Objective : To investigate the effect of laminarin(LAM) on nonproliferative diabetes retinopathy by high throughput sequencing(RNA-seq). Methods : The diabetes model was established by intraperitoneal injection of streptozotocin(STZ), and the effect of LAM on diabetic mice was observed.C57BL/6 mice were randomly divided into three groups: Control group, Model group, and LAM group, with 8 mice in each group. After 8 weeks of modeling, the LAM group received a 4-week intraperitoneal injection of LAM treatment. Changes in blood glucose and body weight of the three groups of mice were recorded, HE staining was performed to examine retinal lesions, and RNA-seq was used to identify differentially expressed genes(DEGs) in diabetic retinopathy(DR) under the action of STZ and LAM. Results : STZ successfully established the model of DR, and LAM reduced the blood sugar in diabetic mice to a certain extent and improved the pathological morphology of retinal structural looseness in diabetic mice. After RNA-seq analysis of DEGs, it was found that there were a total of 214 DEGs in the retina of the Model group mice compared to the Control group. Enrichment analysis revealed that DR could exacerbate the lesions through the PI3K Akt signaling pathway. There were a total of 42 DEGs in the retina of the Model group and LAM group mice, and enrichment showed that LAM improved the lesions through the neutrophil extracellular trap pathway. Early growth response factor 1(Egr1), FBJ osteosarcoma oncogene(Fos), nuclear receptor subfamily 4A member 1(Nr4a1), and salt-induced kinase 1(Sik1) were regulated by STZ, and LAM significantly regulated their expression, which might be closely related to LAM′s treatment of diabetic retinopathy. Conclusion DEGs can exacerbate the severity of diabetic retinopathyviathe PI3K-Akt signaling pathway. LAM can mitigate diabetic retinopathyviathe neutrophil extracellular trap pathway. Egr1, Fos, Nr4a1, and Sik1 are key genes involved in LAM treatment of STZ-induced DR.

Objective To explore the correlation of serum Chemerin level with disease activity and the ratio of T helper 17/regulatory T cells(Th17/Treg)in patients with rheumatoid arthritis(RA).Methods A total of 180 patients with RA who were admitted to our hospital were regarded as the observation group.According to the DAS28 score,the observation group was divided into the high activity group(60 cases),the moderate activity group(60 cases)and the low activity group(60 cases).Another 180 healthy people who underwent physical examination in our hospital during the same period were regarded as the control group.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum levels of Chemerin,interleukin-9(IL-9),interleukin-10(IL-10)and interleukin-17(IL-17).Flow cytometry was used to detect the Th17/Treg ratio.Spearman correlation analysis was applied to analyze the correlation between serum Chemerin level and DAS28 score.Pearson correlation analysis was used to analyze the correlation between serum Chemerin level and Th17,Treg cell percentage and Th17/Treg ratio.Results The results of this study showed that the serum level of Chemerin was higher in the observation group than that in the control group(P＜0.05).The serum Chemerin level was positively correlated with DAS28 score(P＜0.05).Serum Chemerin levels and DAS28 scores decreased in turn in the high,moderate and low activity groups(P＜0.05).The percentage of Th17 cells and the ratio of Th17/Treg were higher in the observation group than those in the control group,and the percentage of Treg cells was lower in the observation group than that in the control group(P＜0.05).The level of IL-10 was lower in the observation group than that in the control group,while levels of IL-17 and IL-9 were higher in the observation group than those in the control group(P＜0.05).The results of Pearson correlation analysis showed that serum Chemerin level was positively correlated with the percentage of Th17 cells and the ratio of Th17/Treg,and negatively correlated with the percentage of Treg cells(P＜0.05).Conclusion Serum Chemerin level is elevated in patients with RA,which is closely related to disease activity and Th17/Treg ratio.

Objective:To explore the rapid evaluation of the early pathogen of severe Chlamydophila psittaci pneumonia by bedside diagnostic bronchoscopy, so as to start effective anti-infection treatment before the results of macrogenome next generation sequencing (mNGS) test. Methods:The clinical data of three patients with severe Chlamydophila psittaci pneumonia who were successfully treated in the First Affiliated Hospital of Xinjiang Medical University, the First People's Hospital of Aksu District, and the First Division Hospital of Xinjiang Production and Construction Corps from October 2020 to June 2021 were retrospectively analyzed, including the rapid assessment of early pathogens by bedside diagnostic bronchoscopy and the use of antibiotics to start anti-infection treatment. These patients were successfully treated. Results:The three patients were male, aged 63, 45 and 58 years old, respectively. Before the onset of the penumonia, they had a clear medical history of bird exposure. The clinical manifestations mainly included fever, dry cough, shortness of breath and dyspnea. One case had abdominal pain and lethargy. The results of laboratory examination indicated that the peripheral blood white blood cell count (WBC) of two patients were high [(10.2-11.9)×10 9/L], the percentage of neutrophils increased (85.2%-94.6%) and the percentage of lymphocytes decreased (3.2%-7.7%) in all 3 patients after admission to hospital and entering into intensive care unit (ICU). The procalcitonin (PCT) of 3 patients increased after admission, and still increased when entering ICU (0.3-4.8 ng/L), so did C-reactive protein (CRP, 58.0-162.0 mg/L) and erythrocyte sedimentation rate (ESR, 36.0-90.0 mm/1 h). After admission, serum alanine transaminase (ALT) increased in 2 cases (136.7 U/L, 220.5 U/L), so did aspartate transaminase (AST) in 2 cases (249.6 U/L, 164.2 U/L). ALT (162.2-267.9 U/L) and AST (189.8-223.2 U/L) increased in 3 patients when they entered ICU. The level of serum creatinine (SCr) of 3 patients were normal after admission and entering ICU. The chest computed tomography (CT) findings of 3 patients were acute interstitial pneumonia, bronchopneumonia and lung consolidation, of which 2 cases were accompanied by a small amount of pleural effusion, and 1 case was accompanied by more regular small air sacs. Multiple lung lobes were involved, but mainly one lung lobe. The oxygenation index (PaO 2/FiO 2) of the 3 patients admitting to ICU were 100.0, 57.5 and 105.4 mmHg (1 mmHg ≈ 0.133 kPa), respectively, which met with the diagnostic criteria of moderate and severe acute respiratory distress syndrome (ARDS). All three patients received endotracheal intubation and mechanical ventilation. Under the bedside bronchoscope, the bronchial mucosa of 3 patients were obviously congested and edematous, without purulent secretion, and there was 1 case with mucosal hemorrhage. Three patients underwent bedside diagnostic bronchoscopy, and the evaluation result of the pathogen was that it might be atypical pathogen infection, so they were given moxifloxacin, cisromet and doxycycline intravenously, respectively, and combined with carbapenem antibiotics intravenously. After 3 days, the detection results of mNGS in bronchoalveolar lavage fluid (BALF) showed that only Chlamydia psittaci was infected. At this time, the condition was significantly improved, and PaO 2/FiO 2 was significantly increased. Therefore, the antibiotic treatment scheme remained unchanged, and mNGS only served to verify the initial diagnosis. Two patients were extubated on the 7th and 12th day of admission to the ICU, respectively, while one patient was extubated on the 16th day of admission to the ICU due to nosocomial infection. All 3 patients were transferred to the respiratory ward after the condition was stable. Conclusion:The bedside diagnostic bronchoscopy based on clinical characteristics is conducive to not only the rapid assessment of the early pathogens of severe Chlamydia psittaci pneumonia, but also effective anti-infection treatment before the returning of mNGS test results, which can make up for the lag and uncertainty of the mNGS test results.

Objective: To investigate the effects of interleukin ( IL) -10 on the proliferation of HaCaT cells and CaCl2 induced expression of differentiation markers and its possible molecular mechanisms． Methods: HaCaT cells were treated with various concentrations of IL-10 (0，3，10，30 ng / ml) for different time (0，24，48，72 h) ，cell proliferation was measured using MTS，and cell cycle was determined by flow cytometry．HaCaT cells were pretreated with IL-10 (final concentration 10 ng / ml) for 1 h，then incubated with or without CaCl2 (final concentration 1. 2 mmol / L) for 24，48，72 h ，Western blot was performed to detect the effect of IL-10 on the expression of HaCaT keratinocyte differentiation markers．After pretreatment of HaCaT cells with PD98059，an inhibitor of mitogen-activated kinase-ERK1 /2，and LY294002，an inhibitor of phosphatidylinositol kinase-serine / threonine kinase (PI3K-AKT) ，the total RNA and proteins were extracted separately，real time quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to examine the influence of IL-10 on the expression of differentiation markers (Keratin1，Keratin5，Involucrin) . Results : MTS results revealed that IL-10 (30 ng / ml and lower doses) did not alter the proliferation of HaCaT cells in 72 h．Flow cytometry analysis demonstrated that IL-10 had no significant influence on cell cycle progression．The results of Western blot showed that IL-10 upregulated the expression of differentiation markers Involucrin，while there was no significant effect on Keratin1 and Keratin5 ．Mechanism research analysis demonstrated that IL-10 could activate ERK1 /2 and AKT ，increase their phosphorylation levels ; RT-qPCR and Western blot results showed that PD98059 and LY294002 partially blocked IL-10 induced Involucrin expression． Conclusion At a particular concentration range，IL-10 has little effect on HaCaT proliferation ，but it partially upregulates the expression of differentiation marker Involucrin via the MAPKs-ERK1 /2 and PI3K-AKT pathways.

Objective:To observe the effect of Ba Duan Jin(Eight-brocade Exercise)plus Tuina(Chinese therapeutic massage)in treating primary dysmenorrhea due to cold-induced blood stasis in female college students and on the score of fatigue scale-14(FS-14). Methods:Seventy-two female college students with primary dysmenorrhea due to cold-induced blood stasis were randomized into a Tuina group and a joint group,with 36 cases in each group.The Tuina group only received Tuina manipulations.In the joint group,besides the same Tuina manipulations,patients practiced Ba Duan Jin.For both groups,the once-daily intervention was conducted from 6 d before the menstrual period until menstrual day 1 for 3 menstrual cycles.Changes in the scores of COX menstrual symptom scale(CMSS),visual analog scale(VAS),and FS-14 after the intervention were observed.Clinical efficacy was also estimated. Results:During the process,1 case dropped out in the Tuina group,and 35 cases completed the intervention;2 cases dropped out in the joint group,and 34 cases completed the intervention.The total effective rate was 94.1%in the joint group,higher than 88.6%in the Tuina group(P<0.05).After treatment,the symptom duration and intensity scores in the scores of CMSS,VAS,and FS-14 declined in both groups(P<0.05 or P<0.01);the CMSS symptom duration score and FS-14 score were lower in the joint group than in the Tuina group(P<0.05). Conclusion:Tuina manipulations alone or combined with Ba Duan Jin practice can effectively treat primary dysmenorrhea due to cold-induced blood stasis in female college students;when combined with Ba Duan Jin practice,Tuina manipulations can more significantly improve pain duration and fatigue,suggesting the advantages of combining Tuina Gongfa and manipulations.

Objective: To screen the differential genes and pathways of female and male C57 BL/6 J mice by transcriptome sequencing, and to lay a foundation for further exploring the molecular mechanism of behavioral differences between male and female mice. Methods: 8-week-old female and male mice of the C57 BL/6 J strain were completely isolated from the mouse hippocampus, and total RNA was extracted and reverse transcribed to construct a cDNA library, and 50 single-ended mice were generated on the BGIseq500 platform(BGl-Shenzhen, China) The base reads were transcriptome sequenced. After sequencing, based on GO and KEGG databases, combined with the phyper function in R language to screen, correct and enrich the data, calculatePvalue, and then perform FDR correction onPvalue to obtainQvalue and analyze different annotated genes based on the hypergeometric test method. The expression status was analyzed, and the differential genes and pathways in the hippocampus of mice of different genders were screened out. Results: Through the comparison of male and female differences, 325 differential genes were screened, including 233 up-regulated genes and 92 down-regulated genes. The functions of these differential genes were mainly enriched in long-term potentiation(LTP), calcium signal pathway, nicotine addiction and other processes. There were 362 junctions and 1 703 interaction edges in the female-male differential gene interaction network, and 10 core genes selected: lysine demethylase 5 D(Kdm5 d), cyclin-dependent kinase 5(Cdkl5), Cell output mediator(Cle1), dopamine receptor 6 a3(Slc6 a3), cassette transcription factor(Fox), precursor mRNA processing factor 4 B(Prpf4 b), glycine receptor 4(Glur4), calcium ion receptor 2(Camk2), DNA and RNA binding protein family(Son), serotonin receptor 5 b(Htr5 b). Conclusion The selected differential genes and signal pathways may lay the foundation for explaining the molecular mechanism of the differences in behavior between male and female mice.

Objective : To investigate the effects of RO or incubation with cholesterol ( CH) on the differentiation and apoptosis of KCs. Methods : Ca2 + ( 1. 8 mmol/L) was added to KCs for 1 day after co - culture of RO alone or combined with CH in KCs for different time. The expression of Involucrin (INV) and Loricrin in KCs was analyzed by Western blot. After co⁃culture of RO alone or combination with CH in KCs for different time , the apoptotic changes of KCs cells were detected by flow cytometry and the expression of apoptotic proteins Bax and Bcl⁃2 were verified by Western blot. Results : RO down⁃regulated the expression of Ca2 + induced differentiation marker protein INV , but weakly inhibited the expression of Loricrin , while CH showed no antagonistic effect on RO. RO induced apoptosis of KCs cells in a time dependent manner. CH could antagonize the apoptotic effect of RO on KCs ; the expression of Bcl⁃2 and Bax was inhibited when RO was applied alone , CH could partially antagonize the inhibition effect of RO on Bcl⁃2 expression , but had no significant effect on Bax; however, RO reduced the ratio of Bcl⁃2/Bax in a time⁃dependent manner, and CH partially weakened the effect of RO on the ratio of Bcl⁃2/Bax. Conclusion RO may inhibit KCs differentiation and induce KCs cell apoptosis by down⁃regulating the expression of Loricrin and Bcl⁃2.

Objective : To investigate the molecular changes of lanosterol synthase(LSS) gene dysfunction and cholesterol levels that may affect brain function. Methods: The differentially expressed genes in LSS knockout(+/-) mice and WT C57 BL/6 mice were analyzed by RNA-SEQ,and the signal pathways involved in the differentially expressed genes were analyzed by KEGG enrichment. The differential genes were detected by real-time quantitative PCR and the protein expression of differential genes was detected by Western blot. Results: By RNA-SEQ analysis, LSS(+/-) heterozygous knockout resulted in the difference of 320 genes in the mouse brain, including 145 up-regulated genes and 175 down-regulated genes.KEGG enrichment analysis showed that these differentially expressed genes involved in neural-activated receptor and ligand pathways, hippocampal signaling pathways and glutamate synaptic pathways.Real-time quantitative PCR showed that CCKBR and Htr5 b were up-regulated, respectively. Western blot results showed that CCKBR gene was highly expressed in the total protein of all brain tissues of LSS knockout(+/-) mice. Conclusion LSS heterozygotic knockout may affect brain function and animal behavior.

Objective:To explore the clinical application of free chimeric medial femoral condyle osteofascial free flap (CMFCOF) in the treatment of traumatic composite bone and soft tissue defect of hand and foot.Methods:Between January, 2015 and March, 2020, 8 patients with traumatic composite bone and soft tissue defect in hand and foot were treated with CMFCOF. Of the 8 patients, there were 6 males and 2 females, with an average age of 41 (range, 24 to 56) years. The causes of injury included 3 of traffic accident, 3 of machine crush and 2 of crush. Two cases had proximal phalanx defect, 3 with metacarpal bone and 3 with metatarsal bone. The time between injury to the flap repair were 2 to 120 (mean, 84) days. The size of bone defect ranged from 2.0 cm×1.2 cm×1.2 cm to 4.4 cm× 3.0 cm×2.3 cm. The soft tissue defect ranged from 2.0 cm×1.4 cm to 5.6 cm×4.5 cm. All bone defects were on the diaphysis, without involvement of joints. Two cases had tendon defect. According to the defect of bone and soft tissue, the CMFCOF was prepared and skin graft was performed on the surface of its fascial flap.Results:The average time of flap harvesting was 53(52-96) minutes. All donor sites were directly closed. All flaps and skin grafts achieved stage I survival. All patients entered 9-16 months of follow-up, with an average of 14.5 months. The average healing time of bone was 7.5 (range, 6-10) weeks. At the last follow-up review, all flaps were not thinned. The function of donor site was restored well, without weight bearing disorder and paraesthesia in the anterior patella area. According to the trial standard of Digit Function Evaluation of the Hand Surgery Society of Chinese Medical Association, 3 patients were rated as excellent, 1 was good and 1 was fair. According to the Maryland foot evaluation criteria, 3 patients were rated as excellent for recovered with normal weight-bearing walking.Conclusion:CMFCOF can achieve satisfactory results in repairing composite bone and soft tissue defect of hand or foot. The flap has the advantages in simple operation, high quality of bone and concealed donor site.