BACKGROUND:Retinoid X receptor (RXR) plays a central role in the regulation of intracellular receptor signaling pathways. The activation of RXR has protective effect on H2O2-induced apoptosis of H9c2 ventricular cells in rats. But the protective effect and mechanism of activating RXR in cardiomyocytes against hypoxia/reoxygenation (H/R)-induced oxidative iniury are stillunclear. METHODS:The model of H/R injury was established through hypoxia for 2 hours and reoxygenation for 4 hours in H9c2 cardiomyocytes of rats. 9-cis-retinoic acid (9-cis RA) was obtained as an RXR agonist, and HX531 as an RXR antagonist. Cultured cardiomyocytes were randomly divided into four groups:sham group, H/R group, H/R+9-cis RA -pretreated group (100 nmol/L 9-cis RA), and H/R+9-cis RA+HX531-pretreated group (2.5 μmol/L HX531). The cellviability was measured by MTT, apoptosis rate of cardiomyocytes by flow cytometry analysis, and mitochondrial membrane potential (ΔΨm) by JC-1 fluorescent probe, and protein expressions of Bcl-2, Bax and cleaved caspase-9 with Western blotting. Allmeasurement data were expressed as mean±standard deviation, and analyzed using one-way ANOVA and the Dunnett test. Differences were considered significant whenP was <0.05. RESULTS:Pretreatment with RXR agonist enhanced cellviability, reduced apoptosis ratio, and stabled ΔΨm. Dot blotting experiments showed that under H/R stress conditions, Bcl-2 protein level decreased, while Bax and cleaved caspase-9 were increased. 9-cis RA administration before H/R stress prevented these effects, but the protective effects of activating RXR on cardiomyocytes against H/R induced oxidative injury were abolished when pretreated with RXR pan-antagonist HX531. CONCLUSION:The activation of RXR has protective effects against H/R injury in H9c2 cardiomyocytes of rats through attenuating signaling pathway of mitochondria apoptosis.

Objective To explore the anatomical morphology of femoral trochlear groove and the difference between normal males and females.Methods Eighty healthy volunteers were recruited,including 42 males and 38 females with an average age of 36.2 years (range,21-55 years).All the volunteers without knee unstabilization,pain and wound.CT scan of right femurs were performed and 3-D model were reconstructed.The anatomical parameters of right femoral trochlear groove were measured,which included transepicondylar axis,medial and lateral length of trochlear groove,medial and lateral condylar height,sulcus angle,depth of trochlear groove,transcondylar axis,anterior femoral condylar angle,trochlear groove position,and then compared the morphologic difference of trochlear groove between males and females.Results The average width of transepicondylar axis was 79.21±3.80 mm for males and 70.73±2.91 mm for females (t=-53.40,P=0.00).The minimum sulcus angle was acquired at 45° flexion for males and 42° flexion for females.It was 133.92°±4.76° for males and 132.71°±4.36° for females.The maximum length of transepicondylar axis was acquired at 87° flexion for males and 90° flexion for females.It was 42.36±3.48 mm for males and 39.03 ±3.36 mm for females.The anterior femoral condylar angle decreased with the increasing flexion angle of knee (P＞0.05).The position of the trochlear groove moved laterally with the increasing flexion angle of knee (P＞0.05).Conclusion There is no significant difference between male and female in the geometry of femoral trochlear groove,however there is a significant difference in sizes.Therefore,during design the knee prosthesis,close approximation of size is essential,while gender differences in morphology need not be considered a factor.

Objective To evaluate the efficacy and safety of local infiltration analgesia in the multimodal analgesia protocol.Methods Sixty patients who were scheduled to undergo TKA were randomly divided two groups:local infiltration analgesia (LIA) group (n=30) or the non-local infiltration analgesia (N-LIA) group (n=30).All patients were given Celecoxib 200 mg bid,3 days preoperative,and a single-injection femoral nerve block (SFNB) half an hour before the surgery (ropivacaine 3.3 g/L,30 ml).The LIA group was given local infiltration analgesia with ropivacaine (2.5 g/L,60 ml) and 0.1 mg epinephrine before suture the operative incision.The N-LIA group didn't do the LIA.Both of the two groups didn't use the patient controlled analgesia.The VAS scores,the knee joint range of motion,the muscle strength of quadriceps femoris and the side effects and complications were recorded.Results The VAS scores were lower in LIA group than in the N-LIA group,these scores at 2 h to 48 h after surgery at rest and after 24 h at motion had statistical significance.The range of motion and the muscular strength of quadriceps femoris in the LIA group were better than in the N-LIA group.In the LIA group the use of opioids was less and the side effects were lower.The average length of hospital stay after the operation was shorter in the LIA group than the N-LIA group.Conclusion This multimodal perioperative analgesia protocol that include SFNB and LIA offered improved pain control and minimal side effects to patients undergoing TKA.

OBJECTIVETo report a case with cytomegalovirus retinitis (CMVR) after bone marrow transplantation (BMT). A review of the literature and possible mechanisms were presented.

METHODSCase report and literature review.

RESULTSThe patient received large dose of immunosuppressants after allo-BMT appeared CMV infection. There were bleeding and effusion around the ocular vessels. The patient improved after anti-CMV therapy.

CONCLUSIONPatients who undergone allo-BMT were in immunosuppressed condition, when continuous CMV antigenemia and antigenuremia were detected, associated with characteristic change in retinitis, CMVR could be diagnosed.

Adult ; Bone Marrow Transplantation ; adverse effects ; Cytomegalovirus Retinitis ; drug therapy ; etiology ; Humans ; Male ; Transplantation, Homologous

Objective To establish a method for detecting HBV cccDNA in hepatocytes of chronic hepatitis B patients.Method 21 liver biopsies from the hepatic operation patients in the hospital of jiangsu province,concluding 19 HBV chronic infected patients (10 HBeAg positive patients and 9 HBeAg negative patients) and 4 uninfected patients,HBV DNA(+) serum of hepatitis B patients was thought as rcDNA.To use proteinase K to release HBV cccDNA and genomic DNA,then divide the cell lysis solution into two parts,one for detecting HBV cccDNA,the other for detecting the number of ?-Globin as internal control. Nucleic acid for detecting HBV cccDNA extracted by phenol-chloroform was digested by plasmid-safe ATP dependent DNase which was applied to digest the single strand DNA in rcDNA and ssDNA,then was quantitated by the primers spanning across the nick and SYBR Green Ⅰ dye.The specifity of PCR production was confirmed by the sequence analysis and rcDNA comparison.The significance of the difference of HBV cccDNA level between HBeAg(+) and HBeAg(-) group was analyzed by two group t test.Results The agarose gelelectrophoresis showed the molecular weight of the PCR production was about 350bp.The coincidence rate of PCR production and goal fragement was nearly 99% by sequence analysis.The result of PCR detection of rcDNA group was negative.The positive rate of HBV cccDNA of liver biopsies of HBeAg (+) patients detected by this method was 100%,the level of HBV cccDNA in the liver biopsies of HBeAg (+) patients was higher than HBeAb(+) patients.Conclusions The specificity of the method is proved by agarose electrophoresis,gene sequencing of the PCR product and rcDNA comparison.The quantitative method that use SYBR Green Ⅰ dye and ?-Globin as internal control is more specific,sensitive and economical,and more suitable for clinical purpose.

To evaluate the implications of soluble Fas ligand (sFasL) in acute graft-versus-host disease (aGVHD) and discriminating symptoms of aGVHD from those of infection, plasma levels of sFasL were assessed in 84 plasma samples from 13 patients who had undergone allogeneic BMT by using a sandwich enzyme-linked immunological assay (ELISA). Plasma sFasL levels of the patients before BMT and at different time points during the post-BMT period were measured. Three points were concluded: (1) Plasma sFasL levels were higher in patients with grade II-IV aGVHD than in those with grade 0 or I aGVHD. (2) Plasma sFasL levels in patients with infection were not statistically different from those in patients without infection. (3) In seven patients with grade II-IV aGVHD, the plasma sFasL levels at pre-BMT were much lower than those of the six patients with 0 or I grade aGVHD. sFasL may be useful for the diagnosis of aGVHD and for differentiating aGVHD from other BMT related complications such as infection.

Objective To investigate the phenotypes and genotypes of a hereditary protein C(PC) deficiency pedigree.Methods Imrnunoassay(ELISA)was used for PC antigen and PS antigen; Immunoturbidimetry assay was used for measuring AT antigen;Chromogenic substrate assay was used for measuring the activity of PC,PS and AT in Sysmex 1500 automatic Blood Coagulation Analyzer.Polymerase chain reaction(PCR)for amplification of the fragment of each exon and side sequences of PC gene in 10 members of the 3 generations;Direct DNA sequencing was used to examine the mutation site.Results Among 10 members of the 3 generation pedigree,8 of them had a PC:Ag level of 1.06-1.92 mg/L(normal references 3.00-6.00 rag/L),the activity of PC was between 41% and 67%(normal references 70%- 140%),which was significantly lower than the normal references while the levels of PS:Ag,PS:A,AT:Ag and AT:A were all within normal range.DNA sequencing analysis showed that there was a G to T mutation in exon IX of the PC gene at 12 918 position in 8 members.This mutation resulted in the substitution of terminator TGA for TGG which encoding tryptophan at 372 amino acid.There was a polymorphism in 2 405C/ T,2 418A/G,2 583A/T in the promotor area.Conclusions This pedigree is a type I hereditary protein C deficiency.There is a G12 918T mutation in exon IX of PC gene.This mutation is reported for the first time and there is a polymorphism in 2 405C/T,2 418A/G,2 583A/T in the promotor area.

OBJECTIVETo discuss the clinical effects of open reduction and internal fixation (ORIF) for treatment of patients with Lisfranc injury combined the second metatarsal base comminuted fracture.

METHODSFrom March 2007 to June 2012, 7 patients with Lisfranc injury combined the second metatarsal base comminuted fracture were treated including 5 males and 2 female aged from 22 to 51 years old (means 42 years), 4 of sprain and 3 of traffic injury. According Myerson classification, there was 1 case of type A, 3 of type B and 3 of type C. Kirschner wire was used to fix Lisfranc ligament placing from the medial cuneiform bone to the second metatarsal base during the operation. After the operation American Orthopaedic Foot and Ankle Society (AOFAS) criteria system were applied to evaluate the foot and ankle function. Preoperative and postoperative AP, lateral and oblique X-ray and CT scan were collected for radiographic evaluation.

RESULTSAll patients were followed up from 12 to 20 months (16.8 months in average). According to AOFAS criteria system, 3 cases were excellent result,3 good, 1 fair. All the wounds were primary healing without skin necrosis, infection, Kirschner loose,broken, or other complications.

CONCLUSIONKirschner wire had good clinical efficacy for fixing Lisfranc ligament injury with the second metatarsal base comminuted fracture, and could avoid arthrodesis.

Adult ; Bone Wires ; Female ; Humans ; Male ; Metatarsal Bones ; injuries ; surgery ; Middle Aged ; Tarsal Joints ; injuries ; surgery ; Wound Healing

DNA barcoding is a rapidly developing frontier technology in the world and will be useful in promoting the quality control and standardization of traditional Chinese medicine. Until now, many studies concerning DNA barcoding have focused on leaf samples but rarely on Chinese herbal medicine. There are three issues involved in DNA barcoding for traditional Chinese medicinal materials: (1) the extraction methods for total DNA of the rhizomes of the medicinal materials; (2) intra-specific variation among samples from different places of origin; (3) accuracy and stability of this method. In this study, Gentianae Macrophyllae Radix was used to verify the stability and accuracy of DNA barcoding technology. Five regions (ITS2, psbA-trnH, matK, rbcL, and ITS) were tested for their ability to identify 86 samples of Gentianae Macrophyllae Radix and their adulterants. After improving the DNA extraction method, genomic DNA from all samples was successfully obtained. To evaluate each barcode's utility for species authentication, PCR amplification efficiency, genetic divergence, and species authentication were assessed. Among all tested regions only ITS2 locus showed 100% of PCR amplification and identification efficiencies. Based on the established method, we successfully identified two samples of Gentianae Macrophyllae Radix bought in pharmacy to the original species.

Objective To explore the association of ATP-binding cassette (ABC) efflux pump gene Rv1217c-Rv1218c and the drug resistance of Mycobacterium tuberculosis.Methods A total of 34 Mycobacterium tuberculosis clinical isolates including 24 drug-resistance isolates which were resistant to riffampicin,isoniazid,streptomycin or ethambutol and resistant to at least one second-line antituberculosis drug,and 10 drug-sensitive isolates were involved in this study.The RNA of isolated strains was extracted and then reverse transcribed. Gene expression was performed by real-time polymerase chain reaction (PCR) and data was analyzed by t test and Logistic regression analysis.Results The expressions of Rv1217c in rifampicin-resistant group (2.13 ± 1.89,t =3.44,P＜0.01),isoniazid-resistant group ( 1.84 ± 1.86,t =3.16,P＜ 0.05),streptomycin-resistant group ( 1.86 ±1.96,t=2.78,P＜0.05) and ethambutol-resistant groups (3.36±2.35,t=3.04,P＜0.05) were all higher than sensitive isolates (0.42 ± 0.31).The expressions of Rv1218c in rifampicin-resistant group (2.54±1.84,t=3.82,P＜0.01),isoniazid-resistant group (2.34± 1.84,t=3.72,P＜0.01),streptomycin-resistant group (2.15±1.86,t=3.01,P＜0.01) and ethambutol-resistant groups (3.78± 1.78,t=4.22,P＜0.01 ) were all higher than sensitive isolates (0.65 ± 0.42).The expressions of Rv1217c and Rv1218c in multidrug resistant group were 2.74±2.07 and 3.33± 1.77,respectively,which were both higher than polydrug resistant group (0.79 ± 0.47 and 1.03 ± 0.79,respectively; t =2.91,P＜0.05 ; t =3.84,P＜0.01,respectively).Logistic regression analysis found that high Rv1217c expression was positively correlated with rifampicin resistance and negatively correlated with isoniazid resistance (both P＜ 0.01 ),while high Rv1218c expression was negatively correlated with rifampicin resistance and positively correlated with isoniazid resistance (both P＜0.01 ).High expressions of two genes were both positively correlated with ethambutol resistance and multidrug resistance (both P＜0.01 ) and not statistically correlated with streptomycin resistance (P＞0.05).Conclusions The expressions of ABC efflux pump gene,Rv1217c-Rv1218c,are correlated with multiple drug resistance.The overexpression may contribute to the multidrug resistance of Mycobacterium tuberculosis.