OBJECTIVE: To establish optimal amplification conditions with the application of 0.2 mL test tube in single cell separation and inspection. METHODS: Oral epithelium cell suspension was prepared. Five or ten cells were collected with 0.2 mL test tube. Then DNA were amplified with Identifiler Plus kit in three different conditions in which the proteinase K addition, gold enzyme concentration in PCR reaction, and PCR reaction cycles were adjusted separately. Finally the detection rate, allelic dropout rate and artificial alleles were compared among the groups. RESULTS: In these 3 different conditions: addition of proteinase K, addition of 0.4 microL gold enzyme in PCR reaction, and use of 32 cycles, the detection rate was higher and allelic dropout rate was lower than the other conditions. CONCLUSION In single cell separation and inspection, the usage of 0.2 mL test tube could be an effective supplement to chip-low volume amplification.
Alleles ; Cell Separation/methods* ; DNA/genetics* ; DNA Fingerprinting/methods* ; Endopeptidase K/administration & dosage* ; Enzymes/administration & dosage* ; Epithelial Cells/cytology* ; Feasibility Studies ; Forensic Genetics ; Genotype ; Humans ; Mouth Mucosa/cytology* ; Nucleic Acid Amplification Techniques ; Polymerase Chain Reaction/methods* ; Sensitivity and Specificity

OBJECTIVETo evaluate the efficacy of Cotrel-Dubeusset (CD) instrumentation combined with translaminar facet joint screw (TLS) in the treatment of thoracolumbar fracture.

METHODSA total of six L(2)-L(4) spines were used to establish unstable fracture model with three-dimensional range of motion (ROM) of the spines measured. Fixation with CD and fixation with CD combined with translaminar facet joint screw were achieved to compare their stability. Thirty cases of thoracolumbar fracture, in whom the anterior edge of vertebral body was compressed to 59% and the posterior edge compressed to 88%, were treated by pedicle screw fixation combined with TLS. Among them, 19 received posterolateral or anterior-posterior bone grafting.

RESULTSThere was significant difference in ROM between the two techniques except that in extension. In Group CD+TLS, ROM was 5.38% lower, lateral bending 4.91% lower and axial rotation 11.85% lower than those in Group CD respectively. In the clinical group, the average anterior edge restored to 97% and posterior edge to 98%. The duration of follow-up was 5-24 months (mean, 10 months). The rate of correction loss on the anterior edge was 4.5%. Among the 19 cases of bone grafting, all of them achieved bony fusion (mean fusion time, 4.3 month) with a correction loss rate of 3.4%.

CONCLUSIONSIn the treatment of thoracolumbar fracture, pedicle screw fixation combined with TLS can strengthen the stability of pedicle screws, especially anti-rotation stability and enhance fusion rate and reduce correction loss.

Adult ; Bone Screws ; Bone Transplantation ; Female ; Fracture Fixation, Internal ; instrumentation ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Range of Motion, Articular ; Spinal Fractures ; physiopathology ; surgery ; Spinal Fusion ; instrumentation ; methods ; Thoracic Vertebrae ; injuries

Objective To retrospectively analyze the significance of dynamic intracranial pressure monitoring and routine monitoring in the treatment of severe traumatic brain injury.Methods Forty-two patients with severe craniocerebral trauma who were admitted into our hospital from March 2013 to December 2015 and underwent intracranial pressure monitoring were enrolled in this study as the observation group.Thirty-nine patients with severe traumatic brain injury who were routinely monitored within 3 hours after admission were selected as the control group in the corresponding period.Timely take drugs or surgical treatment according to the monitoring results,and analyzed the clinical efficacy,craniotomy cases,time of admission to craniotomy,and complications of the two groups.Results The cases with good prognosis in the control group was 24 (61.5％) while it was 31 (73.8％) in the observation group,and the difference was statistically significant (P ＜ 0.05).The cases with poor prognosis in the control group was 15 (38.5％) while it was 11 (26.2％) in the observation group,and the difference was statistically significant(P ＜0.05).Therer were 13 cases (30.1％) of craniotomy in the control group and 5 cases (12.8％) in the observation group with statistically significant difference (P ＜ 0.05).The time of admission to craniotomy in the control group was (24.5 ± 1.7) hours,and it was (18.3 ± 2.4) house in the observation group with statistically significant difference (P ＜ 0.05).The incidence of intracranial infection complication was 9.5％ in the control group and 8％ in the observation group.There was no significant difference between the two groups (P ＞ 0.05).Conclusion Invasive intracranial pressure monitoring can reflect the changes of patients in time,which can improve the clinical curative effect and would not increase the incidence of intracranial infection.

Objective To investigate the effect of change in neurotrophins level in the bronchial asthma rats' lung on airway hyperresponsiveness and airway neural plasticity. Methods A total of forty male SD rats were randomly divided into 4 groups with 10 rats in each group: control group,asthmatic group, NGF+BDNF prevention group and anti-NGF+anti-BDNF prevention group. The asthmatic model was established by inhalation and injection of ovalbumin. The airway responsiveness was measured after 8 weeks.The bronchial inflammation was assessed by HE staining,and nerve growth factor and brain derived neurotrophic factor expressions of left lung were assayed by the immunohistochemistry staining.Then the expressions of synaptophysin and neurofilament were detected by RT-PCR. Results Compared with the control group, the lung tissue of the asthma group and NGF+BDNF prevention group had more infiltrating inflammatory cells; The expressions of NGF and BDNF were higher in the asthma group and NGF+BDNF prevention group than those in the control group and anti-NGF+anti-BDNF prevention group (P＜0.05),and significantly higher in the NGF+BDNF prevention group than those in the asthma group (P＜0.05).Both the airway responsiveness and the levels of SYN mRNA and NF mRNA in the lung tissues were significantly higher in the asthma group and NGF+BDNF prevention group than those in the control group (P＜0.05).In asthma group,the expressions of NGF and BDNF were positively related to the expressions of SYN (r=0.889,P＜0.05; r=0.985,P＜0.05)and NF(r=0.956,P＜0.05; r=0.927,P＜0.05),and also positively related to the airway hyperresponsiveness (r=0.938,P＜0.05; r=0.906,P＜0.05). Conclusion NGF and BDNF might be involved in rat airway bronchial neural plasticity changes,which resulted in the airway hyperresponsiveness.

OBJECTIVETo investigate the clinical features, pathology, diagnosis and treatment of inverted urothelial papilloma.

METHODSA total of 151 cases of urothelial inverted papilloma were analysed retrospectively. Of the cases, 134 were male and 17 were female, with a mean age of 54 years old. Most patients complained of painless gross hematuria. The diagnosis could be established mainly by ultrasonic, intravenous urography, retrograde pyelography, cystoscope and pathology. Among them, 7 cases who had the papilloma at upper urinary tract underwent nephroureterectomy except one. One hundred and forty-four cases had the papilloma at low urinary tract, with 124 treated by transurethral bladder tumor resection (TURBT), among which 11 cases accompanying benign prostatic hyperplasia were treated by transurethral prostatic resection, 3 by transurethral resection of prostatic urethral tumor, 15 by partial cystectomy, 2 by total cystectomy.

RESULTSOne hundred and eighteen cases were followed up 1 year to 12.5 years (mean 6.3 years). Intravesical recurrence was found in 5 cases. Of them 2 cases developed malignance in 8 and 30 months postoperatively, and 1 case underwent total cystectomy.

CONCLUSIONSInverted urothelial papilloma is a benign tumor, which appears male predominant. Most of the lesions are found in the bladder. TURBT is the preferred treatment choice for inverted papilloma of the bladder. Although this disease has a good prognosis, regular follow-up observations are necessary.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Papilloma, Inverted ; diagnosis ; pathology ; surgery ; Retrospective Studies ; Urologic Neoplasms ; diagnosis ; pathology ; surgery

OBJECTIVETo investigate the human mitochondrial DNA (mtDNA) variations associated with longevity in Bama elderly population from Guangxi.

METHODSMitochondrial genome of 20 individuals over 96 years of age was sequenced, and seven target single nucleotide polymorphism (SNPs) were observed by comparing with the standard rCRS sequence, and two were tested by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in a larger population including 208 individuals of 90-113 years old, and 586 unrelated control individuals from Guangxi.

RESULTSThe 4824G frequency of the mtDNA4824A/G locus increased with age both in the long-lived elderly and in controls. And it was significantly higher in controls than that in long-lived population (P<0.05).

CONCLUSIONThe mtDNA4824 A/G is not only an age-related locus, its mutation is also negatively correlated with longevity.

Aged ; China ; ethnology ; DNA, Mitochondrial ; analysis ; genetics ; Genome, Mitochondrial ; genetics ; Haplotypes ; Humans ; Longevity ; genetics ; Mutation ; Myanmar ; ethnology ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; genetics ; Population Groups

Objective To evaluate interventional therapy for biliary stricture (BS) after orthotopic liver transplantation (OLT). Methods The efficacy of interventional therapy for BS after OLT from Oct 2003 to Jan 2006 was analyzed retrospectively. Fifty-three patients received 107 times of interventional therapy through endoscopic retrograde cholangiography ( ERC) which included 68 nasobiliary catheter placements,26 biliary balloon dilatations and stent placements and 13 ERC. Nine patients received 11 times of interventional therapy through percutaneous transhepatic cholangiography ( PTC) including 2 PTC, 7 percutaneous drainages,3 biliary balloon dilatations and 1 biliary stent replacement. One patient received bile drainage through T tube. Results The success rate of ERC was 88. 8% (95/107) , that of nasobiliary catheter placement 94% (64/68) , biliary stent placement 88. 5% (23/26). The success rate of PTC was 81. 8% (9/11) , that of percutaneous drainage was 100% (7/7) , biliary stent replacement 100% (1/1). The curative rate of interventional therapy for 53 patients with BS was 28. 3% (15/53) ,the improvement rate was 41. 5% (22/53). The curative rate of interventional therapy for anastomotic, extrahepatic, intrahepatic hilar and diffuse BS was respectively 66. 7% (4/6)、66. 7% (10/15)、50% (1/2)、0 (0/7) and 0 (0/22). Conclusions The efficacy of interventional therapy for BS after OLT was not satisfactory. The result relates to the type of BS, for anastomotic, extrahepatic and solitary intrahepatic BS this therapy was effective, while that for hilar and diffuse BS the prognosis was poor.

Objective To report initial experience and the indications of retroperitoneal laparoscop- ic partial nephrectomy.Methods From December 2001 to October 2005,23 patients (including 14 cases of renal cell carcinoma,5 of hamartoma and 4 of duplex kidney) underwent retroperitoneal laparoscopic par- tial nephrectomy.One of the patients had solitary kidney.Results All the operations were successful ex- cept for only 1 requiring conversion to laparoscopic nephrectomy because of bleeding resulting from blocking anterior branch of renal artery.The mean operative time was 121 min (60-240 min),and mean ischemic time was 32 min (20-55 min).The estimated bleeding volume ranged from 100 ml to 300 ml,and no pa- tient needed blood transfusion.Pathology showed negative surgical margins in 14 cases of renal cell carcinoma and confirmed the diagnosis of hamartoma in 5 cases.One patient with duplex kidney required open partial nephrectomy because of renal cystic mass resulting from incomplete resection.Conclusions Retroperito- neal laparoscopic partial nephrectomy offers a new effective and minimally invasive treatment for selected pa- tients with renal mass.The long-term effects of the procedure need further investigation.

Ischaemic stroke is a common condition leading to human disability and death. Previous studies have shown that oleanolic acid (OA) ameliorates oxidative injury and cerebral ischaemic damage, and miR-186-5p is verified to be elevated in serum from ischaemic stroke patients. Herein, we investigated whether OA regulates miR-186-5p expression to control neuroglobin (Ngb) levels, thereby inhibiting neuronal pyroptosis in ischaemic stroke. Three concentrations of OA (0.5, 2, or 8 μM) were added to primary hippocampal neurons subjected to oxygen–glucose deprivation/ reperfusion (OGD/R), a cell model of ischaemic stroke. We found that OA treatment markedly inhibited pyroptosis. qRT–PCR and western blot revealed that OA suppressed the expression of pyroptosis-associated genes. Furthermore, OA inhibited LDH and proinflammatory cytokine release. In addition, miR-186-5p was downregulated while Ngb was upregulated in OA-treated OGD/R neurons. MiR-186-5p knockdown repressed OGD/R-induced pyroptosis and suppressed LDH and inflammatory cytokine release. In addition, a dual luciferase reporter assay confirmed that miR-186-5p directly targeted Ngb. OA reduced miR-186-5p to regulate Ngb levels, thereby inhibiting pyroptosis in both OGD/R-treated neurons and MCAO mice. In conclusion, OA alleviates pyroptosis in vivo and in vitro by downregulating miR-186-5p and upregulating Ngb expression, which provides a novel theoretical basis illustrating that OA can be considered a drug for ischaemic stroke.

OBJECTIVETo analyze the distribution features of Gleason score and evaluate the relationship between Gleason score and clinical stages in patients with prostate cancer.

METHODSSurveys were made of the inpatients with prostate cancer diagnosed by pathology from January 1992 to June 2005 in our hospital. Gleason score and clinical stages were determined on the basis of pathological examination and clinical data of the prostate cancer patients. The patients were divided into three groups (1992-1999, 2000-2002 and 2003-2005). The Chi-square test was used to evaluate the distribution and differences of Gleason score among the three groups. Spearman rank correlation was applied to the evaluation of the relationship between Gleason score and clinical stages.

RESULTSWe found a statistically significant shift in the distribution of Gleason score (chi2 = 17.703, P < 0.01), and a slight increase in the mean Gleason score. The proportion of moderately differentiated tumor increased (chi2 = 10.736, P < 0.01). There was little change in the proportion of Gleason score 7, 8, 9 and 10 (chi2 = 4.038, P > 0.05). Gleason score had a significant positive correlation with clinical stages in the 346 cases of prostate cancer (r = 0.452, P < 0.01). Significant difference was observed between Gleason score 2-6 and 7 or 8-10 (chi2 = 8.786, P < 0.01, chi2 = 22.956, P < 0.01), but not between the latter 2 groups (chi2 = 0.787, P > 0.05) in prediction of organ-confined disease.

CONCLUSIONSGleason score 7 shows the similar value to Gleason score 8-10 in predicting the progression of the disease. Gleason score was significantly correlated with clinical stages, which suggests that Gleason score is also an important indicator for the prognosis of prostate cancer.

Adult ; Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Prostatic Neoplasms ; pathology ; Retrospective Studies