Objective To resolve the problem of the accuracy and standardization of STR-PCR typing in forensic practice,construct DXS6804 allelic ladder by molecular clonning and apply them in a population study on the Pumi population in Yunnan,China.Methods PCR was used to produce several different allelic fragments of the locus.After cloning the PCR products,the recombinant plasmids were sequenced.Then we denominated them and used them as template for re-amplification to generate the locus standard ladder.Results The sequencing results confirmed that the size and the construction of the inserts were correct.The genetic polymorphisms of this locus in Yunnan Pumi population of China were studied.Two off-ladder alleles of DXS6804 locus were found.Conclusion This method is of high value for forensic DNA typing to construct standard ladders.DXS6804 is robust for genetic research and forensic application.

Brain Ischemia ; etiology ; Child ; Humans ; Male ; Metabolic Syndrome ; complications ; Stroke ; etiology

Objective To investigate the genetic polymorphism of DXS8378 STR locus of chromosome X in Chinese Lisu,Pumi and De'ang populations in Yunnan and construct relative standard allelic ladders.Methods After being amplified by PCR,different STR allelic fragments were isolated from the PAG electrophoresis.The STR allelic fragments were extracted by kit and reamplified by PCR to obtain purified allelic fragments.Next,the purified allelic fragments were subcloned individually into the PUC plasmid vectors,and the size and structure of the inserts were confirmed by the analysis of their DNA sequences.Then we transfected it into competent E.coli DH5?TM cells,and finally,the recombinant plasmids DNA with the inserts were used as template for reamplification to generate the standard ladders.Results The standard allelic ladder for DXS8378 locus was obtained,with which the genetic polymorphisms of DXS8378 locus in three Chinese populations in Yunnan were studied.Conclusion The standard ladder made by this method is excellent,and DXS8378 is powerful for forensic practice in Chinese population.

Objective To learn about electrolytes imbalance and water intoxication in children treated with high-dose cyclophosphamide(HD-CTX)as well as the renal function and the relative clinical symptoms,and study the mechanisms of hyponatremia.Methods Patients' clinical manifestations during and after HD-CTX therapy were summarized.Serum electrolytes and creatinine(Cr)were detected before and 6 or 8 hours after therapy with HD-CTX,antidiuretic hormone(ADH) in some patients were measured.Results Of 108 therapeutic cases 24 accompanied with vomits and 22 with a decreased urine output,in which 4 developed eyelid or ankle edema.Seven cases had neural-sarcous symptoms and 5 cases had abdominal pain or diarrhea.Serum sodium decreased significantly after HD-CTX[(139.12?3.30) mmol/L vs(134.06?8.23) mmol/L] in whom rechecked after 6 h,(141.77?3.59) mmol/L vs(133.26?6.41) mmol/L in those rechecked after 8 h(Pa0.05].Serum Cr increased 8 h after therapy[(29.95?13.61) ?mol/L vs(43.33 ? 17.25) ?mol/L P

Objective To develop a rapid and sensitive DNA microarray for Listeria monocytogenes detection.Methods A DNA microarray was developed using gyrB,ISR,16S rRNA,23S rRNA,hlyA,iap and prfA as the target genes and tested against 18 different species of known reference for repeatability,sensitivity,and specificity to verify the effectiveness of the chip.Results After testing of samples by the LM array,results show that the 70 mer Oligos synthesized by IDT are superior to the Oligos synthesized by Sagon with respect to both probe spotting or samples detection.The comparison of 3 spotting probe concentrations of 10 μmol/L,40 μmol/L and 80 μmol/L demonstrated that the 10 pmol/L probes result in good detection signals equivalent to the 40 μmol/L and 80 μmol/L probes.The repeatability and sensitivity evaluated by sample testing on the LM array revealed that the chips developed in this study have good repeatability and the lower limit of sample detection is 0.9 ng DNA.The LM array can distinguish clearly and definitively between Listeria and non-Listeria bacteria in the sample.Conclusion The microarray is able to rapidly detect and identify Listeria monocytogenes.

OBJECTIVE: To study the association of the vitamin D receptor gene BSM Ⅰ, TAQ Ⅰ and APA Ⅰ genetic polymorphisms with bone mineral density and biochemical markers of bone turnover in postmenopausal women.METHODS: ①total of 576 postmenopausal Han ethnic women of 48-84 (62.17±6.37) years old in Fuzhou city were investigated, on the basis of their informed consent, through random sampling method from January 2007 to December 2008. ②The subjects were recorded regarding to their age, menopause duration, body mineral index and postmenopausal fracture incidence. ③Dual energy X-ray absorptiometry was used for measuring the bone mineral density of vertebrae L<,2-4>, left femoral neck, trochanter and Ward's triangle. ④The genetic polymorphisms of vitamin D receptor gene BSM Ⅰ, TAQ Ⅰ and APA Ⅰ were detected using polymerase chain reaction-rastriction and fragment length polymorphism (PCR-RFLP) technique. ⑤The biochemical markers of bone turnover (serum bone gla protein, serum bone alkaline phosphatase, urinary pyddinoline and urinary deoxypyridinoline) were detected with enzyme linked immunosorbent assay.RESULTS: A total of 561 subjects up to standard were involved in the result analysis. ①There was no significant difference in bone mineral density among genotypes of vitamin D receptor gene BSM Ⅰ, TAQ Ⅰ and APA Ⅰ polymorphisms (P > 0.05). ②There was no significant difference in the biochemical markers of bone tumover among genotypes of BSM Ⅰ, TAQ Ⅰ and APA Ⅰ polymorphisms (P > 0.05). ③There was no significant difference in the incidence of osteoporosis among genotypes of BSM Ⅰ, TAQ Ⅰ and APA Ⅰ polymorphisms (P > 0.05). ④There was no significant difference in the incidence of postmenopausal fracture among genotypes of BSM Ⅰ, TAQ Ⅰ and APA Ⅰ polymorphisms (P > 0,05).CONCLUSION: BSM Ⅰ, TAQ Ⅰ and APA Ⅰ polymorphisms of the vitamin D receptor gene are not obviously associated with osteoporesis in postmenopausal women, and accordingly can not be taken as genetic markers of osteoporosis in postmenopausal women in Fuzhou.

Objective To investigate the correlation between bone mineral density(BMD)and the PXh haplotype combining estrogen receptor (ER) gene Xba Ⅰ , Pvu Ⅱ polymorphisms and osteocalcin gene Hind Ⅲ polymorphism in postmenopausal women.Methods In 307 subjects,the BMD of lumbar vertebrae and proximal femur were measured by dual energy X-ray absorptiometry and the Xba Ⅰ and Pvu Ⅱ polymorphisms of ER gene and the Hind Ⅲ potymorphism of osteocalcin gene were detected by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP).Results (1)The BMD of greater trochanter was significantly lower in XX genotype group than in xx genotype group ( P<0.05).The BMD of femoral neck, greater trochanter and Ward's triangle were lower in Xx genotype group[(0.695±0.087)g/cm2 , (0.592±0.106)g/cm2, (0.500±0.115) g/cm2] and X allele group[(0.697±0.088)g/cm2 , (0.594±0.105)g/cm2, (0.505±0.123)g/cm2] than in xx genotype group[(0.737±0.108) g/cm2,(0.653±0.119)g/cm2 ,(0.554±0.130)g/cM2] and non-X allele group[(0.737 ± 0.108) g/CM2, (0.653 ± 0.119) g/cm2 , (0.554 ± 0.130) g/cm2] ,respectively (all P<0.05 ).(2)The BMD of Ward's triangle was lower in PP genotype group and P allele group than in pp genotype group and non-P allele group (P<0.05).(3)The BMD of femoral neck, greater trochanter and Ward's triangle were lower in hh genotype group and h allele group than in I-IH genotype group, and were lower in non-h allele group than in HH genotype group(all P<0.05).(4)Women carrying PX, PXh haplotypes combining ER gene and osteocalcin gene had lower BMD at femoral neck than those not carrying PX,not carrying PXh haplotypes, respectively (all P<0.05).ConclusionsER gene(Xba Ⅰ) polymorphism and osteocalein gene(Hind Ⅲ) polymorphism are associated with BMD in postmenopausal women.The presence of X allele or h allele　shows negative influence on the BMD of postmenopausal women.The PXh haplotype is a suitable　genetic marker of postmenopausal women osteoporosis in Fuzhou area.

Objective To investigate the relationship between serum levels of alanine aminotransferase (ALT)or aspartate aminotransferase (AST)apportioned by the same hepatic parenchyma cell volume and liver histological necroinflammation grades in HBeAg-negative chronic hepatitis B (CHB)patients.Methods A total of 145 CHB patients were divided into four groups:Gl,G2,G3 and G4 based on the liver histological necroinflammation grade.The serum ALT and AST levels were determined by automatic biochemical instrument in these four groups.Furthermore,serum ALT and AST levels were then apportioned by the same hepatic parenchyma cell volume.The data were analyzed by ANOVA.Results Mean serum ALT levels in G1,G2,G3 and G4 groups were (35.3±29.1),(91.6±120.4),(111.6± 116.1)and (118.0±122.1)U/L,respectively,and the serum ALT levels apportioned by same hepatic parenchyma cell volume were ( 54.0 ± 45.1 ),( 144.2 ± 184.9 ),(191.3± 204.8)and (215.1 ± 226.5)U/L,respectively.The pairwise comparison between G1 and other three groups all showed statistically significant difference (P＜0.05).Meanwhile,AST levels in G1 to G4 groups were (35.5± 29.0),(64.9±71.7),(96.0±81.9)and (102.8±77.0)U/L,respectively and the serum AST levels apportioned by the same hepatic parenchyma cell volume were (54.3±44.6),(102.3± 107.9),(165.2±148.7)and (189.4±145.4)U/L,respectively.The pairwise comparison between G1 and G3,G1 and G4,G2 and G3,G2 and G4 all showed statistically significant difference (P＜0.05).Conclusion Both AST and ALT levels are sensitive indicators for liver inflammation grading in HBeAg-negative CHB patients during the natural history of the disease.

Objective To evaluate the short-term efficacy of lamivudine versus entecavir for patients with HBeAg-negative acute-on-chronic liver failure (ACLF) with different pretreatment liver failure degrees.MethodsA total of patients with HBeAg-negative ACLF were enrolled into this retrospective study.Seventy-two cases were treated with lamivudine 100 mg daily,while 93 cases were treated with entecavir 0.5 mg daily.Biochemical items,model for end-stage liver disease (MELD)score,hepatitis B virus (HBV) DNA level and mortality were observed.The efficacies of the two drugs were analyzed in patients with different degrees of liver failure.The comparison of rates was done using chi-square test and the measurement data were compared by t test.ResultsAmong the patients with pretreatment MELD scores above 30,the post-treatment HBV DNA levels in lamivudine group and entecavir group were (3.6 ± 1.1) lg copy/mL and (3.7 ± 1.4) lg copy/mL,respectively (t=0.181,P=0.859) and the mortalities were 92.0％ and 91.8％,respectively (χ2 =0.002,P=0.680).For the patients with pretreatment MELD scores from 23 to 30,the post-treatment HBV DNA levels in two groups were (3.2± 1.1) lg copy/mL and (3.2±2.3) lg copy/mL,respectively (t=0.760,P=0.455) and the mortalities were 42.9％,54.1％,respectively (χ2 =0.799,P=0.455).In patients with pretreatment MELD scores below 23,the post-treatment HBV DNA levels in two groups were (3.1±1.0) lg copy/mL and (2.8±1.5) lg copy/mL,respectively (t=-0.740,P=0.464) and the mortalities were 3/19 and 6.3％,respectively (χ2=1.227,P=0.455).In lamivudine group,the mortalities were significantly different among patients with three different ranges of pretreatment MELD scores (χ2 =26.967,P =0.000).The similar differences were also found in entecavir group (χ2 =41.260,P=0.000).ConclusionsAmong treatment na?ve patients with HBeAg-negative ACLF,the short-term efficacy of lamivudine versus entecavir is equal if the degree of pretreatment liver failure is similar.Meanwhile,the degrees of pretreatment liver failure significantly affects the outcome of the treatment.

Objective To explore the diagnostic significance of differential cell count in induced sputum to chronic cough and assessment of airway inflammation.Methods The sputum of 335 chronic cough patients were induced.Differential cell counts were measured in these samples.The side effects were observed during the induced procedure.The final diagnosis was made based on clinical manifestation and examination findings including pulmonary function tests,provocation test,induced sputum cell differentials, etc.Results The cause of chronic cough was defined in 322 patients.The six most important causes of cough were typical asthma(TA,n=84),eosinophilic bronchitis (EB,n=62),atopic cough (AC,n= 42),cough variant asthma (CVA,n=40),gastroesophageal reflux cough(GERC,n=37),rhinitis and/ or paranasal sinusitis (PNDs,n=32),and others and indefinite cause (n=25,13).Percentage of eosinophils were significantly increased in the induced sputum of AC,EB,CVA,and GERC patients (0.005,0.052,0.059,0.234) compared with those in other causes and the healthy controls (0) (P