【Objective】 To observe the effect of puerarin on the concentration of Ca²⁺ and the expression of brain derived neurotrophic factor (BDNF) in hippocampal neurons of vascular dementia (VD) rats so as to explore the mechanism of puerarin in protecting nerve cells. 【Methods】 Male SD rats were randomly divided into sham operation group, model group, and puerarin intervention group. The vascular dementia model was established by ligating bilateral common carotid arteries at intervals of 3 days. Two weeks after the operation, the learning and memory abilities of the rats were evaluated by Morris water maze, and the expression of BDNF in the hippocampus of the rats was detected by immunohistochemistry and Western blotting. The mean fluorescence intensity was measured by flow cytometry to represent the intracellular free Ca²⁺ concentration. 【Results】 In the puerarin intervention group, the rats’ escape latency in Morris water maze was significantly shortened, the expression of BDNF in the hippocampus was significantly increased, and the concentration of Ca²⁺ in hippocampal neurons was decreased. Compared with the model group, the difference was statistically significant (all P<0.05). 【Conclusion】 Puerarin has neuroprotective effect on VD rats, and its mechanism may be related to the decrease of Ca²⁺ concentration in hippocampal neurons and the up-regulation of BDNF expression.

BACKGROUND: Red-cell transfusion is critical for surgery during the peri-operative period; however, the transfusion threshold remains controversial mainly owing to the diversity among patients. The patient's medical status should be evaluated before making a transfusion decision. Herein, we developed an individualized transfusion strategy using the West-China-Liu's Score based on the physiology of oxygen delivery/consumption balance and designed an open-label, multicenter, randomized clinical trial to verify whether it reduced red cell requirement as compared with that associated with restrictive and liberal strategies safely and effectively, providing valid evidence for peri-operative transfusion. METHODS: Patients aged >14 years undergoing elective non-cardiac surgery with estimated blood loss > 1000 mL or 20% blood volume and hemoglobin concentration <10 g/dL were randomly assigned to an individualized strategy, a restrictive strategy following China's guideline or a liberal strategy with a transfusion threshold of hemoglobin concentration <9.5 g/dL. We evaluated two primary outcomes: the proportion of patients who received red blood cells (superiority test) and a composite of in-hospital complications and all-cause mortality by day 30 (non-inferiority test). RESULTS: We enrolled 1182 patients: 379, 419, and 384 received individualized, restrictive, and liberal strategies, respectively. Approximately 30.6% (116/379) of patients in the individualized strategy received a red-cell transfusion, less than 62.5% (262/419) in the restrictive strategy (absolute risk difference, 31.92%; 97.5% confidence interval [CI]: 24.42-39.42%; odds ratio, 3.78%; 97.5% CI: 2.70-5.30%; P <0.001), and 89.8% (345/384) in the liberal strategy (absolute risk difference, 59.24%; 97.5% CI: 52.91-65.57%; odds ratio, 20.06; 97.5% CI: 12.74-31.57; P <0.001). No statistically significant differences were found in the composite of in-hospital complications and mortality by day 30 among the three strategies. CONCLUSION: The individualized red-cell transfusion strategy using the West-China-Liu's Score reduced red-cell transfusion without increasing in-hospital complications and mortality by day 30 when compared with restrictive and liberal strategies in elective non-cardiac surgeries. TRIAL REGISTRATION ClinicalTrials.gov, NCT01597232.
Humans ; Adult ; Postoperative Complications ; Erythrocyte Transfusion/adverse effects* ; Blood Transfusion ; Hospitals ; Hemoglobins/analysis*

Objective:To compare the efficacy of arthroscopic anterior cruciate ligament reconstruction using tendon autograft with figure-of-four position and traditional knee hyperflexed position for femoral tunnel creation.Methods:A retrospective case series study was conducted to analyze the clinical data of 46 patients with ACL injury admitted to Second Affiliated Hospital of Harbin Medical University from August 2019 to October 2019, including 26 males and 20 females; aged 24-40 years [(31.1±7.5)years]. All patients underwent arthroscopic ACL reconstruction using tendon autograft. The femoral tunnel was created with figure-of-four position in 21 patients (figure-of-four position group) and with traditional knee hyperflexed position in 25 patients (knee hyperflexed position group). The operation time was compared between the two groups. The center position, length and angle of femoral tunnel were evaluated and measured by three dimensional CT reconstruction and Bernard quadrant method at 8 weeks postoperatively. The knee function was assessed by knee Lysholm score preoperatively, at 8 weeks and at 1 year postoperatively. Complications were observed as well.Results:All patients were followed up for 2-20 months [(15.3±2.1)months]. The operation time was (28.5±2.6)minutes in figure-of-four position group, significantly less than (39.5±2.4)minutes in knee hyperflexed position group ( P<0.05). The tunnel center position was located at (27.1±1.4)% and (25.1±2.6)% within the Bernard quadrant in figure-of-four position group, similar with (28.1±2.8)% and (26.1±3.1)% in knee hyperflexed position group (all P>0.05). Total tunnel length and thick tunnel length were (42.1±2.4)mm and (34.1±2.4)mm in figure-of-four position group, significantly longer than (38.2±2.5)mm and (31.1±2.7)mm in knee hyperflexed position group (all P<0.05). The coronal plane angle of the tunnel was (41.1±2.4)° in figure-of-four position group, significantly smaller than (47.5±2.6)° in knee hyperflexed position group ( P<0.05). The sagittal plane angle of the tunnel was (42.1±1.4)° in figure-of-four position group, significantly greater than (37.1±1.8)° in knee hyperflexed position group ( P<0.05). Figure-of-four position group showed the knee Lysholm score of (53.4±5.2)points preoperatively, (97.1±1.4)points at 8 weeks postoperatively and (98.3±2.3)points at 1 year postoperatively. Knee hyperflexed position group showed the knee Lysholm score of (54.3±7.4)points preoperatively, (97.1±1.6)points at 8 weeks postoperatively and (98.1±1.3)points at 1 year postoperatively. The knee Lysholm score did not differ significantly between the two groups (all P>0.05), but the knee function was significantly improved in both groups when compared with that before the operation (all P<0.05). There were 1 patient with femoral tunnel fracture, one with injury to the medial condylar cartilage and one with injury to the posterior root of lateral meniscus in knee hyperflexed position group, while no above-mentioned complications occurred in figure-of-four position group ( P<0.05). Conclusion:Arthroscopic ACL reconstruction using tendon autograft with femoral tunnel creation through figure-of-four position and traditional knee hyperflexed position can both contribute knee functional recovery, but the figure-of-four position has the advantages of short operation time, accurate tunnel positioning, favorable length and angle of the tunnel and less complications.

OBJECTIVE: To analyze the clinical phenotype and variant of SLC2A1 gene in a Chinese pedigree affected with glucose transporter type 1 deficiency syndrome (GLUT1-DS). METHODS: Clinical data of a child who was treated due to delayed motor and language development and his family members were collected. DNA was extracted from peripheral blood samples and subjected to high-throughput medical exome sequencing. Candidate variant was verified by Sanger sequencing of his parents and sister. The genotype-phenotype correlation was explored. RESULTS: The child, his mother and sister had common manifestations such as delayed mental and motor development, poor exercise tolerance, easy fatigue and paroxysmal dystonia, but the difference was that the child and his mother had microcephaly and seizures, while his sister did not. A heterozygous missense SLC2A1 c.191T>C (p.L64P) variant was identified in all affected members, which was unreported previously. CONCLUSION The missense SLC2A1 c.191T>C (p.L64P) variant probably underlay the disease in the proband and his mother and sister. Variability of the clinical phenotypes has reflected the genetic and phenotypic diversity of GLUT1-DS. Detection of the novel variant has enriched the spectrum of GLUT1-DS mutations.
Carbohydrate Metabolism, Inborn Errors ; China ; Glucose Transporter Type 1/genetics* ; Humans ; Monosaccharide Transport Proteins/deficiency* ; Mutation ; Pedigree ; Phenotype

Objective:To investigate the molecular mechanism of PIEZO1 in promoting the invasion and aggression of glioma via Yes-associated protein (YAP) delivering mechanical signals.Methods:(1) Specimen detection: specimens from 94 patients accepted glioma resection in our hospital from February 2015 to March 2017 were chosen; immunohistochemical staining was used to detect the PIEZO1 expression in these glioma tissues of different grades. (2) Cell experiment: human glioma cell lines U87 and U251 were cultured under different matrix; the expressions of PIEZO1 and YAP were detected by immunofluorescent staining, and the PIEZO1 protein expression was detected by Western blotting. (3) Cell experiment after lentivirus transfection: the U87 and U251 cells were divided into negative control group and sh-PIEZO1 group according to the presence or lack of shRNA lentivirus vector; the role of PIEZO1 in glioma proliferation and invasion was detected by clone formation assay, proliferation and invasion assay, and Western blotting; after PIEZO1 silencing, Western blotting was used to detect the expressions of mechanical signal pathway proteins, YAP, FAK, and β1-integrin; real time-PCR was used to detect the mRNA expressions of CTGF and CYR61, which were downstream target genes of YAP; immunofluorescent staining was used to detect the YAP expression after PIEZO1 silencing. Results:(1) The PIEZO1 expression in glioma specimens increased with glioma grading, and the PIEZO1 expression in IDH wild-type patients was higher than that in IDH mutant patients. (2) The PIEZO1 and YAP expressions increased with the increase of matrix stiffness; as compared with that in the 0.2 kPa group, the PIEZO1 protein expression in stromal cultured cells of 16 and 64 kPa groups was significantly increased ( P<0.05). (3) After PIEZO1 silencing, U87 cells became large and tentacles increased, while U251 cells were mostly with long fusiform; as compared with those in the negative control group, the number of cell clones, and the proliferation rate and invasive cell count at each time point in the sh-PIEZO1 group were significantly smaller/decreased ( P<0.05); as compared with those in the negative control group, the epithelial marker E-cadherin expression was significantly increased, while the expressions of stromal markers Vimentin, Snailin and Slug were significantly decreased in the sh-PIEZO1 group ( P<0.05). Finally, Western blotting and immunofluorescent staining showed that PIEZO1 silence significantly increased the phosphorylated-YAP expression, significantly decreased the YAP nuclear expression, significantly down-regulated the CYR61 and CTGF expressions, and significantly reduced the levels of integrin β1 and phosphorylated-FAK; there were significant differences on these indexes between the negative control group and sh-PIEZO1 group ( P<0.05). Conclusion:PIEZO1 regulates the response of glioma cells to mechanical signal and promotes proliferation and invasion of glioma cells through YAP, which is an attractive therapeutic target for glioma treatment.

【Objective】 To investigate the unqualified rate of anti-HIV detection of blood screening laboratories in Beijing-Tianjin-Hebei region, and explore the differences in anti-HIV detection ability and influencing factors in each laboratory. 【Methods】 Through filling questionnaires via e-mail, the anti-HIV ELISA unqualified rate and confirmed (WB) positive results (data) from January to December 2018 from 15 blood screening laboratories in Beijing-Tianjin-Hebei region were collected. Our laboratory was responsible for data collection and confirmation, and statistics software SPSS22.0 was used for analysis. 【Results】 1) There was a statistically significant difference among the unqualified rate of anti-HIV ELISA(6.77‱~35.71‱) and confirmed positive rate(0.60‱~3.56‱) in 15 blood screening laboratories in Beijing-Tianjin-Hebei region (P<0.05); 2) There were significant differencse among the ELISA unqualified rate and the confirmed positive rate of 8 reagents for anti-HIV detection(P<0.01), and the sensitivity of the 4th generation detection reagent and the imported reagent was higher than that of the 3rd generation reagent and the domestic reagent. The anti-HIV ELISA unqualified rate of R5 was the highest (19.08‱). 3)There were significant differences in the anti-HIV ELISA unqualified rate of R1, R2, R3, R5 and R7 reagents among different blood station laboratories(P<0.05), and there were no significant differences in the anti-HIV ELISA unqualified rate of R4, R6 and R8 reagents among different blood station laboratories(P>0.05). 4)The unqualified rate of anti-HIV ELISA of laboratories using different regents showed significant differences(P<0.05), except H, J, M. The unqualified rate of imported reagent was significantly higher than that of domestic reagents of laboratories using imported and domestic reagents combinations(P<0.05), except O. 62.5% (5/8) laboratories using domestic 3^rd and 4^th generation reagent combination showed significant differences in the unqualified rates among different reagents(P<0.05); 5) The positive rate of single-reagent(62.02%~95.45%)in 15 blood screening laboratories showed significant difference(P<0.001), and A was the lowest (62.02%). 【Conclusion】 The anti-HIV detection ability among 15 blood screening laboratories in Beijing-Tianjin-Hebei region is quite different. The application of different reagents is the main factor for the difference, and other factors such as personnel, instruments and test strategies also has a great impact on the detection of anti-HIV. It is still necessary to promote the process of homogenization of blood testing quality among blood screening laboratories in Beijing-Tianjin-Hebei region.

Objective To study the impact of splenectomy and devascularization on liver function and liver fibrosis in patients with non-cirrhotic portal hypertension (NCPH). Methods The clinical data of patients with NCPH who were treated in Beijing You'an Hospital of Capital Medical University from April 2008 to December 2017 were retrospectively analyzed. The patients were divided into the observation group (n=16 ) and the control group ( n =30 ) according to their treatment methods. The observation group underwent splenectomy combined with devascularization, while the control group received conservative treatment. The changes in liver function, ascites, varicose vein rupture and bleeding before and after treat-ment, and the changes in liver blood flow before and after treatment in the observation group were compared. Results The before and after treatment of the 2 groups in ALT, AST, total bilirubin, albumin showed no statistically significant difference (P＞0. 05). Six months after treatment, the rates of ascites and bleeding were significantly higher than the observation group, (P＜0. 05). Ten patients who had a history of preoper-ative variceal hemorrhage in the observation group did not bleed within 6 months after treatment. Further-more, in the observation group after treatment when compared with the control group, the PC-Ⅲ [(32. 3 ± 12. 1) g/L vs. (56. 7 ± 15. 3)g/L],Ⅵ-C [(46. 6 ± 35. 0) g/L vs. (121. 3 ± 30. 4)g/L], LN [(32. 5 ± 10. 5) g/L vs. (65. 8 ± 11. 1) g/L] were significantly lower than the control group ( P ＜0. 05). The preoperative portal venous blood flow in the observation group was significantly higher than that after surgery [(1 056. 8 ± 679. 8) ml/min vs. (481. 0 ± 227. 6) ml/min, P＜0. 05]. Conclusion Splenectomy and devascularization effectively stopped variceal bleeding and delayed liver fibrosis in NCPH patients without affecting the liver function.

As the key phase for medical students to be doctors,clinical internship plays a vital role in the whole medical education process.Interns in the department of gastrointestinal surgery face series of difficulties,such as wide variety of diseases and emergence of new technology and new concepts.In addition,because of the uneven levels of teachers and low enthusiasm of interns,the result of clinical practice teaching is not satisfactory.Based on the above-mentioned problems,measures have been introduced during the process of clinical practice teaching,including selection of typical diseases and surgical procedures,construction of clinical practice teaching team,adoption of pluralistic teaching methods,reinforcement of participation of interns during the clinical practice.Finally,the quality of teaching in our department has been irmproved significandy.

Objective To investigate the risk factors that may induce pneumothorax resulted from subclavian vein puncture during interventional therapy for arrhythmia.Methods The clinical data of 4 351 patients with heart disease,who were admitted to Wuhan Asia Heart Disease Hospital,China,during the period from January 2010 to January 2014 to receive subclavian vein puncture for the performance of radiofrequency catheter ablation or pacemaker implantation,were retrospectively analyzed.The gender,age,presence of chronic obstructive pulmonary disease (COPD) and the shape of clavicle of patients who developed pneumothorax were analyzed,and their relationships with the occurrence of pneumothorax were evaluated.Results Of the 4 351 patients,47 patients developed pneumothorax(1.08％),their age was 12-72 years old with a mean of (47.1±18.4) years old.Among the 47 patients,37 patients were females (78.7％).The angle between clavicle and sternum ＜60°(oblique clavicle) was seen in 27 patients,coexisting COPD was seen in 2 patients.The occurrence of pneumothorax was strikingly higher in females than that in males (OR=2.7,95％ CI=1.4-5.2).In patients with oblique clavicle the risk of pneumothorax was obviously increased (OR=3.5,95％ CI=1.6-7.9) and in patients with COPD the probability of pneumothorax was remarkably increased (OR=2.3,95％CI=1.2-4.7).No special treatment was employed in 5 pneumothorax patients,and the pneumothorax was absorbed by itself.In 19 patients,the pneumothorax was relieved through thoracic puncture and suction.Closed drainage of thoracic cavity was employed in 23 patients.All patients were discharged from hospital with complete rehabilitation.Conclusion Clinically,pneumothorax has been an important clinical problem in interventional therapy.In female patients and in patients who have oblique clavicle or coexisting COPD,pneumothorax is more likely to occur when they receive subclavian vein puncture.

Targeted point mutagenesis through homologous recombination has been widely used in genetic studies and holds considerable promise for repairing disease-causing mutations in patients. However, problems such as mosaicism and low mutagenesis efficiency continue to pose challenges to clinical application of such approaches. Recently, a base editor (BE) system built on cytidine (C) deaminase and CRISPR/Cas9 technology was developed as an alternative method for targeted point mutagenesis in plant, yeast, and human cells. Base editors convert C in the deamination window to thymidine (T) efficiently, however, it remains unclear whether targeted base editing in mouse embryos is feasible. In this report, we generated a modified high-fidelity version of base editor 2 (HF2-BE2), and investigated its base editing efficacy in mouse embryos. We found that HF2-BE2 could convert C to T efficiently, with up to 100% biallelic mutation efficiency in mouse embryos. Unlike BE3, HF2-BE2 could convert C to T on both the target and non-target strand, expanding the editing scope of base editors. Surprisingly, we found HF2-BE2 could also deaminate C that was proximal to the gRNA-binding region. Taken together, our work demonstrates the feasibility of generating point mutations in mouse by base editing, and underscores the need to carefully optimize base editing systems in order to eliminate proximal-site deamination.
APOBEC-1 Deaminase ; genetics ; metabolism ; Animals ; Bacterial Proteins ; genetics ; metabolism ; Base Sequence ; CRISPR-Associated Protein 9 ; CRISPR-Cas Systems ; Cytidine ; genetics ; metabolism ; Embryo Transfer ; Embryo, Mammalian ; Endonucleases ; genetics ; metabolism ; Gene Editing ; methods ; HEK293 Cells ; High-Throughput Nucleotide Sequencing ; Humans ; Mice ; Mice, Inbred C57BL ; Microinjections ; Plasmids ; chemistry ; metabolism ; Point Mutation ; RNA, Guide ; genetics ; metabolism ; Thymidine ; genetics ; metabolism ; Zygote ; growth & development ; metabolism ; transplantation