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.

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.

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

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.

Transcriptions are regulated by transcription factors. Natural transcription factors usually consist of at least two functional domains: a DNA-binding domain and an effector domain. According to this, novel artificial transcription factors are designed to up or down regulate transcription and expression of a target gene. The Cys2-His2 zinc finger domain is a DNA-binding module that has been widely used as the DNA-binding domain in artificial transcription factors. Each zinc finger domain, which comprises about 30 amino acids that adopt a compact structure by chelating a zinc ion, typically functions by binding 3 base pairs of DNA sequence. Several zinc fingers linked together would bind proportionally longer DNA sequences. According to the "bipartite complementary" library strategy, a pair of zinc finger phage display libraries were constructed. After construction of the libraries, a 9bp sequence (5'-GCAGAGGCC-3') on the promoter of SV40 was chosen as a target for next step. After parallel selection, PCR amplification, desired fragments recovery, re-ligation, and additional rounds of selection, phage enzyme-linked ELISA experiments were performed to identify specific binding clones displaying the zinc fingers with predetermined sequence-specificity to our target sequence. Then two clones with strong ELISA signals were chosen to be tested for binding both to its full target site (5'-GCAGAGGCC-3') and to sites containing single transition mutations. The binding specificity of one of the two clones (clone 3) was shown to be fairly good. The three-finger DNA-binding domain targeted to SV40 promoter, that is, zinc finger sequences on clone 3, was fused to KOX1 suppression domain KRAB and cloned into pcDNA3.1 (+) (which expression product was artificial transcription factor). The zinc fingers (which expression product was the DNA-binding domain of artificial transcription factor) and KRAB domain only (which expression product was effector domain of artificial transcription factor) were also cloned separately into the same expression vector. All constructs contained an N-terminal nuclear localization signal. Every of the vectors (including pcDNA3.1 (+) without inserting sequences) were cotransfected with pGL3-Control and pRL-TK and the activity of luciferase was used to indicate the function of product from transfected expression vectors. Our artificial transcription factor was proved to repress the expression of reporter gene efficiently,while with only DNA-binding domain or effector domain the repression was not remarkable. By adding different effector domains and changing the DNA-binding domain, artificial transcription factor would have a wide range of potential applications.
Enzyme-Linked Immunosorbent Assay ; Genes, Synthetic ; genetics ; physiology ; Models, Theoretical ; Peptide Library ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics ; Transcription Factors ; chemical synthesis ; chemistry ; metabolism ; Zinc Fingers ; genetics ; physiology

OBJECTIVETo compare the clinical efficacy on upper limb motor impairment in stroke between the interactive dynamic scalp acupuncture therapy and the traditional scalp acupuncture therapy.

METHODSThe randomized controlled trial and MINIMIZE layering randomization software were adopted. Seventy patients of upper limb with III to V grade in Brunnstrom scale after stroke were randomized into an interactive dynamic scalp acupuncture group and a traditional scalp acupuncture group, 35 cases in each one. In the interactive dynamic scalp acupuncture group, the middle 2/5 of Dingnieqianxiexian (anterior oblique line of vertex-temporal), the middle 2/5 of Dingniehouxiexian (posterior oblique line of vertex-temporal) and Dingpangerxian (lateral line 2 of vertex) on the affected side were selected as the stimulation areas. Additionally, the rehabilitation training was applied during scalp acupuncture treatment. In the traditional scalp acupuncture group, the scalp stimulation areas were same as the interactive dynamic scalp acupuncture group. But the rehabilitation training was applied separately. The rehabilitation training was applied in the morning and the scalp acupuncture was done in the afternoon. The results in Fugl-Meyer for the upper limb motor function (U-FMA), the Wolf motor function measure scale (WM- FT) and the modified Barthel index in the two groups were compared between the two groups before treatment and in 1 and 2 months of treatment, respectively.

RESULTSAfter treatment, the U-FMA score, WMFT score and the score of the modified Barthel index were all apparently improved as compared with those before treatment (all P < 0.01). The improvement in the U-FMA score after treatment in the interactive dynamic scalp acupuncture group was better than that in the traditional scalp acupuncture group (P < 0.05). For the patients of IV to V grade in Brunnstrom scale, WMFT score in 2 months of treatment and the score of Barthel index after treatment in the interactive dynamic scalp acupuncture group were improved apparently as compared with those in the traditional scalp acupuncture group (P < 0.05, P < 0.01). But, for the patients of III grade in Brunnstrom scale, WMFT score and the score of Barthel index after treatment in the interactive dynamic scalp acupuncture group were not different significantly as compared with those in the traditional scalp acupuncture group (both P > 0.05).

CONCLUSIONFor the patients of IV to V grade in Brunnstrom scale in stroke, the interactive dynamic scalp acupuncture therapy achieves the superior improvements of the upper limb motor function and the activity of daily life as compared with the traditional scalp acupuncture therapy, and the longer the treatment lasts, the more apparent the improvements are. For the patients of III grade in Brunnstrom scale, the interactive dynamic scalp acupuncture therapy achieves the similar improvement in the upper limb motor impairment as compared with the traditional scalp acupuncture therapy.

Activities of Daily Living ; Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Occupational Therapy ; Scalp ; Stroke ; physiopathology ; therapy ; Stroke Rehabilitation ; Treatment Outcome ; Upper Extremity ; physiopathology

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.