Aim To establish a rapid method to efficiently iso-late mononuclear cells from central nervous system ( CNS) tis-sues of mice that can be effectively utilized for identification of various immune cell populations in a single sample by flow cy-tometry. Methods For defining the feasibility and practicality of the method, wild-type C57BL/6 mice and two mouse models of CNS disease including EAE mice and APP/PS1 mice were used in this study. After the collection and homogenization of the brain and spinal cord tissues respectively, the mononuclear cells were isolated by spinning the 70% -30% Percoll gradients. Cell activities were detected by trypan blue staining, and the im-mune population that infiltrated CNS was identified by flow cy-tometry. Results The results of trypan blue staining showed that the survival rate of the isolated cells was above 90% in all groups. Flow cytometry analysis showed that the relative num- bers of lymphocytes infiltrating CNS of EAE and AD mice in-creased significantly compared with wild-type C57BL/6 mice. In addition, the relative numbers of Th1 and Th17 cell subsets me-diating the inflammatory response also increased significantly, while the decreased regulatory T cells frequency was observed in the two mouse models of CNS disease. Conclusions The cells isolated by the 70% ~30% Percoll gradients centrifugation can be effectively utilized for the identification of various immune cell populations in a single sample by flow cytometry. The meth-od described in this article is simple and rapid in operation and with high survival rate and activity of the cells, which can be ap-plied to the study of the mononuclear cells in CNS.

Objective: To elucidate the clinicopathologic characteristics of atypical epithelioid trophoblastic lesions with cyst and fistula formation after cesarean section. Methods: The clinical and pathological data of 4 cases of post-cesarean atypical epithelioid trophoblastic lesions with cyst and fistula formation diagnosed at Women′s Hospital, School of Medicine, Zhejiang University during April 2007 to June 2018 were evaluated by hematoxylin and eosin stain and EnVision two-step immunohistochemical staining technique. Results: The age of the 4 patients ranged from 32 to 41 years, with a mean age of 36.5 years. Three patients recieved cystectomy and one underwent subtotal hysterectomy. Histologically, the lesions were well circumscribed and consisted of uniform cells of medium size, irregularly enlarged with hyperchromatic nuclei and 1 to 2 inconspicuous nucleoli embedded in abundant hyalinized matrix with fibrinoid material in the center. The cells exhibited immunohistochemical feature of chorionic-type intermediate trophoblastic cells (CK18+, p63+ and CD146-). All patients were alive without recurrence during follow-up of 1 to 40 months (mean=22 months). Conclusion Atypical epithelioid trophoblastic lesion with cyst and fistula formation after cesarean section has unique histological features, and its biological behavior and prognosis are still unclear, which need further exploration.

Objective:To investigate the relationship between adiponectin (ADIPOQ) gene polymorphism and postpartum type 2 diabetes mellitus (T2DM) in pregnant women with gestational diabetes mellitus (GDM).Methods:A retrospective study was conducted on 236 GDM postpartum women admitted to the Affiliated Hospital of Jining Medical College from June 2020 to June 2021 as observation subjects. They were divided into a T2DM group and a non T2DM group based on the occurrence of T2DM after delivery. The clinical data of the two groups were compared. The double deoxygenation end termination method was used to detect the single nucleotide polymorphism (SNP) of the ADIPOQ gene, and the four loci rs17366568, rs822395, rs1501299, and rs2241766 were classified. The relationship between ADIPOQ genotype polymorphism and postpartum T2DM was analyzed using a logistic regression model.Results:The G allele carrying the rs2241766 locus in ADIPOQ gene was negatively correlated with the occurrence of T2DM ( OR=0.71, 0.68, P<0.05). Compared with T2DM patients with TT genotype, the GT+ GG genotype at the rs2241766 locus had a lower risk of occurrence for gestational age ≥2 and HbA 1c>85%. Similarly, T2DM patients with pre pregnancy body mass index (BMI)>25 kg/m 2 were more likely to be carriers of the rs2241766 TT genotype ( P=0.026). The (GT+ TT) genotype carrying the T allele at the rs1501299 locus was a protective factor for gestational age and HbA 1c in T2DM patients. Conclusions:The rs2241766 and rs1501299 polymorphisms of the ADIPOQ gene are associated with susceptibility to postpartum T2DM in GDM women. Individuals with rs2241766 and rs1501299 mutant genotypes belong to the high-risk population for T2DM.

Objective To explore the clinical value of micro ribonucleic acid(miR)-27a and miR-1299 in predicting the long-term prognosis of patients with esophageal cancer.Methods A total of 89 patients with esophageal cancer in the hospital were enrolled between June 2020 and June 2022,and all underwent Minimally invasive thoracic and laparoscopic resection of esophageal cancer.The expression levels of miR-27a and miR-1299 were detected by real-time fluorescence quantitative polymerase chain reaction(PCR)before surgery,and level of serum ferritin(SF)was detected by enzyme linked immunosorbent assay.The relationship between miR-27a,miR-1299,SF and clinicopathology in patients with esophageal cancer was analyzed.All were followed up till February 2023 after surgery,and survival of patients was record.The relationship between miR-27a,miR-1299,SF and prognosis was analyzed.The risk factors of long-term prognosis in patients with esophageal cancer were analyzed by multivariate Cox regression model.The relationship between miR-27a,miR-1299 and serum SF was analyzed by Pearson.Results There were significant differences in miR-27a,miR-1299 and serum SF among patients with different tumor staging,lymph node metastasis and differentiation degree(P＜0.05).The results of survival analysis showed that overall survival rates in high-expression miR-27a and SF groups were lower than those in low-expression miR-27a and SF groups(P＜0.05),while which was higher in high-expression miR-1299 group than low-expression miR-1299 group(P＜0.05).Cox regression analysis showed that miR-27a ≥2.54,miR-1299＜4.18 and serum SF ≥223.78 μg/L(P＜0.05)were independent risk factors of postoperative long-term prognosis of patients with esophageal cancer.Pearson correlation analysis showed that level of serum SF was positively correlated with relative expression level of miR-27a(P＜0.05,r=0.612),while negatively correlated with relative expression level of miR-1299(P＜0.05,r=-0.517).Conclusion The miR-27a,miR-1299 and serum SF are related to tumor staging,lymph node metastasis and differentiation degree in patients with esophageal cancer,which can be applied to evaluate postoperative long-term prognosis.

DNA Polymerase gamma ; DNA-Directed DNA Polymerase ; genetics ; Diffuse Cerebral Sclerosis of Schilder ; diagnosis ; drug therapy ; etiology ; genetics ; Heterozygote ; Humans ; Infant ; Male ; Mutation

Objective To analyze the epidemiological characteristics of patients with sea lice injury in coastal baths in Qinhuangdao City, and provide scientific evidence for the importance of prevention of sea lice injury and the early warning of sea lice outbreak. Methods The data of 2 659 patients with sea lice injury, including gender, age composition, severity of bruises and other epidemiological data reported by the coastal bathing stations in Qinhuangdao City from 2016 to 2018 (mainly from July to August) were retrospectively analyzed, and the correlation between the temperature at the surface of sea water and the number of patients with sea lice injury was also analyzed. Results Among the 7 bathing beaches along the coast of Qinhuangdao City, Dongshan Bathing Spot (1 101 cases) and Repulse Bay Bathing Beach (1 365 cases) had the largest numbers of bruises, while the Gold Coast Bath had the least (only 5 cases), and the differences in number of bruises in different baths were statistically significant in 2016, 2017, 2018 years (277, 1 890, 492 cases respectively, χ2= 490.611, P < 0.05). Among 2 659 patients, 1 124 were children (≤14 years old), 1 535 were adult, 1 570 male and 1 089 female patients. The severity type of patients' sea lice injury in each bath was mainly mild to moderate type; among the total injury cases, there were 2 589 cases with mild type injury accounting for 97.37%, 61 cases with moderate type (2.29%) and 9 cases with severe type (0.34%). A linear trend between sea surface temperature and the patients' number of bruises was seen, and there was a positive correlation between the sea surface temperature and the patients' number (r = 0.433, P < 0.05). Conclusion The sea lice injuries in Qinhuangdao City are mainly distributed in the Dongshan and Repulse Bay Baths, the main type of injury is light one, and in the mean time the injury is influenced by factors such as popularization of science, medical care, environmental factors, and collective sea lice drift, etc.

Background:The purpose of this study was to analyze cases of AO31-A2 intertrochanteric fractures (ITFs) and to identify the relationship between the loss of the posteromedial support and implant failure.Methods:Three hundred ninety-four patients who underwent operative treatment for ITF from January 2003 to December 2017 were enrolled.Focusing on posteromedial support,the A2 ITFs were divided into two groups,namely,those with (Group A,n =153) or without (Group B,n =241) posteromedial support post-operatively,and the failure rates were compared.Based on the final outcomes (failed or not),we allocated all of the patients into two groups:failed (Group C,n =66) and normal (Group D,n =328).We separately analyzed each dataset to identify the factors that exhibited statistically significant differences between the groups,In addition,a logistic regression was conducted to identify whether the loss of posteromedial support of A2 ITFs was an independent risk factor for fixation failure.The basic factors were age,sex,American Society of Anesthesiologists (ASA) score,side of affected limb,fixation method (intramedullary or extramedullary),time from injury to operation,blood loss,operative time and length of stay.Results:The failure rate of group B (58,24.07％) was significantly higher than that of group A (8,5.23％) (x2 =23.814,P ＜ 0.001).Regarding Groups C and D,the comparisons of the fixation method (P =0.005),operative time (P =0.001),blood loss (P =0.002)and length of stay (P =0.033) showed that the differences were significant.The logistic regression revealed that the loss of posteromedial support was an independent risk factor for implant failure (OR =5.986,95％ CI:2.667-13.432) (P ＜ 0.001).Conclusions:For AO31-A2 ITFs,the loss of posteromedial support was an independent risk factor for fixation failure.Therefore,posteromedial wall reconstruction might be necessary for the effective treatment of A2 fractures that lose posteromedial support.

Objective:To build a systematic,uninterrupted and all-round continuous collaborative protection strategy for network security control of gene sequencing equipment.Methods:The network transmission,electronic interface,data system and other parts were incorporated into the comprehensive consideration of network security control of gene sequencing equipment.Based on the"overall network security",the risk points that manufacturers of genetic sequencing equipment and institutions using the testing need to pay attention to and control at this stage were proposed from seven aspects including product data architecture,security capability system construction,residual risk maintenance plan,malicious attack defense,vulnerability assessment,cyber security of off-the-shelf software,and remote operation and maintenance support,and further expanded to the whole lifecycle network security control of gene sequencing equipment.Results:By sorting out the risk points of gene sequencing,a systematic,uninterrupted and all-round continuous collaborative protection strategy of network security control of gene sequencing equipment was constructed.Conclusion:Network security supervision of gene sequencing equipment is an important tool to ensure product quality.It is necessary to pay attention to comprehensive risk points in product development,production and maintenance,and network security control should be implemented throughout the whole life cycle of gene sequencing equipment.

Objective To compare the clinical outcomes of using elastic intramedullary nail and plate to fix fibular frac-ture.Methods The 60 patients with tibiofibular fractures admitted from January 2015 to December 2022 were divided into two groups:intramedullary nail group and plate group,30 cases each,intramedullary nail group was treated with elastic in-tramedullary nail fixation group,plate group was treated with steel plate and screw fixation group.Intramedullary nail group,there were 18 males and 12 females,aged from 22 to 75 years old with an average of(39.4±9.8)years old,including 24 cases of traffic accidents injury,6 cases of falling injury,23 cases of closed fractures,7 cases of open fractures.Steel plate group,there were 15 males and 15 females,aged from 24 to 78 years old with an average of(38.6±10.2)years old.The 22 cases were injured by traffic accident,8 cases were injured by falling.The 24 cases were closed fractures and 6 cases were open fractures.The operation time,intraoperative bleeding,American Orthopedic Foot and Ankle Society(AOFAS)ankle and hind foot scores,clinical healing time of fibula and the incidence of wound complications were compared between the two groups.Re-sults The patients in both groups were followed up for 6 to 21 months,with an average of(14.0±2.8)months.Compared with plate group,intramedullary nail group had shorter operative time,less bleeding,shorter clinical healing time of fibula,and low-er infection rate of incision,and the difference was statistically significant(P＜0.05).There were 2 cases of delayed healing in intramedullary nail group,1 case of nonunion in plate group,and 2 cases of delayed healing in plate group,and there was no statistically significant difference between the two groups(P＞0.05).In the last follow-up,according to the AOFAS scoring standard,the ankle function in intramedullary nail group was excellent in 17 cases,good in 12 cases,fair in 1 case,with an av-erage of(88.33±4.57)points,while in plate group,excellent in 16 cases,good in 10 cases,fair in 4 cases,with an average of(87.00±4.14)points;There was no statistical difference between the two groups(P＞0.05).Conclusion Elastic intramedullary nail has the advantages of short operation time,less intraoperative bleeding,short fracture healing time and less incision com-plications in the treatment of fibular fracture,which is worthy of clinical application.

Objective:To investigate the clinical features and influencing factors of liver function injury in patients with 2019-nCoV/SARS-CoV-2 Omicron mutant strains.Methods:1 183 confirmed imported cases of SARS-CoV-2 who were admitted at Shanghai Public Health Clinical Center (affiliated to Fudan University) from July 1, 2021 to January 15, 2022 were collected. Clinical data, viral genotyping and laboratory test results were collected to retrospectively analyze the basic condition and clinical characteristics of liver function injury. Statistical analysis was performed using t-test or Wilcoxon rank-sum test, χ2 test or Fisher's exact test, Pearson correlation test and logistic regression analysis. Results:125 (10.6%) cases had raised baseline ALT level and 60 (5.1%) cases had abnormal baseline AST level. Among them, 33 cases (2.8%) had received hepatoprotective drugs. Liver function injury was generally mild in SARS-CoV-2 infection and minimal in Omicron mutant strains. Leukocyte count was increased in patients with raised alanine aminotransferase (ALT) [(6.96±1.78)×10 9/L vs. (6.41±1.96)×10 9/L, P=0.005 2], CT scan showed the proportion of liver hypodensity was significantly increased (2.4% vs. 0.3%, P=0.018 0). High-sensitivity C-reactive protein [(7.83±22.36) mg/L vs. (2.68±6.21) mg/L, P=0.007 8] and D-dimer [(0.34±0.39) μg/ml vs. (0.31±0.75) μg/ml, P=0.047 5] levels were higher in patients with raised AST than normal group. 26 cases had normal liver function at hospital admission; however, abnormal liver function was occurred during the course of the disease. Another 8 patients had abnormal liver function at hospital admission, and reduced liver function further during the course of treatment. Recovery time and length of hospital stay was significantly affected in patients with worsened liver function. Baseline body mass index value [odds ratio ( OR)]=1.80, P=0.047), non-Omicron strains ( OR=12.63, P=0.046), D-dimer ( OR=2.36, P=0.047) and interleukin-6 levels ( OR=1.03, P=0.009), and those who used glucocorticoids and/or ulinastatin after hospital admission ( OR=6.89, P=0.034) had a higher risk of worsening liver function. Conclusions:Liver dysfunction could be observed among COVID-19 patients. Patients infected with omicron variant generally showed mild liver injury. Dynamic monitoring of liver function is necessary, especially among those with baseline elevated IL-6, D-Dimer level and use of antiinflammation medication during treatment.