT segmentation. Twenty-four knee joints were obtained from 12 freshly frozen adult cadavers to make models of posterolateral tibial plateau fracture. The models were divided into 3 groups( n=8). In group A, fixation was simulated via the supra-fibular-head approach after autogenous iliac bone-graft by our self-designed anatomic plate for posterolateral tibial plateau; in group B, fixation was simulated via the posterior tibial approach after autogenous iliac bone-graft by a small T-plate; in group C, fixation was simulated via the supra-fibular-head approach after autogenous iliac bone-graft by a normal L-plate. Biomechanical tests were carried out in the 3 groups to measure the vertical displacements of split bone fragment under the vertical compression loads of 500 N, 1,000 N and 1,500 N and the maximum compression upon failure of internal fixation (compressed displacemen t=3 mm). Results:At the vertical compression loads of 500 N, 1,000 N and 1,500 N, the vertical displacements of split bone fragment showed significant differences among the 3 groups ( P<0.05); there was a significant difference between group C and groups A and B, respectively ( P<0.05), but an insignificant difference between group A and group B ( P>0.05) though group A performed slightly better. In terms of the maximum compression upon failure of internal fixation, significant differences existed among the 3 groups ( P<0.05); there was a significant difference between group C and groups A and B, respectively ( P< 0.05), but an insignificant difference between group A and group B ( P>0.05). Conclusions:Our self-designed anatomic plate for posterolateral tibial plateau can firmly fixate the fracture fragments of posterolateral condyle.

Objective:To assess the cardiac autonomic nervous function in elderly patients with frailty.Methods:Patients aged ≥ 65 years old admitted in Beijing Hospital from September 2018 to August 2019 were enrolled in this study. Clinical frailty score was used to assess the frailty. The cardiac autonomic modulation was evaluated by sinus heart rate turbulence analysis through 24 h electrocardiogram recording.Results:A total of 129 elderly patients were finally enrolled in this study with a mean age of (77.5±6.4) years, 58.1% of them were male. There were 53 patients in frail group and 76 patients in non-frail group. The age of the frailty group was significantly higher than that of the non-frailty group [(80.5±5.5) vs.(75.3±6.2)]; the prevalence of hypertension [84.9%(45/53)], heart failure [32.1%(17/53)] and peripheral vascular diseases [32.1%(17/53)] in the frailty group was significantly higher than that in the non-frailty group [65.8%(50/76), 1.3%(1/76), 17.1%(13/76); t=5.001, χ 2=5.879, 24.606, 3.921; all P<0.05]. Compared with non-frailty group, turbulence onset (TO) [-0.05(-0.92, 0.82)% vs. -0.74(-1.58, 0)%; Z=2.616, P=0.009] was significantly higher in frailty group, while turbulence slope (TS) [2.34(1.30, 5.00)ms/RR vs. 4.34(2.66, 6.39)ms/RR; Z=-3.048, P=0.002] was significantly lower. The rate of TO abnormality [49.1% (26/53) vs. 26.3%(20/76), χ 2=7.038, P=0.008] and TS abnormality [34.7%(29/53) vs. 21.0%(16/76); χ 2=15.579, P<0.001] in the frailty group was significantly higher than that in the non-frailty group. Multivariate logistic regression analysis showed that TO abnormality( OR=2.970, P=0.010, 95 %CI:1.300-6.785) and TS abnormality( OR=3.618, P=0.003, 95 %CI:1.565-8.364) were correlated with frailty. Conclusion:Cardiac autonomic nerve function may be impaired in elderly frail patients, and decreased vagal nerve tension may be presented.

Objective :To explore therapeutic effect of upper and lower extremity active and passive rehabilitation treadmill combined occupational therapy on patients with chronic obstructive pulmonary disease (COPD) during sta—ble period .Methods : A total of 92 COPD patients were randomly and equally divided into occupational therapy group and treadmill + occupational therapy group .Both groups received routine nursing care ,treatment and com—prehensive rehabilitation therapy ,occupational therapy group also received occupational therapy ,while treadmill +occupational therapy group received upper and lower extremity active and passive treadmill training based on occupa—tional therapy group .After eight—week treatment ,pulmonary function ,exercise function ,daily living capacity and quality of life were analyzed and compared between two groups .Results : Compared with before treatment ,after treatment ,there were significant improvements in pulmonary function ,exercise function ,score of activity of daily living scale (ADL) and quality of life (SGRQ) in two groups , P＜ 0. 05 or ＜ 0. 01. Compared with occupational therapy group after treatment ,there was significant reduction in modified Medical Research Council dyspnea scale (mMRC) [ (2. 7 ± 0. 4) grade vs.(2. 4 ± 0.6) grade] ,and significant rise in 6min walking distance [ (291. 4 ± 28. 9) m vs.(307. 8 ± 30. 4) m] and ADL score [(56.0 ± 11.4) scores vs .(62. 0 ± 10.9) scores] in treadmill + occupation—al therapy group ( P＜0.05 or ＜0. 01) ,but there were no significant differences in pulmonary function indexes and quality of life (SGRQ) between two groups , P＞0.05 all.Conclusion : Upper and lower extremity active and passive treadmill training combined occupational therapy can significantly improve pulmonary function and exercise function and daily living capacity in patients with chronic obstructive pulmonary disease .