Objective; To clone, sequence and analyze the coding sequences of the Mr 15000 and Mr 9000 active segments of natural granulysin derived from human CTLs activated by allogenic antigen ,in order to establish the basis for further purifying and investigating the immune - impairing mechanism of granulysin. Methods; The coding sequences of the Mr 15000 and Mr 9000 granulysin gene were amplified from the total RNA of activated CTLs of healthy person after reverse transcription by Nested - PCR, inserted into pET32a ( + ) vectors and then transformed into E. Coli TOP10, respectively. The recons were identified by PCR, endonuclease digestion and sequencing. Results: The whole coding sequences of the Mr 15000 and Mr 9000 active segments were successfully cloned as expected. The accurate pET32a - Mr 15000 and pET32a - Mr 9000 recons were obtained through Colony - PCR, endonuclease digestion and sequencing. There lied polymorphism on the 119 th amino acid of the product encoded by GLS gene. Conclusion; The coding sequences of the Mr 15000 and Mr 9000 active segments of human granulysin can be obtained and cloned by the methods mentioned above and can be used for subsequent research.

0.05).It could also decrease the times of wet manure induced by folia sennae,while it was of no effect on diarrhea induced by castor oil. CONCLUSION: Volatile oil from A.longiligulare T.L.Wu has anti-inflammation,analgesic and anti-diarrhea effect related to the cure for ulcerative colitis.

AIM: To investigate antioxidative and antinitrosative effects of volatile oil from A.longiligulare T.L.Wu on dextran sulfate sodium(DSS)-induced ulcerative colitis mice. METHODS: Balb/c mice were fed with 4% DSS solution for 7 d to induce ulcerative colitis.Using biochemical method,the activity of antioxidative enzyme SOD,MDA and NO were determined in normal,model,SASP and three mouse's groups with low,moderate and high volatile oil from A.longiligulare T.L.Wu respectively.At the same time,the activity of iNOS was also measured by immunohistopathology. RESULTS: The concentration of MDA and NO were reduced and SOD increased significantly in high-and moderate-volatile oil groups compared with those in the model group.The activity of iNOS was reduced significantly in high-and moderate-volatile oil groups compared with those in the model group.The results demonstrated that the expression of iNOS was significantly inhibited in DSSinduced ulcerative colitis mice after being treated with high or moderate-dosage volatile oil. CONCLUSION: Volatile oil from A.longiligulare T.L.Wu has antioxidative and antinitrosative effects which may be one of the mechanism for treating UC.

OBJECTIVESTo investigate the results of two-part surgical neck fractures of proximal humerus treated with locking intramedullary nail for proximal humerus, and to discuss the indication, surgical technique and the effectiveness.

METHODSThe data of the 22 patients with two-part surgical neck fractures fixed with proximal humeral nail was retrospectively reviewed. The mean age was 57 years. X-ray film and visual analogue scale (VAS), American shoulder and elbow surgeon (ASES) score, Constant-Murley score, University of California, Los Angeles (UCLA) scoring system and simple shoulder test (SST) questionnaire were used to evaluate the patients' shoulder function recovery at final follow-up.

RESULTAll the patients were followed up with an average time of 13 months. All fractures united within 8 weeks postoperatively. No infection, avascular necrosis or complications which concerning with hardware was found during follow-up. The average active forward flexion was 147.8 degrees , the average external rotation was 45.5 degrees and the average internal rotation was T10. The mean VAS score was 1.5. The mean ASES score, Constant-Murley score, UCLA score and SST score were 81.2, 85.4, 29.9 and 9.5 respectively. Eighteen patients were rated as "excellent" or "good", while 4 others were rated "poor". All patients were satisfied with their results.

CONCLUSIONWith good indication control, meticulous intraoperative management and strict postoperative rehabilitation, close reduction and internal fixation with proximal humeral nail can be an effective method for the treatment of two-part surgical neck fracture.

Adult ; Aged ; Bone Nails ; Female ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; instrumentation ; methods ; Humans ; Humerus ; surgery ; Internal Fixators ; Male ; Middle Aged ; Retrospective Studies ; Shoulder Fractures ; surgery ; Treatment Outcome

OBJECTIVETo evaluate the indications and clinical results of displaced proximal humeral fractures treated by locking proximal humeral plate (LPHP).

METHODSFrom September 2004 to March 2006, 75 patients which were treated by open reduction and internal fixation with LPHP were available to follow-up, with an average time of 17 months (12 - 30 months). There were 60 fresh fractures and 15 delayed fractures. The range of motion, muscle strength, visual analogue scale (VAS) for pain, American shoulder and elbow surgeon (ASES) score, Constant-Murley, University of California, Los Angeles (UCLA) scoring system and simple shoulder test (SST) for function evaluation were all recorded.

RESULTSThe average forward flexion was (147.3 +/- 17.7) degrees , external rotation was (30.5 +/- 16.2) degrees and internal rotation was to T9. The mean ASES was 86.7 +/- 12.7, Constant-Murley was 87.4 +/- 10.5, UCLA was 30.1 +/- 4.2. The total good or excellent rate was 89.3%. Compare with fresh fractures, the delayed group showed significant less forward flexion and ASES (P = 0.021 and 0.036 respectively). In 9 patients with late complications, there were significant differences regarding ASES, UCLA scoring system and Constant-Murley compared with patients without any complication.

CONCLUSIONWith strict indication control and appropriate surgical technique, satisfactory results can be expected for the displaced proximal humeral fractures treated with locking proximal humeral plates.

Adult ; Aged ; Aged, 80 and over ; Bone Plates ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Shoulder Fractures ; surgery ; Treatment Outcome

Objective To observe the role of rosiglitazone in unclipped kidneys of two-kidney- one-clip hypertensive rats and examine its relationship to angiotensinⅡreceptors.Methods Two- kidney-one-clip hypertensive rats were divided randomly into 4 groups as follows:positive control group (CONT),traditional antihypertensive drugs group (TAHD,reserpine 50?g?kg~(-1)?d~(-1), dihydralazine 6.25 mg?kg~(-1)?d~(-1) and hydrochlorothiazide 6.25 mg?kg~(-1)?d~(-1),regular-dose rosiglitazone group (RRGL,rosiglitazone 5 mg?kg~(-1)?d~(-1)),and high-dose rosiglitazone group (HRGL, rosiglitazone 20 mg?kg~(-1)?d~(-1)).Sham operation rats were as negative controls.Each group had 8 rats. Animals were monitored and sacrificed at 10th week.Results Blood systolic pressure in TAHD group and HRGL group was significantly lower than that in CONT group [TAHD(137?27 ) mm Hg and HRGL (143?16) mm Hg vs CONT (191?25 ) mm Hg,P＜0.05],but no significant difference between the former two groups was found.Nor did the blood systolic pressure between RRGL group [(176?18) mm Hg] and CONT group.At 10th week,rats in SHAM group and treated groups had lower urinary urinary protein excretion rate,glomerular injury score and wall-to-lumen ratio of arteriole than those in CONT group [vs CONT urinary protein excretion rate (44.60?17.40) mg/24 h,P＜0.05; vs CONT glomerular injury score 60.85?33.05,P＜0.05;vs CONT wall-to-lumen ratio of arteriole 2.33?1.01,P＜0.01,except TAHD group].Though with the similar level of blood pressure,blood glucose and lipid,HRGL,compared with TAHD group showed lower urinary protein excretion rate [HRGL (16.78?3.50) mg/24 h vs TAHD (27.94?12.79) mg/24 h,P＜0.05],decreased glomerular injury score (HRGL 18.04?7.76 vs TAHD 27.92?6.39,P＜0.05) and wall-to-lumen ratio of arteriole (HRGL 1.75?0.38 vs TAHD 2.16?0.90,P＜0.05) in the cortexes of unclipped right kidneys.The expression of type 1 angiotensinⅡreceptor (AT1R) mRNA was no difference in HRGL group and TAHD group,but the expression of type 2 angiotensinⅡreceptor (AT2R) mRNA was more intensive in HRGL group.Conclusion Rosiglitazone can protect the kidneys from hypertensive injury,especially in high dose.The beneficial effects seem incompletely dependent on the metabolism modulating and reduction of blood pressure,but in relationship to the upregulation of AT2R mRNA.

Objective To construct eukaryotic expression recombinant plasmid containing human granulysin(GLS) and investigate the effect of GLS expression in macrophage RAW264.7 cells on the bactericidal activity against intracellular Mycobacterium tuberculosis.Methods GLS gene was amplified by nested-polymerase chain reaction(PCR) from human cytotoxicity T lymphocyte(CTL) activated by allogenic antigen,and inserted into pBudCE4.1 vector to construct recombinant plasmid.Subsequently,the plasmid was transfccted into RAW264.7 cells which were infected with Mycobacterium tuberculosis H37Rv.The expression of GLS was detected by nested-PCR and immunocytochemistry method.The RAW264.7 cells were lysed after transfected for 96 h,then acidfast stained,cultivated and colony count were done to determine the intraeellular bactericidal activity of GLS.The data were analyzed by t or t' test.Results The pBudCE4.1/GLS eukaryotic expression recombinant plasmid was successfully constructed.The transcriptional and translational expressions of target gene GLS were detected in RAW264.7 cells which were infected with Mycobacterium tuberculosis H37Rv.The bacterial load in macrophages of phorbol myristate acetate(PMA)+pBudCE4.1/GLS group,PMA+pBudCEA.1 group and non-activated group were 1.44±1.25,3.16±0.20 and 3.59±0.21,respectively.The differences between groups were all significant (t=2.403,t=2.854,both P＜0.05).Conclusion Eukaryotic expression recombinant plasmid carrying human GLS gene expressed in macrophages has strong bactericidal activity against intracellular mycobacteria,which provide information for the further study on therapeutic vaccine against Mycobacterium tuberculosis.

Objective: To investigate the alteration of bcl- 2 gene family in the rat brain and the molecular mechanism of neuronal apoptosis following traumatic brain injury. Methods: Male Sprague -Dawley rats were subjected to lateral fluid percussion brain injury(FPI) of mo derate severity. Bcl-2, Bcl-x and Bax protein expression was detected by immun ohistochemistry. Results: (1) The immunoreactivity of Bcl-2 and Bcl-x protein decreased in the hippocampus ipsilateral impact site as early as 6 h post-injury, and this was the main cause of down-regulation of the ratio of Bcl-2+Bcl-x to Bax. (2) During 1-3 d after injury, the Bax protein express i on increased significantly, while the Bcl-2 and Bcl-x protein expression decre ased relatively slow. The decreased ratio of Bcl-2+Bcl-x to Bax was mainly due to the Bax up-regulation. Conclusion: The bcl-2 gene family is involved in neuronal apoptosis after FBI, and the protein expression alteration of the family members leads the neuronal cell to apoptosis.

AIM: To study the effect of extract from Pongamia pinnata roots on anti-inflammation and analgesia and acute toxicity. METHODS: The models of mice ear edema induced by xylene and Cotton pellet granuloma in rats to observe the anti-inflammation effect of PRE via oral administration. The effect of PRE on analgesia was tested by measuring the latent period licking hind foot with the hot plate method and counting body twisting induced by acetic acid in mice. The acute toxicity of PRE was measured by the method of Bliss. RESULTS: PRE could significantly inhibit the ear edema caused by xylene in mice, granuloma hyperplasia caused by cotton in rats. It could significantly prolong the pain threshold on hot-plate in mice, reduce the writhing times in mice. The LD50 of PRE was 6. 371 8 g/kg, its 95% confident limit was 5. 408 4-7. 723 2 g/kg. CONCLUSION: PRE has obvious effect on anti-inflammation and analgesia and the lower acute toxicity.

Background The aspheric intraocular lenses(IOLs)can reduce ocular spherical aberration to some degree.However,the clinical effect depends more on the IOL proper alignment.It becomes more important to study the IOL position in eye,Objective This study was to analyze the position alteration of IOL after phacoemulsification combined with implantation of one-piece soft IOLs.Methods In this prospective control study,80 eyes of 40 patients with age-related cataract were enrolled.The phacoemulsification with IOL implantation was performed in all the eyes.Decentration and tilt of IOL in the nasal superior,superior temporal,inferior temporal and nasal inferior quadrants(the intersection point of the system optical axis and the IOL maximum cross plane were regarded as the ordinate origin)were measured by rotating Scheimpflug camera(Pentacam Oculus)in 3 months postoperatively under the mydriasis condition.Written informed consent was obtained from each subject prior to this trial.Results In the right eye group,the IOL decentered toward temporal in 26 eyes(65％)and infratemporal in 16 eyes(40％).IOLs tilted temporally in the horizontal plane in 37 eyes(92.5％)and tilted inferiorly in the vertical plane in 34 eyes(85.0％).In the left eye group,IOLs decentered temporally 33 eyes(82.5％)and 20 IOLs (50％)infratemporally,IOLs tilted temporally in the horizontal plane in 37 eye(92.5％)and 36 IOLs(90％)tilted inferiorly in the vertical plane.There was no statistical difference for the intercomparsion of horizontal/vertical decentration in various quadrant in the right eye(F =0.221,0.792,P＞0.05).The obvious elevated horizontal decentration was found in the supertemporal and infratemporal quadrants compared with supernasal quadrant in the left eyes but there was no significant difference in the vertical decentration among 3 quadrants(F=0.576,P＞0.05).Decentrations were positively correlated with the tilt in both horizontal and vertical plane(right eye horizontal plane:r=0.374,P=0.002;right eye vertical plane:r=0.402,P=0.001 ;left eye horizontal plane:r=0.377,P=0.002；left eye vertical plane:r=0.347,P=0.002).Conclusions The one-piece soft IOLs(Adapt AO)decenter toward temporal mostly in 3 months after surgery,especially infratemporally in the eye.And the optical axis of the IOL tilt toward infratemperol mostly in both right and left eyes.The decentration and tilt are consisted in the corresponding direction between the right and left eyes.The position of the IOLs showed mirror symmetry between right and left eyes.The IOLs decentration show the positively correlation to tilt whatever in horizontal and vertical plane.