Objective In this research with the method of Alu -PCR we investigate the integration of hepatitis B virus ( HBV) DNA in human hepatocarcinoma cell line (PLC/PRF/5) genome DNA.Methods We at first extracted the genome DNA from PLC/PRF/5 cells, and then the potential integration fragments of HBV DNA and human genome DNA were amplified with according to the Alu -PCR after three rounds PCR .The Alu-PCR amplification products were observated with agarose gel electrophoresis , then integration positive electrophoresis bandings were chipped and purified for nucleic acid sequencing .At last he bioinformatics information was acquired by blast online.Results Through agarose gel electrophoresis after Alu -PCR amplification, we got four potential integration bindings , among which we got three integration sequences of HBV DNA in human genome DNA .These integration sequences could be individually located in the human chromosome of 03p21.31, 05p15.33, 12q13 and 12-q14.1.Conclusion With Alu-PCR we can accurately measure the integration of HBV DNA in human genome DNA , and Alu-PCR can be a a convenience and economic method in the study of HBV DNA ′s integration in human genome DNA .

Objective To assess the effect of opiod pretreatment on etomidate induced myoclonus. Methods The pertinent literatures were searched by two independent investigators from the following electronic databases:PubMed,Cochrane Library,EMbase,VIP,WanFang Data,and CHKD. Then the meta?analysis was performed by using RevMan 5.2 and STATA 12.0 software. Results A total of 24 RCTs involving 2 396 pa?tients were included for the study. Pretreatment ofμorκ?receptor agonists reduced myoclonus with RR=0.19(95%CI 0.14 to 0.27)and RR=0.22 (95%CI 0.12 to 0.40),respectively. Conclusion Pretreatment of opiods can reduce the incidence of etomidate induced myoclonus.

Objective To evaluate the effects of dexmedetomidine on the hemodynamics and the heart rate variability(HRV)in patients under-going lower limbs operations with application of tourniquet. Methods Forty patients undergoing lower limbs operations with application of tourni-quet were randomized assigned to dexmedetomidine group(group D,n=20)or control group(group C,n=20). After combined spinal-epidural anesthesia,group D received a continuous infusion of dexmedetomidine(0.5μg/kg for 10 min for loading dose and followed by 0.2μg·kg-1·h-1) until tourniquet deflation. The control group received normal saline instead. Mean arterial pressure(MAP),heart rate(HR),saturation of pulse ox-imetry(SpO2),low frequency power(LF),high frequency power(HF)and LF to HF ratio(LF/HF)were recorded at regular time points:imme-diately before loading dose(T0),before tourniquet inflation(T1),15 min after tourniquet inflation(T2),30 min after tourniquet inflation(T3),45 min after tourniquet inflation(T4),60 min after tourniquet inflation(T5),1 min after tourniquet deflation(T6),5 min after tourniquet deflation (T7)and 10 min after tourniquet deflation(T8). Results Compared with T0,the MAP of group D significantly decreased at T6-T8(P<0.05). Compared with T0 and T1,the MAP of group C increased at T2-T5(P>0.05). Compared with T2-T5,the MAP of group C significantly decreased at T6(P<0.05). Compared with group C,the MAP of group D significantly decreased at T6 and T7(P<0.05). Compared with T0,the HR of group D significantly decreased at T1-T8(P<0.05). Compared with T0,the HR of group C had no significant change at T1-T5(P<0.05). Compared with T1-T5,the HR of group D and group C significantly increased at T6(P<0.05). Compared with group C,the HR of group D significantly decreased at T1-T4 and T6(P<0.05). Compared with T0,the SpO2 of group D and group C significantly decreased at T6(P<0.05). Compared with group C,the SpO2 of group D significantly decreased at T1-T3(P<0.05). Compared with T6,LF of group D and group C significantly increased at T7(P<0.05). LF were comparable between groups D and C(P>0.05). Compared with T0,the HF of group D significantly increased and the LF/HF of group D significantly decreased at T1-T4(P<0.05). Compared with group C,the HF of group D significantly increased and the LF/HF of group D significantly decreased at T1-T4(P<0.05). Conclusion The appropriate dose of dexmedetomidine(loading dose 0.5μg/kg and maintenance dose 0.2μg · kg-1 · h-1)can significantly increase vagal tone and improve cardiac sympathetic and parasympathetic balance during tourniquet appli-cation.

Objective To evaluate the profile of transient ischemic attacks in younger patients compared with older patients effectively.Methods We study 75 younger patients(≤45 years)compared with 90 older patients(≥65 years).History of presenting transient ischemic attacks,etiology,clinical feature and lab investigation were compared on the basis of the above age groups.Results Overweight、hypercholesterolemia、insomnia and vascular disease family history being more common in the younger group and Hypertension,ischemic heart disease,long-term history of smoking being more common in the elder group.Diabetes and high serum uric acid occurred high frequently in both groups.Conclusion A significant correlation existed between etiologies contribution and adverse life custom,the early management should be take up to prevent TIA occurrence.

Objective To study the effectiveness of the laboratory diagnosis of multiple myeloma(MM)patients with immun-ofixtion electrophoresis (IFE),protein electrophoresis (SPE)and immunoglobulins and light chain quantitative analysis. Methods Selected 192 MM patients and 30 healthy controls during June 2012 to December 2013 and analyzed the results of IFE,SPE and immunoglobulins,and light chain quantitative analysis in MM patients.Results M protein bands were seen in 120 cases (62.5%)by using SPE and M protein were positive in 174 cases (90.6%)among the 192 MM patients by using IFE.IFE showed that IgG were maximum type of the M protein (106 of 192,55.2%).There were IgG-λ type 66 cases (34.4%),IgA type 36 cases (18.8%)and free light chain type 24 cases (12.5%).Immunoglobulins of different immuno-phenotypes had higher than the nomal group with serum immunoglobulin and light chain quantitative analysis (P <0.05). The detection rate was 67.7% (130/192).Whateverκ-type M protein orλ-type M protein,the ratio ofκ/λwas significantly abnormal (P <0.05).The detection rate was 85.4% (164/192).Conclusion The better detection rate of immunological techniques such as immunofixtion electrophoresis and immunoglobulins quantitative analysis might provide valuable basis for the diagnosis and treatment of MM clinically and prevent misdiagnosis.

Abstract BACKGROUND: Sodium hyaluronate is an effective treatment for osteoarthritis, but the underlying mechanism remains unclear. There is evidence that abnormal expressions of matrix metaloproteinase (MMP)-1, -3 and -9 and tissue inhibitor of metaloproteinase (TIMP)-1 and -2 show great effects on osteoarthritis. OBJECTIVE: To assess the influence of intra-articular injection of sodium hyaluronate on expressions of MMPs-1, 3, 9 and tissue inhibitor of TIMPs-1, 2 in the rabbit cartilage after osteoarthritis. METHODS: Twenty-four mature New Zealand white rabbits were divided into normal control, model, and sodium hyaluronate groups. The model and sodium hyaluronate groups underwent unilateral anterior cruciate ligament transection, and rabbits in the sodium hyaluronate group received 0.3 mL of 1% sodium hyaluronate via intra-articular injection at 4 weeks after modeling, once a week for 5 weeks. At 11 weeks folowing surgery, the rabbits were kiled and the cartilage was harvested to extract total RNA. mRNA expressions of MMPs-1, 3, 9 and TIMPs-1, 2 in the cartilage were analyzed using real-time PCR for each group. RESULTS AND CONCLUSION:Compared with the model group, the range and extent of cartilage damage was reduced in the sodium hyaluronate group (P < 0.01), and Mankin scores were noticeably decreased (P < 0.05). In the cartilage, mRNA expressions of MMPs-1, 3, 9 were enhanced and mRNA expressions of TIMPs-1, 2 were down-regulated in the model group. However, the mRNA expression levels of MMPs-1, 3, 9 and TIMPs-1, 2 in the articular cartilage were not obviously changed in the sodium hyaluronate group. These results suggest that MMPs-1, 3, 9 and TIMPs-1, 2 are involved in the progression of osteoarthritis and the therapeutic mechanism of sodium hyaluronate is not realized through the down-regulation of their expressions during development of osteoarthritis. Sodium hyaluronate for treatment of osteoarthritis is a complex process and the underlying mechanisms require further investigation.

Objective To investigate the clinical significance of homocysteine(Hcy) and glycated hemoglobin (HbA1c) in patients with diabetic retinopathy.Methods From March 2013 to February 2015,a total of 168 patients with diabetes from Beijing Jishuitern Hospital were divided into diabetic retinopathy group (76 cases,32 male,44 female,average age 62.97 ± 12.46) and non-diabetic retinopathy group (92 cases,46male,46 female,average age 58.14 ± 13.71),and 65 cases of healthy subjects (46male,46 female,average age 60.24 ± 10.85) were enrolled as normal control group as well.Hcy and HbA1c were detected in all groups.Multiple independent samples nonparametric tests were used to compare the difference between groups.The concentrations of Hcy and HbA1c between DR group and NDR group were analysised by ROC curve analysis.Result The Hcy levels in normal control group,NDR group and DR group were 7.00 (4.55-10.50) μmol/L,12.45 (10.30-15.10) μmol/L,20.10 (16.63-25.68) μmol/L respectively.The HbA1c levels in normal control group,NDR group and DR group were 5.30 (5.00-5.70) ％,6.80 (5.83-8.03) ％,7.25 (6.50-8.60) ％ respectively.There was no significant difference in Hay and HbA1c levels between groups (P ＜ 0.01);Compared with non-diabetic retinopathy group,levels of Hcy,HbA1c in diabetic retinopathy group were obviously higher;The optimal diagnostic cut-off values of Hcy was 14.35 μmol/L,at this cut-off value,the sensitivity was 96.05％,specificity was 68.48％;The optimal diagnostic cut-off values of HbA1c was 6.05％,at this cut-off value,the sensitivity was 98.68％,specificity was31.25％.Conclusion Patients with Diabetic retinopathy might have hyperhomocystinemia,and detection of Hcy may contribute to the early diagnosis of diabetic retinopathy.

Adult ; Echocardiography ; Heart Atria ; pathology ; Heart Diseases ; complications ; diagnostic imaging ; Humans ; Male ; Pulmonary Embolism ; complications ; diagnostic imaging ; Thrombosis ; complications ; diagnostic imaging

OBJECTIVE:To establish a method for the determination of related substances in bifonazole raw material. METH-ODS:HPLC method was performed on the column of Kromasil C18 with mobile phase of methanol-0.02 mol/L phosphoric acid(ad-justed pH to 7.5 with triethylamine)(70:30,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 258 nm,volume injection was 20 μl,and the column temperature was 40 ℃. RESULTS:The linear range was 0.05-0.25 μg/ml for bifonazole(r=0.9996), 0.05-0.25 μg/ml for impurity A(bifonol)(r=0.9997)and 0.05-0.25 μg/ml for impurity B(4-C isomer)(r=0.9995);the detec-tion limits were 8.2 ng/ml,7.5 ng/ml and 8.4 ng/ml,and the quantification limits were 27.1 ng/ml,24.7 ng/ml and 27.8 ng/ml,re-spectively;RSDs of precision and reproducibility tests were lower than 1%;recovery of impurity B was 95.13%-101.29%(RSD=1.89%,n=9);both impurity A and impurity B were were detected in the 3 batches of samples. CONCLUSIONS:The method is accurate,sensitive and reproducible,and can be used for the determination of the related substances in bifonazole raw material.

OBJECTIVE To establish a rapid detecting method for Clostridium perfringens on traumatic tissues by type-specific multiplex PCR.METHODS Established a simple and rapid method(TLS method) for purifying the DNA of genomes and plasmids in standard strains of C.perfringens wild strains on surface of open traumatic tissues and detected DNAs by type-specific multiplex PCR.RESULTS All types of C.perfringens could be detected by type-specific multiplex PCR.The sensitivity by PCR for type A of C.perfringens arrived at 7.5?103/ml and a whole run could finish within 5 h;the results by PCR entirely corresponded with those by cultureing.CONCLUSIONS New methods for purifying DNA of genomes and plasmids of C.perfringens are simple and rapid;there are high specificity and sensitivity for detecting DNA by multiplex PCR within short time,which can be practiced in clinical laboratory.