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 explore the effects of the antihypertensive drug Amlodipine on calcium influx and autoph- agy in human podocytes ( HPC) ． Methods : HPC cells were routinely cultured in vitro． HPC cells were treated with angiotensin Ⅱ ( Ang Ⅱ ) ，the L-type Ca2 + blocker Amlodipine alone or in combination．The Ca2 + imaging system was used to detect the transient changes in the intracellular Ca2 + flux of HPC cells in real time after drug treatment．Western blot was employed to detect the changes in the ratio of autophagy marker proteins LC3B-Ⅱ/ LC3B-Ⅰ , and the expression levels of Beclin-1，P62，as well as apoptosis-related proteins Bcl-2 and Bax．Flow cytometry was used to detect the number of Fluo-4AM positive cells at 488 nm to analyze the level of intracellular Ca2 + influx in HPC cells．Lyso-Tracker Green live cell staining was applied to analyze the fluorescence intensity of lysosomes．Flow cytometry was also used to detect the apoptosis rate of HPC cells． Results : Compared with the control group，in the Ang Ⅱ group，the transient Ca2 + flux and the number of Fluo-4AM positive cells increased significantly (P＜0. 001) ．The ratio of autophagy marker proteins LC3B-Ⅱ/ LC3B-Ⅰ (P＜0. 001) and the pro- tein expression of Beclin-1 (P＜0. 01) decreased significantly，while the expression of P62 increased (P＜0. 01) ． The fluorescence intensity of lysosomes weakened (P＜0. 05) ，the apoptosis rate increased (P＜0. 0001) ，the ex- pression of apoptosis-related protein Bcl-2 decreased (P ＜0. 01 ) ，and the protein level of Bax increased (P < 0. 001) ．Compared with the control group，in the Amlodipine group，the number of Fluo-4AM positive cells de- creased significantly (P＜0. 001) ，the ratio of LC3B-Ⅱ/ LC3B-Ⅰ (P＜0. 001) and the protein expression of Bec- lin-1 (P＜0. 001) increased，the protein expression of P62 decreased (P＜0. 05) ，the fluorescence intensity of ly- sosomes enhanced (P＜0. 01) ，the apoptosis rate decreased (P＜0. 01) ，the protein expression of Bcl-2 increased (P＜0. 001) ，and the protein level of Bax decreased (P＜0. 001) ．Compared with the Ang Ⅱ group，in the Ang Ⅱ + Amlodipine group，the number of Fluo-4AM positive cells decreased significantly (P＜0. 001) ，the ratio of LC3B-Ⅱ/ LC3B-Ⅰ (P＜0. 01) and the protein expression of Beclin-1 increased (P＜0. 05) ，the protein level of P62 decreased (P＜0. 01) ，the fluorescence intensity of lysosomes increased (P ＜0. 05) ，the apoptosis rate de- creased (P＜0. 001) ，the protein expression of Bcl-2 increased (P ＜0. 001 ) ，and the protein level of Bax de- creased (P＜0. 001) ． Conclusion Amlodipine inhibits calcium influx，promotes autophagy and inhibits apoptosis in human podocytes，which is useful in preventing the development of hypertensive nephropathy．

Objective To investigate the effects of traditional laparoscopic surgery,natural orifice specimen extraction surgery(NOSES),and intersphincteric resection(ISR)on treatment outcomes and quality of life in patients with low rectal cancer.Methods A total of 152 patients with low rectal cancer who were admitted from January 2020 to June 2022,and they were divided into the traditional laparoscopic group(49 cases),the NOSES group(51 cases),and the ISR group(52 cases)according to the surgical method.The operation status,postoperative recovery status,pain,anal function recovery status,quality of life and complications were compared in the 3 groups.Results The operation time of the traditional laparoscopic group[(193.98±12.31)min]was lower than that of the NOSES group[(203.54±15.02)min]and the ISR group[(199.85±11.98)min](P＜0.05),operation time of NOSES group and ISR group was no difference(P＞0.05).The first exhaust time[(60.21±10.05)h],the first time of getting out of bed[(37.52±6.21)h],and the length of postoperative hospital stay[(12.51±1.47)d]in the traditional laparoscopic group were all higher than those in the NOSES group[(51.06±8.67)h,(30.13±4.92)h,and(11.27±)1.23)d]and ISR group[(53.19±9.24)h,(28.97±4.71)h,(11.73±1.35)d](P＜0.05),and there were no statistically significant differences in the first exhaust time,the first time to get out of bed,and the length of postoperative hospital stay between the NOSES and ISR groups(P＞0.05).There was no statistically significant difference in the Visual Analogue Scale(VAS)scores for pain at 4 hours,24 hours,and 48 hours after surgery among the three groups(P＞0.05).The VAS scores of the three groups at 24 hours after surgery were higher than those at 4 hours and 48 hours after surgery,and the difference was statistically significant(P＜0.05).The VAS scores of the three groups at 48 hours after surgery were higher than those at 4 hours after surgery,and the difference was statistically significant(P＜0.05).The NOSES group's Wexner score[(4.93±0.76)points]at 3 months after surgery and Wexner score[(3.21±0.42)points]at 6 months after surgery were lower than those of the ISR group[(6.32±0.93)points,(4.48±0.54)points]and the traditional laparoscopic group[(5.93±0.81)points,(4.01±0.53)points](P＜0.05),and the Wexner score of the 3 groups at 3 months after surgery was lower than that at 1 month after surgery(P＜0.05).The EORTC QLQ-C30 score of the NOSES group at 3 months after surgery was(74.82±4.01)points,and that at 6 months was(85.49±4.93)points,which were higher than those of the ISR group[(67.05±5.03)points and(71.64±4.21)points]and the traditional laparoscopic group[(70.42±3.92)points,(76.28±4.48)points](P＜0.05),and the EORTC QLQ-C30 scores of the traditional laparoscopic group at 3 and 6 months after surgery were higher than those of the ISR group,and the difference was statistically significant(P＜0.05).The EORTC QLQ-C30 score of the 3 groups at 6 months after surgery was higher than that before surgery and 3 months after surgery(P＜0.05),and the EORTC QLQ-C30 score of the 3 groups at 3 months after surgery was higher than that before surgery(P＜0.05).There was no significant difference in the incidence of total complications among the three groups(P＞0.05).Conclusion Compared with traditional laparoscopic surgery for low rectal cancer,the NOSES and ISR methods accelerate postoperative bowel function recovery,and the NOSES methods have advantages in anal function recovery and better and satisfactory quality of life.

Objective To investigate the effects of traditional laparoscopic surgery,natural orifice specimen extraction surgery(NOSES),and intersphincteric resection(ISR)on treatment outcomes and quality of life in patients with low rectal cancer.Methods A total of 152 patients with low rectal cancer who were admitted from January 2020 to June 2022,and they were divided into the traditional laparoscopic group(49 cases),the NOSES group(51 cases),and the ISR group(52 cases)according to the surgical method.The operation status,postoperative recovery status,pain,anal function recovery status,quality of life and complications were compared in the 3 groups.Results The operation time of the traditional laparoscopic group[(193.98±12.31)min]was lower than that of the NOSES group[(203.54±15.02)min]and the ISR group[(199.85±11.98)min](P＜0.05),operation time of NOSES group and ISR group was no difference(P＞0.05).The first exhaust time[(60.21±10.05)h],the first time of getting out of bed[(37.52±6.21)h],and the length of postoperative hospital stay[(12.51±1.47)d]in the traditional laparoscopic group were all higher than those in the NOSES group[(51.06±8.67)h,(30.13±4.92)h,and(11.27±)1.23)d]and ISR group[(53.19±9.24)h,(28.97±4.71)h,(11.73±1.35)d](P＜0.05),and there were no statistically significant differences in the first exhaust time,the first time to get out of bed,and the length of postoperative hospital stay between the NOSES and ISR groups(P＞0.05).There was no statistically significant difference in the Visual Analogue Scale(VAS)scores for pain at 4 hours,24 hours,and 48 hours after surgery among the three groups(P＞0.05).The VAS scores of the three groups at 24 hours after surgery were higher than those at 4 hours and 48 hours after surgery,and the difference was statistically significant(P＜0.05).The VAS scores of the three groups at 48 hours after surgery were higher than those at 4 hours after surgery,and the difference was statistically significant(P＜0.05).The NOSES group's Wexner score[(4.93±0.76)points]at 3 months after surgery and Wexner score[(3.21±0.42)points]at 6 months after surgery were lower than those of the ISR group[(6.32±0.93)points,(4.48±0.54)points]and the traditional laparoscopic group[(5.93±0.81)points,(4.01±0.53)points](P＜0.05),and the Wexner score of the 3 groups at 3 months after surgery was lower than that at 1 month after surgery(P＜0.05).The EORTC QLQ-C30 score of the NOSES group at 3 months after surgery was(74.82±4.01)points,and that at 6 months was(85.49±4.93)points,which were higher than those of the ISR group[(67.05±5.03)points and(71.64±4.21)points]and the traditional laparoscopic group[(70.42±3.92)points,(76.28±4.48)points](P＜0.05),and the EORTC QLQ-C30 scores of the traditional laparoscopic group at 3 and 6 months after surgery were higher than those of the ISR group,and the difference was statistically significant(P＜0.05).The EORTC QLQ-C30 score of the 3 groups at 6 months after surgery was higher than that before surgery and 3 months after surgery(P＜0.05),and the EORTC QLQ-C30 score of the 3 groups at 3 months after surgery was higher than that before surgery(P＜0.05).There was no significant difference in the incidence of total complications among the three groups(P＞0.05).Conclusion Compared with traditional laparoscopic surgery for low rectal cancer,the NOSES and ISR methods accelerate postoperative bowel function recovery,and the NOSES methods have advantages in anal function recovery and better and satisfactory quality of life.

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.

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.

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.

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.

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 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.