Objective To study the relation between the HPV6/18 virus infection and the development of pathological changes of cervix. Methods The number of HPV16/18 DNA copies and the expression rate of HPV16/18 E7 mRNA in the pathological cervix were examined by the quantitative fluorescent PCR combined with pathological diagnosis and immunohistochemistry staining. Results The HPV16 infection rates in chronic cervicitis group were much lower (7.4%) than that in the cervical intraepithelial neoplasia (CIN) groups and the cervical cancer group (69.6% and 72.7%), respectively. Statistical analysis showed that the difference of HPV16 DNA copies was not significant between the chronic cervicitis group and CIN groups. In contrast to the above mentioned result, the number of HPV DNA copies between the CIN groups and the cervical cancer group was significantly different. The HPV16 E7 gene expression rates in CIN Ⅰ, Ⅱ, Ⅲ and cervical cancer groups were 0,37.5%,42.9%,63.6%, respectively. Conclusion Ins more common than that with HPV18. The number of HPV16 DNA copies in cervical cancer tissues is markedly higher than that in CIN Ⅱ, Ⅲ groups. The HPV16 E7 mRNA expression is significantly increased in the cervical cancer, and it is more closely correlated to this pathological changes. The quantitative fluorescent PCR can be used to reflect the activity of HPV, and it is a useful method for the screening examination of HPV and for the early diagnosis and treatment of cervical caner.

Objective To explore the clinical characteristics,therapy and prognosis of primary central nervous system lymphoma (PCNSL).Methods Forty-three PCNSL patients were retrospectively analyzed.Results The most frequent PCNSL clinical presentations were an increased intracranial pressure, focal neurologic deficits.The median survival time of patients was 18.0 months (14.2—21.8 months,95% CI).Cox regression model analysis revealed that ependymal and leptomeningeal involvement was the only independent factor for overall survival (P=0.002).Conclusions PCNSL is a aggressive disease with a poor prognosis and difficult to diagnose before operation.The optimal treatment modality for newly diagnosed PCNSL is still under investigation.

Objective Feitai Capsule,a compound of traditional Chinese herbal medicine,has been screened and refined repeatedly for many years and shown to have a good anti-tumor effect.Strict quality control and further screening of the efficacious components of the compound are of great clinical significance.The purpose of this study was to establish the methods for determining the isofraxidin content in Feitai Capsule.Methods We determined the content of isofraxidin in Feitai Capsule by high performance liquid chromatography (HPLC),using the chromatographic column Hypersil ODS-C18 (4.6 mm?150 mm,5 ?m),with the mobile phase as acetonitrile 0.2% phosphoric acid solution (21∶79),the flow rate of 1.0 ml/min,the detective wavelength of 344 nm and the column temperature at 30℃.Results Isofraxidin showed a good linearity,within the range of 2.00-10.80 ?g/ml (y=69 427x+15961,r = 0.999 9),with the average recovery of 97.89% and RSD of 1.64% (n = 6).Conclusion HPLC,accurate and reproducible,is suitable for the determination of the isofraxidin content in Feitai Capsule.

OBJECTIVETo evaluate the clinical efficacy and efficacy sustainable time of acupuncture in knee osteoarthritis (KOA).

METHODSThe non-randomized concurrent control trial was adopted. One hundred and ninety-three cases of KOA were divided into an immediate acupuncture group (group A, 97 cases) and a delayed acupunc-weeks at the end of treatment. In group B, the same acupuncture therapy was applied after waiting 4 weeks. The acupoints in the two groups were Liangqiu (ST 34), Dubi (ST 35), Zusanli (ST 36), Yanglingquan (GB 34), Yinlingquan (SP 9), Xuehai (SP 10), Xiyan (EX-LE 4), Xiyangguan (GB 33). WOMAC (Western Ontario and McMasters Universities Osteoarthritis) was used for the assessment of the primary index and VAS (visual analogue scale) was for the secondary index. The evaluation was accomplished by the patients at the beginning of trial, on the 4th and 8th weeks. In each group, 72 patients finished the trial and the data of the lost cases were included in the final data analysis.

RESULTSIn the 4th week of trial, WOMAC score was (25. 8+/-22.0) in group A difference (P<0. 001). VAS scorewas (31. 8+/-24. and was (43.8+/-22.2) in group B, indicating the significant 6) in group A and was (56. 6 +/-25. 8) in group B, indicating very significant difference (P<0. 001). In the 8th week, the efficacy was reduced slightly in the follow-up of group A, but it was improved apparently as compared Acupuncture relieves joint pain and improves joint function obviously.by th patiĩeffr,a Mtaetfti-?an tf ri-with that before treatment.

CONCLUSIONAcupuncture relieves joint pain and improves joint function obviously.The effect of acupuncture is still sustainable in 4 weeks after terminating the treatment.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Arthralgia ; therapy ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; therapy ; Treatment Outcome

OBJECTIVETo assess the association between leptin gene promoter methylation and serum leptin concentrations in patients with impaired glucose regulation (IGR) and type 2 diabetes mellitus (T2DM).

METHODSMethylation status of leptin gene promoter was determined with methylation-specific polymerase chain reaction. Serum leptin concentrations were determined using enzyme-linked immunosorbent assay.

RESULTSAmong three groups of individuals with different levels of glucose, the methylation rates of leptin gene in IGR and T2DM groups were 43.6 % and 31.5 %, respectively, which were significantly lower than that of healthy subjects (59.2%; Chi-square=22.499 and 5.109, respectively, P<0.05). A lower methylation rate was also observed in T2DM group compared with IGR group (Chi-square=3.962, P<0.05). Leptin levels in both T2DM and IGR groups were elevated compared with normoglycemic subjects, but only T2DM group was significantly higher (q=6.81, P<0.01). Linear regression analysis indicated that serum leptin concentrations has increased along with declining of DNA methylation rate (r=-0.95, P<0.01).

CONCLUSIONLower levels of leptin gene promoter DNA methylation and serum leptin concentrations are associated with the development of diabetes. Measurement of the methylation status of leptin gene promoter and expression can facilitate early intervention of the disease.

DNA Methylation ; Diabetes Mellitus, Type 2 ; genetics ; metabolism ; Female ; Genetic Predisposition to Disease ; Glucose ; genetics ; metabolism ; Humans ; Leptin ; blood ; genetics ; metabolism ; Male ; Middle Aged ; Promoter Regions, Genetic

OBJECTIVETo evaluate the clinical efficacy of the conventional acupuncture on knee osteoarthritis (KOA).

METHODSSeventy-three cases of KOA were treated with the conventional acupuncture therapy at Liang-qiu (ST 34), Dubi (ST 35), Zusanli (ST 36), Yanglingquan (GB 34), Yinlingquan (SP 9), etc. The conventional acupuncture therapy lasted for 4 weeks (at least 3 treatments and less than 5 times each week). At the end of treatment, the follow-up visit lasted for 4 weeks. The Western Ontario and McMaster Universities osteoarthritis index (WOMAC) and the visual analogue scale (VAS) were adopted for the efficacy assessments of the patients.

RESULTSOf 73 cases, 49 cases accomplished the trial. The data of the lost cases were included in the final data analysis. At the end of 4-week treatment and the follow-up visit, every score in VAS and WOMAC was decreased The conventional acupuncture therapy reduces the joint pain for apparently (P < 0.001, P < 0.05).

CONCLUSIONthe patients with KOA and improves the joint function. The 4-week follow-up visit indicates that the efficacy of acupuncture is still sustained.

Acupuncture Therapy ; Aged ; Aged, 80 and over ; Arthralgia ; therapy ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; therapy ; Pain Measurement ; Treatment Outcome

Objective To investigate the diagnostic value of prenatal ultrasonography in the fetal hemivertebra. Methods The ultrasonographic findings of three fetuses with hemivertebra diagnosed in our hospital were reviewed and compared with those of postnatal ultrasonography,other image modalities,and autopsy. Results In all fetuses,a distortion of the spine was observed where only one half of the vertebra could be identified. The parents opted for termination of the pregnancy in one case and the deformity was confirmed by autopsy. The other two fetuses were delivered and in one fetus the diagnosis was confirmed by radiological assessment. Conclusions Hemivertebra can be diagnosed accurately by second-trimester ultrasonography. The prognosis is mostly favorable when no other anomalies are associated. Meticulous examination may disclose the lesion and help decide the fate of pregnancy.

Objective To determine the diagnostic value of two and three dimensional ultrasound in detecting fetal hand malformations. Methods In the retrospectively analysis,the severe fetal hand malformations detected by prenatal ultrasound during the recent three years in our hospital were classified according to the prenatal sonographic characteristics,family history,karyotype analysis and autopsy results,etc.Results Fourteen fetuses with hand-anomaly were detected during the 16-28th week of gestation,both hands were affected in 10 cases,with the same morphology bilaterally in 8 cases.Thirteen cases had other abnormal sonographic findings.They were detected and classified into three categories,Wrist deformity (9 cases),three fetuses were associated with total absence of radius and radial clubhand,and six fetuses had palmar deviated hands,with various etiologies including familial hereditary arthrogryposis multiplex congenita,distal type 1 (AMCD1),amniotic band syndrome,body stalk anomaly,trisomy18 and micromelia.Hand (figer) hypoplasia or aphasia (3 cases),one hand was absent in one fetus without associated anomaly,absence of five fingers with ipsilateral multicystic dysplastic kidney in the second fetus,and the third fetus had split hand/foot malformation (SHFM).Overlapping fingers (4 cases),three of them were trisomy18,and two fetuses had both wrist deformity and overlapping fingers.Conclusions Prenatal two and three dimensional ultrasound play an important role in detecting and diagnosing severe type of fetal hand malformations.

Objective A novel multiplex real-time RT-PCR kit was developed to detect EV71,CoxA16 and other human enteroviruses simultaneously with an internal amplification control to avoids false negatives,which used for hand,foot and mouth disease in the clinical diagnosis and epidemiological surveillance.Methods Design specific primers and probes of EV71,CA16,other intestinal virus and internal amplification control,improve the extraction method of virus nucleic acid.Optimization the detection system of real-time quantitative PCR.Research the products of the accuracy,stability,precision,amplification efficiency and detection of linear range.Results The primers and probes had high spicificity.The Viral RNA extraction effect of this Kit is as same as that of QIAamp Viral RNA mini Kit (QIAGEN company),but less reagent cost.The optimal concentrations of primers and probes are 0.2 μmol/L for all the upstream and downstream primers,0.06 μmol/L for probes of other human enteroviruse,0.08 μmol/Lfor probes of EV71 and CA16 respectively.The kit has good stability,accuracy and precision.The amplification efficiencies of EV71,CoxA16 and other human enteroviruses are 106％,101％ and 105％ and the detection of linear range is from 109 copies/μl-102 copies/μl.Conclusion The novel multiplex realtime RT-PCR kit for detecting EV71,CoxA16 and other human enteroviruses simultaneously with an internal amplification control has good stability,accuracy,precision and amplification efficiencies.So it has great value in clinical application.

OBJECTIVETo prepare pneumolysin as a new protein carrier of vaccine against otitis media with genetic engineering technology and establish the base of the study on pneumococcal conjugative vaccines.

METHODSGenomic DNA was isolated from streptococcus pneumoniae. A pair of primers which included two restriction sites was designed based on the published pneumolysin gene sequence. The pneumolysin gene was amplified from pneumococcal DNA with PCR technology. The restriction enzyme digested fragment was linked into the cloning vector PET-28a and the recombinant plasmid DNA containing pneumolysin was then transfected into host cell E. coli JM109 (DE3).

RESULTSDNA fragments were subcloned to construct the complete pneumolysin gene by a conventional coning and PCR. The inserted pneumolysin gene sequence was confirmed by DNA sequencing and the pneumolysin protein was successfully expressed. The relative molecular mass of the expressed product was 52 000. The expressed product amounted to 8% of the total host cell protein.

CONCLUSIONSThe pneumolysin gene was successfully cloned into host cell using genetic engineering technology. The recombinant pneumolysin was expressed and purified for preparation. This work laid a foundation of the preparation of pneumococcal conjugative vaccines.

Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Genetic Engineering ; Genetic Vectors ; Plasmids ; Pneumococcal Vaccines ; genetics ; Streptococcus pneumoniae ; genetics ; Streptolysins ; biosynthesis ; genetics