[Analysis of a family with 11ß-hydroxylase deficiency due to a mutation in the CYP11B1 gene].

This study reports a family of patients with 11ß-hydroxylase deficiency (11ß-OHD) caused by a novel mutation in the CYP11B1 gene, and analyzes its clinical and genetic characteristics. The clinical data of a patient with intractable hypertension at Air Force Medical Center on May 16, 2014 were retrospectively analyzed. The patient was clinically diagnosed with congenital adrenal cortical hyperplasia. The clinical data of the patient were further collected and the peripheral blood samples of the patient, his parents and his sister were collected for CYP11B1(NM_000497) gene sequencing, suggesting that the patient had compound heterozygous mutations in exon 1:c.199delG, p.Glu67Lysfs*9 and exon 5:c.905_907 delATGinsTT, p.Asp302Valfs*23, both of which were pathogenic variants. The patient's father and sister carried heterozygous mutations in exon 1:c.199delG, p.Glu67Lysfs*9, and the mother carried heterozygous mutations in exon 5:c.905_907delATGinsTT, p.Asp302Valfs*23. This study is the first to report a new compound heterozygous mutation in exon 1:c.199delG and exon 5 c.905_907 delATGinsTT of CYP11B1 gene, enriching the database of 11ß-OHD mutations and providing information to further understand the genetic mechanism of the disease.

[Research progress on the characteristics of gut microbiome in early life and its influence on atopic march].

Allergic diseases are affected by both genetic background and environmental factors.In recent years, many studies have shown that allergic diseases are closely related to the gut microbiome.This article will elaborate on the composition of gut microbiome in early life and its relationship with allergies, the mechanism of action, and the influence of gut microbiome colonization on the atopic march, in order to improve the understanding of the relationship between allergy prevention or treatment and gut microbiome in children, and provide new ideas for the early prevention of allergic diseases and the early intervention of allergic processes.

[Pay attention to the diagnosis and treatment of "easily neglected" complications in liver cirrhosis].

Managing cirrhosis complications is an important measure for improving patients' clinical outcomes. Therefore, in order to provide a complete disease assessment and comprehensive treatment, improve quality of life, and improve the prognosis for patients with cirrhosis, it is necessary to pay attention to complications such as thrombocytopenia and portal vein thrombosis in addition to common or severe complications such as ascites, esophagogastric variceal bleeding, hepatic encephalopathy, and hepatorenal syndrome. The relevant concept that an effective albumin concentration is more helpful in predicting the cirrhosis outcome is gradually being accepted; however, the detection method still needs further standardization and commercialization.

[An analysis of related factors in thrombocytopenia combined with cirrhosis: a cross-sectional study of 2 517 cases].

Objective: To explore the related factors of thrombocytopenia (TCP) occurrence in patients with cirrhosis. Methods: A cross-sectional study was conducted. Inpatients with an initial diagnosis of cirrhosis at Peking University First Hospital from January 1, 2010 to December 31, 2020 were included. Clinical data such as demographic characteristics, etiology of cirrhosis, complications of cirrhosis, laboratory indicators, Child-Pugh grade, invasive procedures, and mortality during hospitalization were collected. A logistic regression model was used to explore the related factors of TCP occurrence in patients with cirrhosis. Categorical variables were compared by the χ(2) test. The inter-group comparison was performed using continuous variables, a t-test, one-way analysis of variance (ANOVA), or a nonparametric test. Results: There were a total of 2 592 cases of cirrhosis. 75 cases with incomplete clinical data were excluded. 2 517 cases were included for analysis. The median age was 58 (50, 67) years. Males accounted for 64%. 1 435 cases (57.0%) developed TCP, and 434 cases (17.2%) had grade 3-4 TCP. Gender, primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), and concomitant esophagogastric varices (EGV) were the major factors associated with TCP. Females were more prone to combine with TCP (OR=1.32, 95%CI: 1.12-1.56, P=0.001). Patients combined with EGV (OR=3.09, 95%CI: 2.63-3.65, P<0.001) were more prone to develop TCP, which was associated with the increased incidence of hypersplenism (P<0.001). Patients with PBC (OR=0.64, 95%CI: 0.50-0.82, P<0.001) and PSC (OR=0.23, 95%CI: 0.06-0.65, P=0.010) were less prone to develop TCP, which was due to the shorter prothrombin time and better coagulation function of PBC patients (P<0.001), and the lower proportion of hypersplenism in combined PSC patients (P=0.004). Patients with TCP and grade 3-4 TCP had a higher rate of hemostatic procedures (P<0.05), but a lower rate of liver biopsy (P<0.05). Patients with grade 3-4 TCP had a higher nosocomial mortality rate compared to those without (P=0.004). Conclusion: TCP is common in patients with cirrhosis. However, TCP occurrence is higher in female patients with EGV and lower in patients combined with PBC and PSC. TCP affects invasive procedures and is associated with adverse outcomes.

[Analysis of influencing factors of recurrence after en bloc spondylectomy of spinal tumors].

Objectives: To investigate the survival and tumor recurrence after en bloc spondylectomy of spinal tumor and analyze the risk factors of postoperative tumor recurrence. Methods: This is a retrospective case series study. Data of 101 patients undergoing en bloc spondylectomy of spinal tumors in the Musculoskeletal Tumor Center, Peking University People's Hospital from December 2006 to June 2022 were analyzed. There were 58 males and 43 females, aged (38.2±15.8) years (range: 10 to 79 years) at the time of surgery; the follow-up time was(44.0±36.0) months (range: 12 to 171 months).Among them, there were 25 relapsed patients, with 7 females and 18 males; aged (34.8±16.3) years (range: 12 to 66 years) at the time of surgery. The types of tumors included 5 giant cell tumors of bone, 6 osteosarcomas, 1 chordoma, 5 chondrosarcomas, 1 undifferentiated sarcoma, 1 fibrosarcoma, 2 Ewing sarcomas, 3 metastases and 1 malignant giant cell tumor of bone. Survival analysis of overall and relapsed patients were performed using the Kaplan-Meier curves. A segmented regression model was used to fit the sequence of recurrence rate changes over time since admission and identify change points for further analysis on risk factors. Univariate and multivariate Logistic regression analysis were performed to assess risk factors associated with recurrence rate; results from multivariate regression analysis were presented using a forest plot. Results: The tumor recurrence rate after en bloc spondylectomy was 24.8% (25/101).The overall median recurrence-free survival after en bloc spondylectomy was 161 months (95%CI: 92 months to NA).The median recurrence-free survival of recurrent patients was 13 months (95%CI: 12 to 27 months).Regarding the classification based on tumor malignancy, and relapse-free survival of metastatic tumors was significantly shorter (P=0.007); and among the surgical margin groups, relapse-free survival of R0 group was significantly better than the R1 and R2 groups (P<0.01). According to the segmented regression model, the tumor recurrence rate for en bloc spondylectomy showed a significant downward trend over time, with relatively higher recurrence rates before 2009 and a relatively stable trend after 2014. The results of univariate analysis showed that surgical margin and time of admission were the influencing factors of patient recurrence. The results of multivariate analysis showed that the R1 resection(OR=13.453,95%CI:2.897 to 97.941,P=0.002) and R2 resection(OR=11.379,95%CI:2.658 to 79.429,P=0.003) were independent influencing factor affecting patient recurrence. Conclusions: The overall tumor recurrence rate after en bloc spondylectomy was high. The surgical margin of tumor resection is an independent risk factor affecting tumor recurrence. Specifically, R2 and R1 resections significantly increase the risk of tumor recurrence.

[Research progress on cirrhosis reversal and recompensation].

Previously, liver lesions in cirrhosis were considered irreversible, especially because the condition aggravated gradually after entering the decompensated phase, thus making it difficult to return to the compensated phase. At present, more and more evidence shows that some patients with decompensated liver cirrhosis can be recompensated after the cause is controlled and complications are managed. This article explores the research progress related to LC reversal and recompensation from three aspects: liver histopathology, liver function, and clinical complications.

[The surgical classification and surgical strategy for en bloc resection of spine tumor].

En bloc resection of spine tumor is the essential treatment method for primary malignant spine tumor,primary aggressive spine tumor and solitary spine metastasis.It can achieve good local control and improve the prognosis.However,given the complex anatomic structure and major adjacent tissue and organ of the spine,en bloc resection of spine tumor remains the research hotspot in surgical treatment of spine tumor.On the basis of adequate surgical classification and surgical strategy,experienced surgical team for spine tumor can achieve good surgical margin while decreasing the morbidity of the en bloc resection of spine tumor.However,the details of surgical classification and surgical strategy require further study.

[Three cases of occupational acute trimethyltin chloride poisoning].

Trimethyltin chloride is a highly toxic substance, which is absorbed through respiratory tract, skin and digestive tract, with central nervous system injury as the main clinical manifestations, and can be accompanied by damage to various organs. In this paper, the treatment process of 3 patients with acute trimethyltin chloride poisoning was reviewed, and their clinical manifestations, auxiliary examination, diagnosis and treatment were analyzed. Three patients were misdiagnosed as mental abnormality, encephalitis, and hepatic encephalopathy in different hospitals in the early stage of medical treatment, suggesting that clinicians should pay attention to the occupational contact history of poisoned patients and conduct toxicant detection in time to avoid misdiagnosis and mistreatment.

[Considerations on expanding the indications for antiviral therapy from the perspective of staging of chronic hepatitis B virus infection].

Patients with chronic (hepatitis B virus,HBV) infection can be divided into immunotolerant, immunoclearance (HBeAg-positive, immune-active), immunocontrol (inactive), and reactivation (HBeAg-negative, immune-active) phases according to HBV serological markers, HBV DNA, alanine aminotransferase, and liver pathology results. Chronic HBV infection is considered indeterminate when the above four phasing criteria are not met. The Chinese "Guidelines" recommend antiviral B treatment for chronic HBV-infected patients with elevated alanine aminotransferase levels after excluding other potential causes. As a result, patients with chronic HBV infection in the immunoclearance and reactivation phases are included in the indication population for antiviral therapy, and the expanded indications are mainly for other infected individuals beyond these two phases: immunotolerant, immunocontrol, and indeterminate. Antiviral therapy may benefit individuals in an indeterminate phase, because they are at a relatively high risk of disease progression.

[Clinical characteristics of 500 hospitalized patients with liver cirrhosis: a retrospective analysis from a tertiary hospital in Beijing].

Objective: To understand the clinical characteristics of hospitalized patients with liver cirrhosis, so as to provide theoretical basis for disease diagnosis and treatment, formulation of intervention measures, and improve the level of disease diagnosis and treatment. Methods: Hospitalized patients who were initially diagnosed with liver cirrhosis at Peking University First Hospital from August 2017 to December 2018 were selected retrospectively as the research objects. Liver cirrhosis demographic data, etiology, severity classification, incidence of complications, diagnosis and prognosis were recorded. Statistical analysis was performed using SPSS software. Results: Among all liver cirrhosis cases, there were 291 males and 209 females, with a male-to-female ratio of 1.4:1 and an age of 59.5±12.9 years as at August 2017 to December 2018. HBV infection, alcoholic liver disease, and autoimmune liver diseases were the most common etiology of liver cirrhosis. HBV infection alone, HBV infection combined with other factors, alcoholic liver disease alone, alcoholic liver disease combined with other factors, autoimmune liver disease alone, and autoimmune liver disease combined with other factors were presented in 163 (32.6%), 57 (11.4%), 47 (9.4%), 63 (12.6%), 85 (17.0%), and 22 (4.4.0%) cases, respectively. Ascites (221 cases, 44.2%), followed by esophagogastric varices (214 cases, 42.8%), and other including hypersplenism (137 cases), liver cancer (126 cases), upper digestive system tract hemorrhage (66 cases), hepatic encephalopathy (40 cases), infection (37 cases), portal vein thrombosis (23 cases), hepatorenal syndrome (20 cases) were the most common complications. The most common site of infection was the abdominal cavity (20 cases), accounting for 54.1%; followed by respiratory tract infection (8 cases), accounting for 21.6% in patients with liver cirrhosis with concurrent infection. Among them, there were 32 cases of bacterial infection alone, one case of bacterial infection combined with fungal infection, one case of bacterial infection combined with viral infection, and three cases of unknown pathogens. There were 69 cases in Child Pugh grade C, and the average hospitalization times were 12.6 days in terms of prognosis. There were total seven cases of death, of which five cases were due to upper gastrointestinal hemorrhage and two due to hepatic encephalopathy. Conclusion: HBV infection, ascites, and upper gastrointestinal bleeding were the most common etiologies, complications, and causes of death in patients with liver cirrhosis at our hospital.

[Effects of visual restoration on dynamic plantar pressure features in elder individuals].

OBJECTIVE: To analyze the effects of visual restoration after cataract surgery on plantar pressure and biomechanics of foot in elder individuals. METHODS: Thirty-two patients [male/female 5/27, (70.1±5.2) years old] with age-related cataract were recruited between October 2016 and December 2019. The footscan system was employed to record the data of plantar pressure during level walking before and 1-month after the cataract surgery. Parameters of peak pressure (PP), impulse (I), pressure-time integral (PTI) and time to peak pressure (TPP) from the regions of the 1st toe (T1), 2nd to 5th toes (T2-5), 1st to 5th metatarsal heads (M1-M5), midfoot (MF), medial hindfoot (HM) and lateral hindfoot (HL) were analyzed respectively. RESULTS: Post-operatively, the visual function was effectively reconstructed with improved visual acuity in both eyes (Z=-4.878, -4.801; P < 0.001). The PP (t=2.266, P=0.031) and I (t=2.152, P=0.039) values in M2 region on the dominant side (right foot) increased statistically at post-operative phase, while the changes of pressure and temporal para-meters in other regions remained stable. There was laterality in plantar pressure at pre-operative phase, manifested as greater PP values in M1, M2, MF, and HM regions on the dominant sides (t=-2.414, -2.478, -2.144, -5.269; P < 0.05), greater PP values in T1, M3, M5 and HL regions on the non-dominant sides (t=4.830, 3.155, 2.686, 3.683; P < 0.05), greater I values in M1, MF, and HM regions on the dominant sides (t=-2.380, -2.185, -5.320; P < 0.05) and greater I values in T1, M3, M5 and HL regions on the non-dominant sides (t=4.489, 2.247, 2.838, 3.992; P < 0.05). post-operatively, the pressure tended to be compatible between the two sides in regions of M3 and MF, while the magnitude of laterality in regions of M1 (ZPP△= -2.721, P=0.007; ZI△=-2.581, P=0.010), M2 (ZPP△=-2.674, P=0.007; ZI△=-2.375, P=0.018) and M5 (ZPP△=1.991, P=0.046; ZI△=2.150, P=0.032) was further increased. CONCLUSION: Changes in plantar pressure after cataract surgery were characterized as increased pressure in the 2nd metatarsal head area on the dominant side. Visual restoration might intensify the laterality in the medial of forefoot on the dominant side and the lateral of forefoot on the non-dominant side.

[Research progress and prospect of liver cirrhosis].

Liver cirrhosis is the final stage of many chronic liver diseases, and is still a heavy disease burden. The proportion of liver cirrhosis caused by the hepatitis B virus is declining, while that caused by the non-alcoholic fatty liver disease (metabolic-associated fatty liver disease) is rising. Several predictive models and techniques such as transient elastography have been used for the early non-invasive evaluation of liver cirrhosis. Effective etiological treatment and complication management are the possible key to reverse and recompense liver function during liver cirrhosis treatment. In recent years, the effectiveness and availability of anti-hepatitis B and C virus drugs have been significantly improved, which provides the basis for effective etiological treatment of liver cirrhosis. However, there is still a lack of etiological treatment measures for non-alcoholic fatty liver disease. Therefore, in addition to focusing on common complications, we should also manage "rare" complications. This article reviews the changes in epidemiological characteristics, the update of the natural history concepts, diagnostic evaluation methods, and the treatment measures for liver cirrhosis.

[Clinical features of Epstein-Barr virus infection associated to liver injury in adolescents and adults].

Epstein-Barr virus (EBV) infection is closely associated to liver injury with diverse clinical features in adolescents and adults. It is often manifested as infectious mononucleosis syndrome, sometimes causing self-limited acute hepatitis, with mild to moderate elevation of liver transaminases, and relative increase in age-related conditions. EBV infection can also cause cholestatic hepatitis, with elevated alkaline phosphatase and γ-glutamyltransferase as the main manifestations, accompanied by varying degrees of jaundice. A small number of patients with severe EBV infection may experience liver failure, and if left untreated in time, it may lead to high mortality. In addition, EBV infection is also associated with chronic hepatitis, liver cirrhosis, autoimmune liver disease, etc.

[Emphasizing the clinical diagnosis and treatment of liver disease-associated infections].

Liver disease-associated infection is a common condition in clinic. The existence of underlying liver disease increases the morbidity, mortality, and other adverse consequences of various infections, which in turn aggravates the liver disease-associated infections progression. Therefore, liver disease and infection should be emphasized in the clinical management. The assessment of liver disease includes its etiology, severity level, and complications, while the assessment of infectious diseases emphasizes its etiology and lesion location. Timely elimination of pathogens and complications is the treatment aspect of liver disease, whereas empiric pathogenic therapy is the treatment aspect of infection. The use of anti-infective drugs with a high risk of liver damage should be avoided when the pathogen is determined, and the target should be treated as soon as possible.

[Retrospective analysis of risk factors of liver cirrhosis combined with overt hepatic encephalopathy: a single -center case-control study].

Objective: To explore the risk factors of overt hepatic encephalopathy (OHE) in patients with liver cirrhosis. Methods: A retrospective study was designed. Patients with liver cirrhosis combined with /without OHE who were hospitalized to our hospital during the same period were selected as the case/control group. Clinical and laboratory data of both groups of patients were compared to analyze the risk factors affecting the occurrence of OHE. SPSS software was used for statistical analysis. A t-test or rank-sum test was used to compare the measurement data. Chi-square test or Fisher's exact probability method was used to compare the count data. Logistic regression was used for multivariate analysis. Results: A total of 500 patients with liver cirrhosis diagnosed in our hospital from August 2017 to December 2018 were selected as the case group, and 40 cases with cirrhosis without OHE who were hospitalized during the same period were randomly selected as the control group. The gender composition and age of the case and control group were comparable. Viral hepatitis (mainly hepatitis B) was the main etiology of liver cirrhosis in both groups. There were 52.5% patients in the case group and 57.5% patients in the control group, respectively. Alcoholic liver disease, autoimmune liver disease and so on were the other included causes. With regard to blood biochemical indicators, the serum creatinine levels of both groups were comparable, but in the case group, the serum total bilirubin level was higher (34.30 µmol / L vs. 18.65 µ mol/L, Z = -3.185, P < 0.05), while the serum sodium level was lower (137.00 mmol/L vs. 140.08 mmol/L, Z = -2.348, P < 0.05), and the prothrombin time was longer (14.60 s vs. 12.20 s) s. 078, P < 0.05), and international normalized ratio (1.33 vs. 1.07, Z = - 5.632, P < 0.05), and serum albumin level (30.6 g/L vs. 35.6 g/L, t = 3.386, P < 0.05) was lower. In terms of complications, patients in the case group had a higher proportion of combined gastrointestinal bleeding (30.0% vs. 10.0%, χ(2) = 5.000, P < 0.05), ascites (87.5% vs. 30.0%, χ(2) = 27.286, P < 0.05) and secondary infection (32.5% vs. 10.0%, χ(2) = 7.813, P < 0.05). In terms of severity classification, the proportion of Child-Pugh C in the case group was higher (62.5% vs. 10.0%, χ(2) =26.593, P < 0.05). In terms of outcome, there were 3 deaths in the case group and no deaths in the control group. Multivariate analysis showed that Child-Pugh class C (OR = 12.696), and combined ascites (OR = 10.655) were an independent risk factor for OHE in patients with liver cirrhosis. Conclusion: Our single-center retrospective clinical study shows that patients with cirrhosis combined with OHE are more critical and have more complications. In order to promptly diagnose and treat OHE, more attention should be paid to patients with combined ascites and Child-Pugh class C.

[Effects of programmed death receptor-1 antibody in patients with hepatitis B-associated liver cancer].

Objective: To investigate the effect of programmed death receptor (PD)-1 antibody therapy in patients with hepatitis B-associated liver cancer. Methods: Data of 29 chronically infected HBV patients with liver cancer who received PD-1 antibody combined with tyrosine kinase inhibitor in the Department of Infectious Diseases of the Fifth Medical Center of PLA General Hospital from March 2020 to January 2021 were selected. At the same time, all of the above-mentioned hepatitis B virus (HBV) patients were treated with nucleos(t)ide analogues. Patients clinical diagnostic data, laboratory test results, tumor response and the incidence of adverse reactions were collected retrospectively to understand the overall safety, therapeutic anti-tumor effect, HBV changes condition and the correlation between HBV changes and anti-tumor PD-1 antibody efficacy, high viral load treatment condition, and HBV reactivation safety issues. Statistical analysis was performed by non-parametric rank sum test. Results: Therapeutic anti-tumor effect and safety profile were good in patients. The complete remission rate was reached 27.6%. Adverse reactions were mostly mild, and the incidence of serious adverse reactions was low. After 12 weeks of follow-up, HBV DNA and hepatitis B surface antigen (HBsAg) was quantitatively decreased (P < 0.05). HBV DNA and HBsAg were decreased more significantly in patients with progressive disease (PD), stable disease (SD) and partial response (PR) (P < 0.05). Five patients with HBV DNA ≥ 10(4) IU/ml had responded well to the tumor treatment without serious adverse reactions. One patient had a slight increase in HBV DNA and alanine aminotransferase, while there was no HBV reactivation and correlated liver damage. Conclusion: Patients with HBV-associated liver cancer who received combined therapy have good anti-tumor efficacy and safety profile. PD-1 treatment has a certain effect on HBV. Compared with non-responders, patients with tumor response have better antiviral treatment efficacy. The safety of treatment in patients with high viral load is manageable, and there are no safety issues related to HBV reactivation.

[High frequency hearing loss detection rate in occupational noise exposed workers in China: a Meta-analysis].

Objective: A Meta-analysis was used to evaluate the high frequency hearing loss detection rate, its area and time distributions in occupational noise exposed workers in China to provide evidence for the prevention and control of occupational noise-induced deafness. Methods: From February to April 2020, the number of hearing tests and the number of binaural high frequency average hearing threshold ≥40 dB of workers exposed to noise in key occupational disease monitoring and occupational health risk assessment project in China was collected from China National Knowledge Infrastructure (CNKI) , VIP database and PubMed database. The retrieval time was set from the establishment of the database to February 28, 2020. The R 3.6.2 software was used to calculate the pooled high frequency hearing loss detection rate. Results: A total of 28 studies, included 34 data and 1259571 occupational noise exposed workers, were enrolled. The articles covered 16 provinces in 2015-2018. The pooled high frequency hearing loss detection rate in occupational noise exposed workers was 9.45% (95%CI: 7.64%-11.64%) . The high frequency hearing loss detection rate (10.54%) was higher in 2018 than other years (2017: 10.17%, 2016: 9.21%, 2015: 7.88%) . The high frequency hearing loss detection rate (12.23%) was higher in central China than in eastern (10.21%) , western (7.91%) and north-eastern (5.21%) China. Conclusion: The overall high frequency hearing loss detection rate in occupational noise exposed workers is generally high in China. It is important to strengthen the hearing protection in occupational noise exposed workers to facilitate the occupational noise-induced deafness prevention and control in China.

[Knockdown of MTA1 inhibits the proliferation and apoptosis of esophageal cancer cells].

Objective: To investigate the effects of metastasis associated gene 1 (MTA1) expression on the proliferation and apoptosis of human esophageal cancer Eca109 cells. Methods: MTA1 siRNA was transfected into human esophageal cancer Eca109 cells, and the control group and blank group were set up. The expression of MTA1 in Eca109 cells with different treatment was detected by real-time fluorescent quantitative PCR (RT-PCR) and western blot. The proliferation of Eca109 cells was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. The cloning formation ability of Eca109 cells was detected by plate cloning assay. The apoptosis of Eca109 cells were detected by flow cytometry. The expression of proliferating cell nuclear antigen (PCNA) and apoptosis-related proteins, including cleaved caspase-3 and total caspase-3 protein in Eca109 cells were detected by western blot. Results: After 48 hours of transfection, RT-PCR result showed that the relative expression levels of MTA1 mRNA in Eca109 cells in the blank group, control group, and siRNA group were 1.00±0.10, 0.98±0.09 and 0.21±0.03, respectively. The expression of MTA1 mRNA in siRNA group was significantly inhibited (P<0.05), while no significant difference of MTA1 mRNA expression between the blank group and the control group has been found (P>0.05). Western blot results were consistent with those of RT-PCR. MTT array results showed that, compared with the blank group and transfection group, the absorbance values of Eca109 cells in siRNA group were dramatically reduced at 48, 72, and 96 h (all P<0.05). There were no significant differences of absorbance values between the blank group and the control group at 48, 72, and 96 h (all P>0.05). The results of the plate colony formation test showed that the number of colony formation in the blank group and control group were 58.64±6.86 and 60.02±7.04, respectively, significantly higher than 18.10±3.16 in siRNA group (P<0.05), while there was no significant difference between the blank group and control group (P>0.05). Flow cytometry results showed that the apoptosis rates in the blank group and control group were (2.13±0.54)% and (2.27±0.61)%, respectively, significantly lower than (32.61±5.28)% in siRNA group (P<0.05), while there was no significant difference between blank group and control group (P>0.05). Western blot results showed that the expression of PCNA protein was down-regulated while cleaved caspase-3 protein expression was upregulated in siRNA group, compared to the control group and blank group (P<0.05). Conclusions: Inhibition of MTA1 expression can inhibit the proliferation of Eca109 cells and induce apoptosis. This process may be related to the down-regulation of PCNA protein expression and activation of caspase-3 protein expression.

Donor-derived marrow mesenchymal stromal cell co-transplantation following a haploidentical hematopoietic stem cell transplantation trail to treat severe aplastic anemia in children.

Haploidentical hematopoietic stem cell transplantation (haplo-HSCT) is associated with an increased risk of graft failure and severe graft-versus-host disease (GVHD). Recent studies have shown that mesenchymal stromal cells (MSCs) display potent immunosuppressive effects and can support normal hematopoiesis. In a multi-center trial, we co-transplanted culture-expanded donor-derived bone marrow MSCs (BM-MSCs) into 35 children with severe aplastic anemia (SAA) undergoing haplo-HSCT. All 35 patients (100%) achieved hematopoietic reconstitution and showed sustained full donor chimerism. The median time for myeloid engraftment was 14 days (range 10-22 days), while that for platelet engraftment was 18 days (range 9-36 days). The incidence of grade II-IV acute GVHD and chronic GVHD was 25.71 and 22.86%, respectively. The overall survival rate was 85.71% with a median of 22 months (range 3.5-37 months). The combined transplantation of haploidentical HSCs and BM-MSCs into children with SAA without an HLA-identical sibling donor is relatively safe and may represent an effective new therapy to improve survival rates and reduce the risk of graft failure.

Visual and rapid detection of Plesiomonas shigelloides using loop-mediated isothermal amplification method.

Plesiomonas shigelloides is a common pathogen of aquatic animals and can pose a certain hazard to aquaculture. Here, we aimed to develop a loop-mediated isothermal amplification (LAMP) method for the visual detection of P. shigelloides to aid the diagnosis of infections caused by this pathogen in aquatic animals. We used LAMP to amplify P. shigelloides DNA and combined it with calcein or nucleic acid dipstick assay (NADA) to visualize the amplified products. The optimal LAMP amplification temperature was 64°C, and the reaction lasted for 50 min. The limit of detection of recombinant plasmids containing the target gene using the LAMP method was 2·0 × 102 copies per µl, which is ten times higher than that using conventional polymerase chain reaction (PCR). LAMP products could be visualized without agarose gel electrophoresis. We tested 85 fish specimens using the established LAMP method and conventional PCR. The detection rate was 42·4% using the LAMP method and 34·1% using conventional PCR. Based on our results, the LAMP method combined with calcein or NADA is a rapid, specific, sensitive and accurate method for visual detection of fish-derived P. shigelloides and can be used for the laboratory diagnosis of infections caused by it. SIGNIFICANCE AND IMPACT OF THE STUDY: The combination of loop-mediated isothermal amplification (LAMP) and calcein and nucleic acid dipstick assay (NADA) provided a rapid, specific and sensitive method for detecting Plesiomonas shigelloides, which is an important pathogen that causes diseases in aquatic animals worldwide. In the present study, the LAMP method showed a higher detection rate than conventional PCR for P. shigelloides using templates from 85 fish specimens. Thus, the LAMP method could be a reliable and convenient tool for diagnosing diseases in aquatic animals in the laboratory.