OBJECTIVE: To extract sperm DNA from mixed stain by the modified differential lysis method combined with silicon bead method and to evaluate its application value. METHODS: Fifty-two mixed stains containing female STR genotypes detected by differential lysis method were collected. The sperm DNA was extracted by the modified method combined with silicon bead method, then genotyped with the Identifiler Kit, and compared with the results of genotyping by the conventional differential lysis method as control. RESULTS: Of the 52 samples, 38 samples with sole male STR genotypes in all loci were detected. The detection rate of male STR genotypes was 98.08% through the modified method combined with silicon bead method. CONCLUSION The modified differential lysis method combined with silicon bead method can be used in extraction of sperm DNA from mixed stain.
DNA/isolation & purification* ; DNA Fingerprinting ; Female ; Genotype ; Humans ; Male ; Polymerase Chain Reaction ; Silicon ; Spermatozoa

Adult ; Aged ; Aged, 80 and over ; Epistaxis ; therapy ; Female ; Gloves, Protective ; Hemostatic Techniques ; Humans ; Male ; Middle Aged

Objective To establish a method of detecting hepatitis B virus x antigen (HBxAg) and antibody to HBxAg (anti-HBx) and to demonstrate its clinical significance of HBxAg and anti-HBx in sera from patients of chronic hepatitis B (CHB),liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Methods Full length HBx gene was cloned into pET30a(+),a prokaryotic expression vector,named pET30a-X.It was transformed into Escherichia coli BL21 (DE3),followed the fusion protein of HBx-His was induced by IPTG.The purified fusion protein was used to immunize rabbit as an antigen to generate polyclonal antibody to HBx protein.The method of enzyme-linked immunosorbent assay (ELISA) was established by using purified fusion protein and generated antibody,which was used to detect HBxAg and anti-HBx in sera from patients of CHB,LC,HCC and normal healthy people.Results The positive rates of HBxAg/anti-HBx were 8.7%/10.4% for CHB,17.9%/40.6% for LC,and 9.8%/34.4% for HCC, respectively.In statistics,the positive rates of anti-HBx in LC and HCC were higher than that in CHB (P

BACKGROUND: Vertebroplasty (PVP) and kyphoplasty (PKP) are important methods for the treatment of osteoporotic vertebral compression fractures in the elderly. Although bone cement has certain liquidity and vertebral fractures are often in different situations, bone cement leakage rate is still high. OBJECTIVE: Based on the theoretical discussion and clinical analysis, to study the theoretical causes of bone cement leakage and effective prevention methods. METHODS: A total of 162 cases (186 vertebrae) were treated with three methods of vertebroplasty. Group A: 64 cases with 78 vertebrae were treated with conventional cemented vertebroplasty; Group B: 57 cases with 65 vertebrae were treated with cemented vertebroplasty using cement pump; Group C: 41 cases with 43 vertebrae were treated with balloon kyphoplasty. The leakage of bone cement was observed in three groups. RESULTS AND CONCLUSION: Of the 186 vertebrae, postoperative bone cement leakage occurred in 25 vertebrae, with the leakage rate of 13% (25/186). Group A had bone cement leakage in 11 vertebrae, and the leakage rate was 14% (11/78). Group B had bone cement leakage in 8 vertebrae, and the leakage rate was 12% (8/65). Group C had bone cement leakage in 6 vertebrae, and the leakage rate was 14% (6/43). There was no significant difference among the three groups in the leakage rate of bone cement (P > 0.05). That is to say, the causes of bone cement leakage are not completely controllable, and the leakage position has some randomness. Strict and careful imaging monitoring is an intuitive method to prevent bone cement leakage.

OBJECTIVETo study the influence of separation of distal tibiofibular syndesmosis on ankle joint and to compare various operative methods so as to find suitable stabilization for separated distal tibiofibular syndesmosis.

METHODSFrom July 1997 to July 2002, we treated 87 patients (64 males and 23 females, aged 18-54 years) with separation of distal tibiofibular syndesmosis, among whom, 79 were combined with fracture of malleolus. Manipulative reduction, internal fixation with cancellous screws and external fixation with plaster support were performed on 37 patients, fixation with plate and screws for fibular fracture and fixation with cancellous screws for distal tibiofibular syndesmosis on 34 patients, and repair of the distal tibiofibular ligaments with tendon of peroneus longus, reduction of the separated distal tibiofibular syndesmosis, and fixation with cancellous screws on 16 patients. The ankle joint had been dorsiflexed for 30 degrees when the distal tibiofibular syndesmosis was fixed with cancellous screws. And the cancellous screws were taken out at 8-10 weeks after operation.

RESULTSThese patients were followed up for at least two years. The curative effects were assessed according to the complaints of the patients and the contour, function and radiogram of the ankle joint: excellent in 55 patients (63%), good in 18 patients (21%), and fair in 14 patients (16%). Separation of distal tibiofibular syndesmosis recurred in 2 patients, who underwent a reoperation for repairing the distal tibiofibular ligaments with tendon of peroneus longus and recovered. One cancellous screw was broken off. No necrosis developed in the anterior skin of the ankle mortise.

CONCLUSIONSSeparation of distal tibiofibular syndesmosis can be treated with various reasonable operations. Repair with tendon of the peroneus longus can get excellent outcomes for complete separation of the distal tibiofibular syndesmosis.

Adolescent ; Adult ; Ankle Injuries ; diagnostic imaging ; physiopathology ; surgery ; Bone Screws ; Casts, Surgical ; Female ; Fibula ; injuries ; physiopathology ; Follow-Up Studies ; Humans ; Ligaments, Articular ; injuries ; physiopathology ; Male ; Middle Aged ; Radiography ; Tibia ; injuries ; physiopathology ; Treatment Outcome

Objective:To investigate the effectiveness of surgical treatment for patients with T 4 stage prostate cancer. Methods:The clinical data and prognosis of 18 patients with T 4 stage prostate cancer treated in Shanghai Tenth People's Hospital from July 2013 to December 2019 were retrospectively analyzed. The average age of these 18 patients was 68.3 (53-81)years. 10 patients were castration resistant prostate cancer (CRPC) and 8 patients were hormone-sensitive prostate cancer (HSPC). 10 CRPC patients were treated with surgical treatment due to bladder clot packing and/or lower urinary tract obstruction. 8 HSPC patients had severe hematuria, severe dysuria and local symptoms. The KPS scores of all patients were ≥80 points with an average score of 84 (80-90). The average QOL score of 18 patients was 28 (21-32). 2 cases in 18 patients underwent total pelvic resection for rectal invasion (one CRPC and one HSPC). 7 cases underwent radical cystoprostatectomy for ureteral invasion (5 cases of CRPC, 2 cases of HSPC), 9 cases underwent bladder preservation surgery for bladder neck invasion (4 cases of CRPC, 5 cases of HSPC), of which 4 cases of enlarged lymph node dissection were all HSPC patients. Results:All cases of T 4 stage prostate cancer patients operation were successfully completed, the average operation time was 256 (219-310)min and the median intraoperative blood loss was about 300 (250-350)ml. Four of them (3 cases of CRPC and 1 case of HSPC) received blood transfusion after operation. The average postoperative hospital stay was 21(11-37) days. All 18 cases were followed up and the median follow-up time was 23.8 months. There was no perioperative death, and no bladder-preserving patients had true urinary incontinence or bladder outlet stenosis.2 CRPC cases died 8 and 15 months after surgery respectively, 7 patients were PSA relapse treated with docetaxel or abiterone therapy, and 1 HSPC patient with rectal invasion was followed up for 58 months after total pelvic resection, the PSA level was still 0.003ng/ml, no distant metastasis was found. 8 cases of hormone-sensitive patients were treated with endocrine therapy, and PSA was less than 0.2 ng/ml. The average QOL of 18 patients 3 months after operation was 37 points (25-45), which was significantly higher than that before operation. The average maximum urine flow rate of patients with bladder preservation was 23(19-25)ml/s. Conclusions:For T 4 stage prostate cancer, surgical treatment is feasible and safe for doctors with extensive surgical experience. For CRPC patients, the surgery can significantly improve short-term symptoms and quality of life, and long-term benefits need to be further evaluated with a large sample. For HSPC patients, it can not only improve clinical symptoms and QOL of patients, but also provide long-term benefits.

Objective To determine the predictive factors for antiviral therapy in hepatitis B e antigen (HBeAg)-negative chronic hepatitis B virus (HBV) infection [HBeAg(-) CHBI] patients with HBV DNA＜4.3 lg IU/mL.Methods A total of 179 HBeAg (-) CHBI patients were retrospective analyzed.Histology activity index (HAI) and fibrosis (S) were scored according to the Knodell HAI scoring system,and HAI＞3 and/or S≥3 was adopted as indications for treatment.Univariate and multiple regression analysis were used to assess factors associated with treatment indications.Receiver operating curves (ROC) and area under curve (AUC) were used to determine the predictive value of relevant factors.Results There were 81 cases with HAI＞3 (45.3％) and 72 with S≥3 (40.22％),and the proportion of patients with indications for treatment was 54.7 ％.Multiple regression analysis showed that age,γ-glutamyl transpeptadase (γ-GT),platelet (PLT) and albumin (Alb) were the predictive factors for the severity of liver damage and indication for treatment (all P＜0.05).The AUC for age,PLT,γ-GT and Alb were 0.655,0.657,0.726 and 0.65,respectively,and the corresponding Yoden index for age,PLT,γ-GT,and Alb were 0.297,0.426,0.03 and 0.012,respectively,the sensitivities of predicting HBeAg (-CHBI for treatment indications were 0.643,0.842,0.705 and 0.653,respectively.Conclusions This study shows that 54.7％ of HBeAg(-)CHBI patients with HBV DNA＜4.3 lg IU/ml have significant liver histological changes and require antiviral treatment.Older age,higher γ-GT,lower PLT and lower Alb levels are the predictive factors for treatment.

OBJECTIVETo analyze the correlation of Chinese medicine syndrome evolvement and cardiovascular: events in patients with stable coronary heart disease (CHD).

METHODSThis prospective cohort study investigated and: collected the clinical information of patients with stable CHD and observed the syndrome type at the baseline and 6-month at follow-up, as well as the cardiovascular events during the 6-month and 12-month follow-up. The patients were divided into the event group and the non-event group. The interaction and the impact of syndrome evolvement on cardiovascular events were examined through multifactor dimensionality reduction (MDR) analysis and the results were verified by Chi-square test.

RESULTSTotally 1,333 of 1,503 stable CHD patients enrolled met the inclusion criteria: of MDR analysis. Among them, 959 (71.9%) cases were males and 374 (28.1%) cases were females. Thirty seven cases had cardiovascular events during 6 to 12 months after the study began. The results of the MDR analysis and verification using Chi-square test showed that the development of cardiovascular events was positively correlated with interaction between blood stasis and toxic syndrome at the baseline, blood stasis at the baseline and qi deficiency at the 6-month follow-up, toxic syndrome at the baseline and qi deficiency at the 6-month follow-up, toxic syndrome at the base line and blood stasis at the 6-month follow-up, qi deficiency and blood stasis at the 6-month follow-up (P<0.05 for all).

CONCLUSIONSBlood stasis, toxic syndrome and qi deficiency are important factors of stable CHD. There: are positive correlation between cardiovascular events and syndrome evolution from blood stasis to qi deficiency, from toxic syndrome to qi deficiency and from toxic syndrome to blood stasis, indicating the pathogenesis of toxin consuming qi, toxin leading to blood-stasis in stable CHD patients prone to recurrent cardiovascular events.

Aged ; Cardiovascular Diseases ; etiology ; Coronary Angiography ; Coronary Disease ; complications ; physiopathology ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Multifactor Dimensionality Reduction ; Syndrome

Recent studies have found that ABO blood group antigen is also closely related to the onset and development of many diseases. More and more attention is being paid to the decrease of A/B blood group antigen caused by some tumors. This study was purpose to investigate the correlation between DNA methylation of the ABO gene promoter CpG island and leukemia. The relative contents of ABH antigen on the surface of RBC from kinds of blood disease patients and healthy individuals were detected by using flow cytometry and confocal laser scanning microscopy. The DNA sequences and CpG methylation of ABO gene promoter in patients with hematopathy and healthy individuals, as well as the -102 site methylation of ABO gene promoter in patients with hematopathy and healthy individuals were detected by PCR and MSP-PCR respectively. The results showed that RBC from leukemia patients displayed different degree of A/B antigen decrease. The sequences of ABO gene promotor of patients with hematopathy were not different from healthy individuals indicating high conservation of promoter sequences. Comparison of sequences between patients with hematopathy and healthy individual indicated that CpG islands on ABO gene promoter either from blood disease patients or from healthy individual had no methylated site in AA patients, but C residues at position -102, -101, -100, -99 and -97 on the promoter of ABO gene in AML, CML, ALL and some MDS patients were methylated. It is concluded that methylation of CpG islands in promoter of ABO gene may result in AB antigen decrease in patients with leukemia. The methylation sites -102, -101, -100, -99 and -97 may be specific for leukemia. The methylation of site -102 can be used as a molecular marker in differential diagnosis for leukemias.
ABO Blood-Group System ; genetics ; Base Sequence ; CpG Islands ; genetics ; DNA Methylation ; Humans ; Leukemia ; genetics ; Molecular Sequence Data ; Promoter Regions, Genetic ; Sequence Analysis, DNA

BACKGROUNDCytomegalovirus (CMV) retinitis is the most severe intraocular complication that results in total retinal destruction and loss of visual acuity in patients with acquired immunodeficiency syndrome (AIDS). This study aimed to investigate the fundus characteristics, systemic manifestations and therapeutic outcomes of CMV retinitis associated with AIDS.

METHODSIt was a retrospective case series. CMV retinitis was present in 39 eyes (25 patients). Best corrected visual acuities, anterior segment, fundus features, fundus fluorescence angiography (FFA) and CD4(+) T-lymphocyte counts of the patients with CMV retinitis associated with AIDS were analyzed. Intravitreal injections of ganciclovir (400 µg) were performed in 4 eyes (2 patients).

RESULTSRetinal vasculitis, dense, full-thickness, yellow-white lesions along vascular distribution with irregular granules at the border, and hemorrhage on the retinal surface were present in 28 eyes. The vitreous was clear or mildly opaque. Late stage of the retinopathy was demonstrated in 8 eyes characterized as atrophic retina, sclerotic and attenuated vessels, retinal pigment epithelium (RPE) atrophy, and optic nerve atrophy. Retinal detachment was found in 3 eyes. The average CD4(+) T-lymphocyte count in peripheral blood of the patients with CMV retinitis was (30.6 ± 25.3) × 10(6)/L (range, (0 - 85) × 10(6)/L). After intravitreal injections of ganciclovir, visual acuity was improved and fundus lesions regressed.

CONCLUSIONSCMV retinitis is the most severe and the most common intraocular complication in patients with AIDS. For the patients with yellow-white retinal lesions, hemorrhage and retinal vasculitis without clear cause, human immunodeficiency virus (HIV) serology should be performed. Routine eye examination is also indicated in HIV positive patients.

Acquired Immunodeficiency Syndrome ; complications ; immunology ; metabolism ; Adult ; Antiviral Agents ; pharmacology ; CD4-Positive T-Lymphocytes ; metabolism ; Cytomegalovirus Retinitis ; drug therapy ; etiology ; immunology ; metabolism ; Female ; Fluorescein Angiography ; Ganciclovir ; pharmacology ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult