OBJECTIVE: To explore the mutation of Y-STR loci in meiotic allelic transmission in a large pedigree. METHODS: The oral swabs of 163 male individuals were collected from a Lin pedigree. Twenty-two Y-STR genetic markers were typed with AGCU Y24 fluorescent detection kit (AGCU Y24 system), which also contained 16 Y-STR markers included in Yfiler multiple amplification kit (Yfiler system). The genotyping results of Y-STR loci were compared between each two males in the pedigree. RESULTS: There were 20 and 30 kinds of haplotypes obtained with Yfiler and AGCU Y24 systems in 163 male individuals from the Lin pedigree, respectively. The rates referred to haplotype differences (RRHD) of these two typing systems between male pairs were 0.910 5 and 0.922 7, respectively. The average number of marker differences were 6.582 1 and 9.824 8, respectively. The RRHD increased along with the incidents of meiosis. CONCLUSION Y-STR mutation leads to different Y-STR haplotypes among the male members in a paternal pedigree and the rate of difference increases along with the incidents of meiosis.
Alleles ; Chromosomes, Human, Y/genetics* ; DNA Fingerprinting ; Genetic Linkage ; Genetic Markers/physiology* ; Genotype ; Haplotypes ; Humans ; Male ; Mutation/genetics* ; Pedigree

Objective To evaluate the efficacy of the combination of percutaneous transhepatic embolization of gastroesophageal varices (PEGV) and partial splenic embolization(PSE) for the treatment of variceal bleeding. Methods Fifty patients with cirrhosis who fit in with the requirements of the study were randomly divided into dual-interventional group and surgical group. The patients in dual-interventional group were treated with PEGV together with PSE, and the patients in surgery group were treated with Hassab's operation. After the procedure all the patients in two groups were followed up periodically. The endoscopy, B ultrasonography, liver function tests and hematologic examinations were performed 24 months after the therapy, and the results were statistically analyzed. Results Fifty patients were enrolled in this study. The procedures of embolization and surgery were successful in all patients. In dual-interventional group, the whit eblood cell and platelet counts were (2.33±0.65) 10~9/L and (3.63±1.05) ×10~9/L respectively before the treatment and were (7.98±3.0) ×10~9/L and (163±91)× 10~9/L respectively 24 months after the treatment (P<0.05). The diameter of theportal vein was (1.47±0.25) cm before the treatment and was(1.31±0.23) cm 24 months after the treatment (P<0.05). The severity of esophageal varices decreased from grade Ⅲ to lower grade Ⅱ in 11 patients, and from grade Ⅱ to lower grade Ⅰ in 6 patients 24 months after procedure. Portal thrombosis occurred in 1 case. The recurrent bleeding rate was 16% (4/25) 24 months after treatment. Three patients died of the recurrent bleeding, one patient died of hepatic failure. In surgical group, the white blood cell and platelet counts were (2.2±0.60) ×10~9/L and (41±12.5) ×109/L before treatment, and were (9.3±2.56)×10~9/L and (321±12.5)×10~9/L 24 months after treatment (P<0.05). The diameter of the portal vein was (1.43±0.22) cm before the treatment and was (1.28±0.18) cm after the treatment (P<0.05). The severity of esophageal varices decreased from grade Ⅲ to lower grade Ⅱ in 13 patients, and from grade Ⅱ to lower grade Ⅰ in 7 patients. Four patients developed portal thrombosis after the procedure. The recurrent bleeding rate was 20%(5/25), 2 patients died of hepatic failure after the surgery, 2 died of recurrent bleeding. Conclusion In treating patients with cirrhosis, the combination of PGEV and PSE, regarded as dual-interventional therapy, is very effective, especially in controlling recurrent bleeding and in improving white blood cell and platelet counts.

Objective:To explore the clinical features of decompensated liver cirrhosis patients with acute kidney injury (AKI) progressing to chronic kidney disease (CKD) and its impact on prognosis.Methods:From January 2015 to July 2019, at Tianjin Second People′s Hospital, the general data and laboratory test results of 346 hospitalized patients with decompensated liver cirrhosis were retrospectively analyzed. The patients were followed up for 12 months. Univariate analysis and multivariate logistic regression analysis were used to analyze the risk factors of AKI and CKD. Kaplan-Meier method was used for survival analysis. The independent sample t test, Mann-Whitney U test and chi-square test were used for statistical analysis. Results:A total of 128 patients with decompensated liver cirrhosis developed AKI, and 25 of them developed into CKD. Univariate analysis showed that age, hypertension, complications of liver cirrhosis infection, ascites and hepatic encephalopathy, acute-on-chronic liver failure, Child-Turcotte-Pugh score of liver function, baseline serum creatinine, post-admission serum creatinine, serum sodium, white blood cell count, total cholesterol, triglyceride, high-density lipoprotein, total bilirubin, albumin, international normalized ratio (INR) and prothrombin time activity were risk factors of AKI in patients with decompensated liver cirrhosis ( t=3.822, χ2=12.534, 26.761, 5.035, 3.894 and 26.101, U=7 004.500, 9 132.500, 5 925.000, 10 144.000, 10 717.500, 10 827.000, 10 912.000, 5 741.500, 10 017.000, 10 187.500, 11 680.500 and 11 321.500, all P<0.05). The risk factors of AKI progressing to CKD in decompensated liver cirrhosis included the etiology of liver cirrhosis, hypertension, baseline serum creatinine, serum creatinine at the time of diagnosis of AKI, total cholesterol, INR, AKI etiology and AKI classification ( χ2=13.153 and 9.144, U=353.000, 337.000, 576.500 and 481.000, χ2=9.501 and 17.801, all P<0.05). The results of multivariate logistic regression analysis showed that the independent risk factors of AKI progressing to CKD in decompensated liver cirrhosis included baseline serum creatinine (odds ratio ( OR)=1.066, 95% confidence interval ( CI) 1.020 to 1.114, P=0.005) and AKI classification ( OR=6.086, 95% CI 1.828 to 20.260, P=0.003). The Kaplan-Meier survival curve showing that after following up for 12 months, the survival rate of patients with decompensated liver cirrhosis patients who progressed to CKD from AKI was lower than that of patients who did not developed into CKD (52.0%, 13/25 vs. 86.4%, 51/59), and the difference was statistically significant ( χ2=11.482, P=0.001). Conclusion:The transition from AKI into CKD is common in patients with liver cirrhosis, which affects the clinical prognosis and reduces the survival rate.

Objective:To analyze the clinical characteristics and causes of death of 80 dead cases with confirmed coronavirus disease 2019 (COVID-19).Methods:The clinical data of 80 dead patients with COVID-19 who were admitted to Renmin Hospital of Wuhan University from January 11 to February 11, 2020 were retrospectively analyzed.The laboratory examination indexes (including white blood cells, lymphocytes, procalcitonin (PCT), lactic acid, D-dimmer, fibrinogen degradation products, N-terminal pro-brain natriuretic peptide (N-proBNP), ultra sensitive-troponin I, lactate dehydrogenase (LDH) and CD4 + T lymphocyte) of the patients at the time of admission were compared with the indexes at the last time before death. Statistical analysis was conducted by using paired t test or Wilcoxon′s signed rank test. Results:The median age was 72 years old of the 80 patients, and 78.75%(63/80) of them were older than 60 years. Thirty-six cases (45.00%) were severe and 44(55.00%) were critical at admission. Fifty-eight cases (72.50%) had underlying diseases. The common underlying diseases were hypertension, diabetes mellitus, coronary atherosclerotic heart disease, and chronic obstructive pulmonary disease. Comparing the patients′ first laboratory tests at admission with those before death, white blood cells increased (8.01(4.86, 12.29)×10 9/L vs 12.55(8.25, 17.66)×10 9/L), lymphocytes decreased (0.70(0.46, 0.88)×10 9/L vs 0.54(0.39, 0.75)×10 9/L), PCT increased (0.20(0.11, 0.74) μg/L vs 1.00(0.20, 1.99) μg/L), lactic acid increased (2.10(1.40, 3.10) mmol/L vs 3.10(2.60, 4.10) mmol/L), D-dimmer increased (4.33(0.97, 18.98) mg/L vs 15.29(5.17, 53.44) mg/L), fibrinogen degradation products increased (15.90(3.58, 76.60) mg/L vs 63.14(21.23, 110.67) mg/L), N-proBNP increased (1 078.00(347.35, 2 996.50) ng/L vs 3 439.50(1 576.00, 9 281.50) ng/L), ultra-sensitive troponin I increased (0.08(0.03, 0.17) μg/L vs 0.33(0.14, 2.47) μg/L), LDH increased (397.00(327.00, 523.50) U/L vs 624.00(481.00, 854.00) U/L) and CD4 + T lymphocyte decreased (137.00(104.00, 168.00)/μL vs 97.00(67.00, 128.00)/μL). The differences between the two groups were all statistically significant ( W=238.00, 1 053.50, 150.00, 152.00, 192.00, 190.00, 108.00, 57.00, 53.00 and 40.00, respectively, all P<0.05). All patients received antiviral and respiratory-support therapy and the main cause of death was respiratory failure caused by intractable hypoxemia and multiple organ failure. Among them, seven cases died in one day hospitalization, and 66 cases died in seven days hospitalization. Conclusions:Elderly patients with a variety of chronic underlying diseases have poor prognosis. It′s essential to pay more attention and deal with the above clinical characteristics at an early stage to improve the outcome of the COVID-19 patients.

OBJECTIVE: To explore the molecular mechanisms of colonic epithelial aging related proteins and aged colonic epithelial susceptibility to tumor. METHODS: The proteins of normal human colonic epithelial tissue from young and old people were separated by 2-dimensional gel electrophoresis (2DGE), respectively. Then gels were stained by silver, scanned by imagescanner and analyzed with PDQuest software. The differentially expressed protein spots of colonic epithelium between the old and the young groups were identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. RESULTS: Well-resolved and reproducible 2DGE maps of normal human colonic epithelium from the young and the old were acquired. Nineteen more than 2 fold differentially expressed protein spots were identified representing 17 different proteins by MALDI-TOF-MS. The functions of these proteins involve in metabolism, energy generation, transportation, antioxidation, translation and protein folding. CONCLUSION Seventeen aging related proteins of human colonic epithelium identified indicate that injury of mitochondrial function and decline of antioxidant capability are important reasons for the aging of human colonic epithelium. These data provided useful clues for elucidating the mechanisms of colonic epithelial aging and aged colonic epithelial susceptibility to cancer.
Aging ; metabolism ; Cells, Cultured ; Cellular Senescence ; genetics ; Chloride Channels ; biosynthesis ; genetics ; Colon ; cytology ; Electron-Transferring Flavoproteins ; biosynthesis ; genetics ; Epithelial Cells ; cytology ; Humans ; Intestinal Mucosa ; cytology ; Proteins ; metabolism

OBJECTIVE: To investigate the proteome of hepatocyte transformation by hepatitis C virus (HCV) nonstructural protein 3 (NS3). METHODS: Human hepatocyte line QSG7701 stably expressing HCV NS3 C-terminal deleted protein was constructed, which was named pRcHCNS3/QSG. Two-dimensional electrophoresis (2-DE) was used to separate the total protein of pRcHCNS3/QSG and pRcCMV transfected cells (pRcCMV/QSG) respectively. Differentially expressed protein spots were identified by mass spectrometry. Western blot confirmed the differentially expressed proteins. RESULTS: 2-DE profiles with high resolution and reproducibility were obtained. The average spots of pRcHCNS3/QSG and pRcCMV/QSG were (1183+/-77) and (1095+/-82) respectively, and (920+/-60) spots were matched. Twenty-one differentially expressed protein spots were chosen randomly and 15 were identified by mass spectrometry. Some proteins such as Ras, P38 and HD53 which were involved in signal transduction were increased in pRcHCNS3/QSG cells. Western blot also showed strong expression of phosphorylated P44/42 and P38 in pRcHCNS3/QSG cells. Other differentially expressed proteins were related to cell cycle regulation, immunoreaction, tumor invasion and metastasis, and liver metabolizability. CONCLUSION HCV NS3 might be involved in cell malignant transformation through affecting protein expression and signal transduction such as MAPK cascade. Further study on the signal transductions and their relationship would not only be helpful to explore the mechanism of HCV related HCC, but also provide a new idea for the molecular treatment of HCC.
Cell Line ; Cell Transformation, Neoplastic ; Electrophoresis, Gel, Two-Dimensional ; methods ; Hepatocytes ; metabolism ; pathology ; Humans ; Mass Spectrometry ; methods ; Proteome ; analysis ; Proteomics ; methods ; Transfection ; Viral Nonstructural Proteins ; biosynthesis ; genetics

Objective:To investigate the effects of different chemotherapy dose intensity on the short-term efficacy and adverse reactions of patients with advanced colon cancer.Methods:A real-world database of patients with advanced colon cancer in Wangjing Hospital of China Academy of Chinese Medical Sciences and China-Japan Friendship Hospital from January 2017 to December 2020 was established, including 105 patients treated with the same chemotherapy regimen for two consecutive cycles. The patients were grouped according to the average relative dose intensity (ARDI) of chemotherapy, and the population differences, treatment regimens, short-term efficacy and adverse reactions of different chemotherapy dose intensities were evaluated. The receiver operating characteristic (ROC) curve was used to analyze the predictive value of ARDI for short-term efficacy.Results:There were 31 patients in the high dose intensity group (ARDI≥80%) , 34 patients in the medium dose intensity group (80%0.05) . ROC curve analysis found that the area under the curve predicting objective response was 0.70, the sensitivity was 92.30%, and the specificity was 52.10% when the chemotherapy ARDI threshold was 64%. Conclusion:In the real world, colon cancer patients receiving high dose intensive chemotherapy have better objective response rate, and tumor markers decreased significantly. The baseline status of colon cancer patients receiving low dose intensive chemotherapy is poor, but there is no statistically significant difference in disease control rate between patients receiving low dose intensive chemotherapy and patients receiving high dose intensive chemotherapy, and the adverse reactions are controllable.

Objective To explore the predictive value of immune inflammation combined with liver function hematological indicators for the metastasis of colorectal cancer.Methods A retrospective analysis of clinical data of 133 patients with colorectal cancer was conducted.The patients were divided into three groups based on disease progression after 24 months of postoperative follow-up:non-metastasis group(n=38),liver metastasis group(n=43),and non-liver distant metastasis group(n=52).The immune inflammatory markers and liver function hematological indicators of progression-free survival were analyzed.Nomogram prediction models were constructed using univariate and multivariate logistic regression analyses to identify risk factors for metastasis of colorectal cancer.The accuracy of the nomogram was validated using receiver operating characteristic(ROC)curve and calibration curve,and the clinical predictive efficacy was evaluated through decision curve and clinical impact curve.Results Univariate and multivariate logistic regression analyses showed that pan-immune-inflammatory value(PIV),prognostic nutritional index(PNI),and bile acid(BA)were independent predictors of colorectal cancer metastasis.The area under the ROC curve of the combined prediction of metastasis was 0.84;neutrophil/lymphocyte ratio(NLR)and BA were independent predictors of liver metastasis from colorectal cancer.The area under the ROC curve of the combined prediction of liver metastasis was 0.83;PIV and PNI were independent predictive factors for the occurrence of non-liver distant metastasis from colorectal cancer.The area under the ROC curve for the combined prediction of non-liver distant metastasis was 0.83.The calibration curve,decision curve,and clinical impact curve showed that the three models had good accuracy and net benefit value.Conclusion The nomogram constructed based on immune inflammation and liver function hematological indicators can predict the metastasis of patients with colorectal cancer and has high predictive efficacy and clinical application prospects.

Human parathyroid hormone (hPTH) was highly expressed in Escherichia coli by inserted the synthesized whole hPTH cDNA into the vectors pBV220 and pET22b. After expression and disruption, the purified product was acquired through cation exchange chromatography and reverse phase chromatography. From the results of N-terminal sequencing and MALDI-TOF-MS analysis the recombiant prtein was indentified as intact hPTH. In in vitro Bioassays the recombinant hPTH stimulated adenylate cyclase as the standard did. In ovariectomized rats the recombinant hPTH markedly increased the femoral bone mass and bone mineral density.
Amino Acid Sequence ; Animals ; Base Sequence ; Bone Density ; drug effects ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Molecular Sequence Data ; Ovariectomy ; Parathyroid Hormone ; chemistry ; genetics ; metabolism ; pharmacology ; Rats ; Rats, Wistar ; Sequence Alignment ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo study the cultural method and identification of human umbilical vein endothelial cells (HUVECs), and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains(Tie-2) in HUVECs.

METHODSHUVECs were isolated from umbilical veins by the technique of irrigative digestion, and were cultivated in plates. The cells were identified by VIII monoclonal antibody. Tie-2 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and SABC immunocytochemistry.

RESULTSHUVECs could adhere to the plates completely after 24 hours, and confluence a monolayer 4-5 days later. The band of Tie-2 mRNA was obviously and the expression of Tie-2 protein was strongly positive by immunocytochemistry in HUVECs. The positive rate was over 85%.

CONCLUSIONHighly purified endothelial cells were isolated. And there were overexpression of Tie-2 in HUVECs.