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 correlation between neutrophil lymphocyte ratio(NLR),IL-6 and lymphocyte subsets and prognosis of viral pneumonia complicated with acute respiratory distress syndrome(ARDS).Methods:A total of 98 patients with viral pneumonia complicated with ARDS admitted to Xinxiang Central Hospital/the Fourth Clinical College from May 2017 to June 2019 were selected,NLR,IL-6 and lymphocyte subsets expressions in patients with different severity were analyzed.Correlation between NLR,IL-6 and lymphocyte subsets levels in prognosis of viral pneumonia complicated with ARDS was analyzed by Pearson correlation.Logistc regression was used to analyze prognostic risk factors of viral pneumonia complicated with ARDS.Results:With aggravation of viral pneumonia complicated with ARDS,NLR and IL-6 levels were significantly increased(P<0.05),while CD3+T,CD4+T and CD19+T cells levels were significantly decreased(P<0.05).Pearson correlation analysis results showed that NLR and IL-6 were positively correlated with prognosis of viral pneumonia complicated with ARDS(P<0.05),while CD3+T,CD4+T,CD19+T and CD8+T cells were negatively correlated with prognosis of ARDS with viral pneumonia(P<0.05).Multivariate Logistic analysis showed that NLR,IL-6,CD3+T and CD19+T cells were risk factors for viral pneumonia complicated with ARDS(P<0.05).Conclusion:Serum NLR,IL-6,CD3+T and CD19+T cells are closely related to severity and prognosis of patients with viral pneumonia complicated with ARDS.

Objective To compare the effects of cortical bone trajectory ( CBT) and traditional trajectory ( TT)pedicle screw internal fixation on the range of motion (ROM) and rod system stress of normal and osteoporotic(OP) spines. Methods The L3-S1 finite element models of normal and OP spines were established. The screwrod system with two kinds of trajectory was used for internal fixation of the L4-5 segment, so as to simulate sixphysiological loads, namely, flexion, extension, left / right bending, left / right rotation. The effects of two internalfixation methods on ROMs and maximum equivalent stress of screws in normal and OP spines were compared.Results For both bone conditions, CBT and TT significantly reduced ROM of the fixed segment (L4-5) and theentire segment of lower lumbar spine ( L3-S1). However, the ROM decline of CBT group was slightly smaller than that of TT group, and their ROMs were similar under flexion and extension, but the ROM differences were significant under lateral bending and axial rotation. In addition, for both the normal and OP spine models, themaximum equivalent stress of screws in CBT group was significantly higher than that in TT group. Compared withTT group, the screw stress of CBT group in normal spine model under flexion and extension, lateral bending,axial rotation was increased by 27% , 268% and 58% , respectively. However, when CBT technique was used atthe same time, the OP spine model had a smaller screw stress distribution than the normal spine model.Conclusions Compared with TT technique, CBT technique can achieve higher screw stress under OP conditionand reduce screw stress concentration under normal bone condition. In addition, CBT slightly increases ROMs of each segment, which is conducive to recovery of spinal physiological function after surgery. Lateral bending and axial rotation can produce negative mechanical effects, and these two physiological loads should be avoided.

With the rapid development of information technology and the change in military education policy in the new era, online teaching has gradually become one of the main approaches to implement the clinical medical education in military teaching hospitals. In this study, the online teaching was performed mainly by pre-recorded teaching, supplemented by online live teaching. Massive open online courses (MOOCs) and the seminar-style teaching were advocated to be used in advanced disciplines. The quality of online teaching was guaranteed through infrastructure provision, teacher arrangement, teaching preparation, teaching interactivity, after-class test, teaching evaluation, supervision, and summary of teaching, which ultimately achieved a good effect. The results of the two-way questionnaires of 26 teachers and 129 students showed that 23.26% (30/129) of students and 65.38% (17/26) of teachers believed that students' learning ability was insufficient. 34.88% (45/129) of students and 23.08% (6/26) of teachers thought that the existing technology could not meet the requirements of online teaching. 55.04% (71/129) of students and 69.23% (18/26) of teachers held the view that the effect of online teaching was inferior to face-to-face teaching. 28.68% (37/129) of students and 57.69% (15/26) of faculty asked for the return of face-to-face education. Therefore, it is necessary to strengthen the construction of smart classroom platforms, establish a well-developed online teaching quality evaluation system, and integrate various innovative teaching modes with the online teaching. In these ways, it is expected to optimize the online teaching of clinical medicine and achieve the goal of online teaching.

We conducted a randomized, open-label, parallel-controlled, multicenter trial on the use of Shuanghuanglian (SHL), a traditional Chinese patent medicine, in treating cases of COVID-19. A total of 176 patients received SHL by three doses (56 in low dose, 61 in middle dose, and 59 in high dose) in addition to standard care. The control group was composed of 59 patients who received standard therapy alone. Treatment with SHL was not associated with a difference from standard care in the time to disease recovery. Patients with 14-day SHL treatment had significantly higher rate in negative conversion of SARS-CoV-2 in nucleic acid swab tests than the patients from the control group (93.4% vs. 73.9%, P = 0.006). Analysis of chest computed tomography images showed that treatment with high-dose SHL significantly promoted absorption of inflammatory focus of pneumonia, which was evaluated by density reduction of inflammatory focus from baseline, at day 7 (mean difference (95% CI), -46.39 (-86.83 to -5.94) HU; P = 0.025) and day 14 (mean difference (95% CI), -74.21 (-133.35 to -15.08) HU; P = 0.014). No serious adverse events occurred in the SHL groups. This study illustrated that SHL in combination with standard care was safe and partially effective for the treatment of COVID-19.
COVID-19 ; Humans ; Medicine, Chinese Traditional ; Research ; SARS-CoV-2 ; Treatment Outcome

Objective:To evaluate the clinical efficacy and safety of using the F4.8 Visual Puncture Micro-percutaneous nephrolithotomy to treat the female distal calculi associated with stricture.Methods:From June 2017 to December 2019, 32 female patients with distal ureteric calculi associated with stricture, aged(35.0±10.3) years ( range from 16 to 75 years)old, were enrolled into this retrospective study. They were diagnosed by colour doppler ultrasound, IVU(intravenous Urography), or CT, et al. The average stone size was (13.0±3.6)mm in diameter(range from 3 to 20 mm), and the stone obstruction duration was from 2 to 35 days, with average of(5±17) days. Twenty-seven cases were on the unilateral ureter and 5 cases were on the bilateral ureters. There were 17 cases undergoing ESWL 2 weeks before. Six cases of stone diameter less than 6 mm were administered medical therapy for more than 7 days. All the 32 case underwent ureteroscopic lithotripsy but failed because of the ureter stricture. They all suffered from hydronephrosis, with the diameter of renal collecting system from 15 to 45 mm, with(23±15)mm on average. The lithotomy position was taken, and the F4.8 Visual Micro-percutaneous nephrolithotomy using 0.9% Sodium chloride was applied to enter into ureter through urethra. There were 21 cases of ureter orifice stricture, including 8 cases associated with avulsion or perforation, 9 cases associated with intramural ureter abnormality and stricture, the zebra guidewire being failed to enter. The F4.8 Visual Micro-percutaneous nephrolithotomy cooperated with water pressure modulation was used to flush and open the ureter orifice and intramural ureter for entering. Holmium lase was used for lithotripsy. Two cases stone were infective and obstructed seriously. F4.8 Visual Micro-percutaneous nephrolithotriptor entered the ureter and destroyed the stones, and the stone fragments were discharged. Stone migrated upward and escaped occurred in 1 case, then the zebra guidewire was indwelled and ureteroscope was used for lithotripsy successfully. All cases were indewelled F4.7 doubld-J tube and urethral catheter after operation.Result:All the 32 patients underwent lithotripsy successful by one-stage. The operation time was 15-43 min [averaged (35.0±8.7)min]. All patients were recovered and discharged 4-7(averaged 5.3) days after operation. Three patients occured fever, T>38.5℃, and they recovered by using sensitive antibiotics according to the blood and urine culture. No severe complications occurred, such as ureteral perforation or extravasation. All patients were reviewed by ultrasound and KUB 3 days after operation, finding 24 cases with stone free, and 8 cases of a little residual stone, with the stone free rate of 75%. One month later, ultrasound and IVU was performed, and no residual stone was detected, with the stone free rate of 100%. The hydronephrosis alleviated by varying degrees. The diameter of the renal collecting system was from 0 to 35 mm, with(12±9)mm on average. The IVU showed the ureter was unobstructed. Conclusions:The F4.8 Visual Micro-percutaneous nephrolithotomy is safe and effective for the female distal calculi associated with stricture, when routine ureteroscopic lithotripsy failed.

Objective: To analyze the mutation of target genes in extranodal natural killer/T-cell lymphoma (ENKTL) by using nextgeneration sequencing, and to explore its relationship with prognosis and clinical characteristics, as to provide evidence for the pathogenesis, clinical diagnosis and targeted therapy of ENKTL. Methods: According to previous literature reports, the genes whose mutations can affect the development of lymphoma were selected as the target genes for this study. 29 patients with ENKTL, who were newly diagnosed at the Fourth Hospital of Hebei Medical University from August 2010 to October 2018, were selected for this study. The mutation of 9 target genes in the specimen was detected by thenext-generationsequencingtechnology.Therelationshipsamongclinicalfeatures,diseaseprognosisandmutationofthetargetgeneswereanalyzedbySPSS21.0statisticalsoftware.Results： ：Ninetargetgenes were were screened. AT-rich interactive-domain 1A(ARID1A) gene showed the highest mutation rate in ENKTL (10 cases, 34.48%) followedbylysinemethyltransferase2D(KMT2D)gene(31.03%)andtumorprotein P53 (TP53) gene (24.13%). Kaplan-Meier survival analysis showed that the overall survival of ENKTL patients with KMT2D gene wild type was significantly better than patients with KMT2D gene mutation (P=0.006). The KMT2D gene mutation was found to besignificantlyrelatedtoclinicalstage,CRP,albumin,lymphocyte count and Ki67 expression in ENKTL patients (all P<0.05). COX regression analysis showed that KMT2D gene mutation was an independent adverse prognostic factor (P<0.05). Conclusion: The KMT2D gene has a high mutant frequency in ENKTL and is associated with patients’prognosis, suggesting that KMT2D gene plays an important role in the initiation and development of ENKTL. It could be used as a clinical therapeutic target for ENKTL.

Objective To investigate the expression of melanoma antigen- encoding gene (MAGE) A1 protein in esophageal squamous cell carcinoma, and explore its correlation with the clinicopathological factors and prognosis. Methods A retrospective analysis was performed on 197 patients with esophageal squamous cell carcinoma who accepted radical surgical treatment from January 2006 to December 2012. The expressions of MAGEA1 protein in these specimens of cancer tissue and cancer adjacent tissue were detected by immunohistochemistry with tissue microarray technology. Results MAGEA1 protein was expressed in cytoplasm and nucleus of tumor cells. The positive expression rate of MAGEA1 protein in cancer tissue was significantly higher than that in cancer adjacent tissue: 73.6％ (145/197) vs. 5.6％ (11/197), and there was statistical difference (P<0.01). The positive expression of MAGEA1 protein had no correlations with sex, age, history of smoking/drinking, family history of upper gastrointestinal cancer, depth of tumor invasion, lymph node metastasis, tumor differentiation, location and TNM stage (P>0.05). Kaplan-Meier survival analysis result showed that the 5-year survival rate in patients with MAGEA1 protein positive expression was significantly lower than that in patients with MAGEA1 protein negative expression (37.2％ vs. 53.8％), and there was statistical difference (P=0.018). Multivariate analysis result showed that MAGEA1 protein positive expression was an independent predictor of prognosis in esophageal squamous cell carcinoma patients (HR=1.91, 95％CI 1.22 to 2.98, P = 0.004). Conclusions The expression of MAGEA1 protein is abundant in esophageal squamous cell carcinoma, and is related to worse clinical outcome. MAGEA1 protein could be a candidate target for tumor immunotherapy.

Objective: To investigate the therapeutic outcome of expanded scalp flaps pedicled with superficial temporal vessel for the reconstruction of large facial defects. Method: From Dec 2014 to Oct 2016, 10 cases with large facial skin defects were treated with expanded scalp flaps pedicled with superficial temporal vessel and delayed laser hair removal.Extra expanded scalp flaps were collected as experimental groups. Normal skin(forehead, temporal scalp, cheek, upper eyelid, lower eyelid and nasal dorsum)of 10 cases were collected for control, to compare skin thickness.All patients were followed at least 6 months. Results: There was no significant difference of skin thickness between the expanded scalp flaps and cheek, forehead, nasal dorsum skin(P>0.05). But upper eyelid and lower eyelids skin was significantly thinner than other local skin tissuein controls, and expanded scalp flap (P<0.05). The expanded scalp flap matched well with surrounding tissues in color, texture and thickness. Conclusions It is a good option to repair large facial skin defects with expanded scalp flaps, pedicled with superficial temporal vessel, and laser hair removal, though its shortcoming in eyelid skin defect repairment.

Objective To understand the frequency distribution of CYP2C9 and VKORC1 gene single nucleotide polymor-phisms in Yunnan Han population.Methods CYP2C9(430C> T,1075A>C and 1080C> G)locus and VKORC1(-1639G> A and 1173C> T)locus gene polymorphisms in 202 samples were detected by adopting electrochemical gene sensor method,and the allele frequencies and genotype frequencies were performed the statistics and the gene polymorphism in relevant populations was an-alyzed.Results Among 202 samples,202 cases(100.0%)were genotype C/C at CYP2C9 * 2 locus,C allele frequency was 100.0%;185 cases(91.6%)were genotype A/A at CYP2C9*3 locus,15 cases(7.4%)were A/C genotype,2 cases(1.0%)were C/C genotype,A allele frequency was 95.3%,C allele frequency was 4.7%;CYP2C9*5 locus genotype C/C was in 202 cases (100.0%),C allele frequency was 100%;VKORC1 -1639G > A locus genotype A/A was in 145 cases(71.8%),57 cases (28.2%)were G/A genotype,A allele frequency was 85.9%,G was 14.1%;1173C> T locus genotype T/T was in 145 cases (71.8%),C/T gene type in 57 cases(28.2%),T allele frequency was 85.9%,and C was 14.1%.Conclusion The distribution of CYP2C9 gene in Yunnan Han population is similar to that in other regions of our country.The VKORC1 gene is different from the foreign population,Chinese Han nationality and partial minority nationalities.