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 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 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.

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.

Cells, Cultured ; EGF Family of Proteins ; Endothelial Cells ; Human Umbilical Vein Endothelial Cells ; Humans ; Immunoglobulins ; TYK2 Kinase ; Umbilical Veins

OBJECTIVETo prepare ITP plasma IgG and its F(ab')2 fragments and investigate their immunoreactivity to platelet GPIIb/IIIa and/or GPIb/IX and their effects on platelet aggregation function.

METHODSThe ITP patients having inhibitory autoantibody to the platelet aggregation were selected by modified MAIPA and platelet aggregation test with turbidimetry. Plasma IgG and its F(ab')2 fragments were prepared by streptococcal protein A affinity column and pepsin digestion. The immunoreactivity and the effects on platelet aggregation function of the whole antibody and its fragments were detected by modified MAIPA and platelet aggregation test, respectively.

RESULTS(1) Anti-platelet GPIIb/IIIa and/or GPIb/IX autoantibodies were detected in 34 of 68 (53.6%) ITP patients' plasmas and that from 5 patients significantly inhibited the platelet aggregation induced by ADP or ristocetin. (2) By using protein A column combined with protease digestion, pure IgG and its F(ab')2 fragments were successfully obtained. (3) The purified IgG and its F(ab')2 fragments retained the ability to bind to their respective glycoproteins and inhibited the platelet aggregation function, whereas the IgG depleted plasma lost the ability of binding to the platelet GPs.

CONCLUSIONSF(ab')2 fragment of the IgG antibody is a functional fragment, which not only has the binding ability to the platelet GPs but also inhibits the platelet aggregation function in a dose-dependent manner.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Humans ; Immunoglobulin Fab Fragments ; immunology ; Immunoglobulin G ; immunology ; Integrin beta3 ; immunology ; Male ; Middle Aged ; Platelet Aggregation ; Platelet Membrane Glycoprotein IIb ; immunology ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; physiopathology ; Young Adult

The roots, barks, branches and pericarps of Juglans mandshurica were used as folk medicine in China and reputed for its treatment of several cancers, such as gastric cancer, liver cancer and leukemia. The extracts of the roots, branches, leaves and pericarps of J. mandshurica have been experimentally proved to show anti-tumor activities. Tannins, which exhibited antioxidant and anti-tumor activities, were the main constituents in J. mandshurica. In this paper, a simple spectrophotometric method was developed for the determination of total tannins in the roots, branches, leaves and pericarps of J. mandshurica collected in Dalian and Anshan of Liaoning Province. Gallic acid was used as standard compound and the content of total tannins was calculated as gallic acid equivalent. As a result of the method validation, a good linearity (r = 0.9997, n = 5) and a high recovery of gallic acid (99.02%, RSD 3.7%, n = 9) was achieved. Eight samples including four parts of J. mandshurica collected in two places were analyzed for their total tannins with the established method. In the corresponding parts of J. mandshurica, except the pericarps, the contents of total tannins showed no significant difference between samples collected in Dalian and Anshan, while the content of total tannins in different parts of J. mandshurica were significantly different. The average content of total tannins in the roots, branches, leaves and pericarps of samples collected in Dalian and Anshan was 45.66, 23.40, 58.24, 3.58 mg g(-1), respectively.
Juglans ; chemistry ; Plant Extracts ; analysis ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Tannins ; analysis

OBJECTIVETo study the anti-platelet GPVI single chain Fv phage antibody which can inhibit the aggregation function of platelet by using phage antibody library technology.

METHODSITP patients with anti-platelet GPVI autoantibody that could inhibit the aggregation function of platelet were screened by MAIPA assay and platelet aggregation test. The gene fragments of heavy chain and light chain variable region (VH and VL) of immunoglobulin were amplified by RT-PCR from peripheral blood lymphocytes mRNA of the screened patients. The VH and and VL fragments were linked through a DNA linker encoding the peptide (Gly4Ser)3 to construct single chain Fv (ScFv) gene. The ScFv gene was digested with SfiI/NotI restriction enzymes and cloned into the pHEN2 phage display vector, then electrically transformed to E. coli TG1. The TG1 containing ScFv-pHEN2 was rescued by helper phage M13K07 to produce ScFv phage antibody. The anti-platelet GPVI phage ScFv antibody was enriched and purified. The effect of the phage antibody on platelet aggregation function was studied.

RESULTSOf 806 chronic ITP patients, 11 (1.36%) were positive for anti-platelet GPVI autoantibody and 2 (0.24%) patients'plasma significantly inhibited the collagen induced platelet aggregation. The length of VH and VL fragments was about 380 to 400 bp, and were successfully formed ScFv fragments of about 800 bp by DNA linker. After cloning ScFv to phagemid vector pHEN2 and transforming ScFv-pHEN2 to TG1, 4.1x10(7) clones were obtained. After M13K07 rescue, 2.62x10(10) cfu/ml ScFv phage antibodies were produced. The purified anti-platelet GPVI ScFv phage antibody inhibited the collagen induced platelet aggregation.

CONCLUSIONAnti-platelet GPVI ScFv phage antibody produced by phage antibody library technology can inhibit the aggregation function of platelet.

Adolescent ; Adult ; Aged ; Escherichia coli ; genetics ; Female ; Humans ; Male ; Middle Aged ; Peptide Library ; Platelet Aggregation ; immunology ; Platelet Aggregation Inhibitors ; immunology ; pharmacology ; Platelet Membrane Glycoproteins ; immunology ; Single-Chain Antibodies ; genetics ; immunology ; pharmacology ; Transformation, Bacterial ; Young Adult

OBJECTIVETo explore the regulative efficacy of Pu'er tea () extract on metabolic syndrome.

METHODSNinety patients with metabolic syndrome were randomly divided into two groups, the intervention group administered with Pu'er tea extract, and the placebo group with placebo capsules. After 3 months' treatment, body mass index, waist hip ratio, blood lipids, blood sugar, immune and inflammatory index, and oxidation index of the patients with metabolic syndrome were tested and analyzed.

RESULTSIn the intervention group, the body mass index, waist-hip ratio, fasting and 2 h postprandial blood glucose, serum total cholesterol, triglycerides, low density lipoprotein and apolipoprotein B-100 all decreased in the patients with metabolic syndrome, and also the high-density lipoprotein level increased and apolipoprotein A-1 showed the tendency to increase. Serum C-reactive protein, tumor necrosis factor-α, and interleukin-6 were decreased in the intervention group. Interleukin-10 level was increased, MDA was decreased and superoxide dismutase was increased. Compared with before treatment and the placebo group, there were significant differences (P<0.05, P<0.01).

CONCLUSIONSPu'er tea demonstrated excellent potential in improving central obesity, adjusting blood lipid, lowering blood sugar, regulating immunity and resisting oxidation. It can adjust the metabolic syndrome of different clinical phenotypes to different degrees, and is ideally fit for early prevention of metabolic syndrome.

Adult ; Aged ; Blood Glucose ; metabolism ; Body Mass Index ; Double-Blind Method ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Humans ; Inflammation ; blood ; complications ; Lipids ; blood ; Male ; Malondialdehyde ; blood ; Metabolic Syndrome ; blood ; complications ; drug therapy ; Middle Aged ; Oxidation-Reduction ; Placebos ; Plant Extracts ; therapeutic use ; Superoxide Dismutase ; blood ; Waist-Hip Ratio