[ ABSTRACT ] AIM: To observe the effect of rapamycin on the apoptosis of mouse astrocytes in vitro.ME-THODS:The astrocytes from C57BL/6J newborn mouse pups were isolated and primarily cultured.The effect of rapamycin on the viability of astrocytes was assessed by MTT assay.The mean fluorescence intensity of SYTOX?Green stain in the astrocytes was detected by fluorescence microplate reader in order to analyze the effects of rapamycin on the cell death in-duced by H2 O2 , ionomycin and/or deferorxamin.DiOC6 (3) staining was used to analyze the mitochondrial membrane po-tential of the astrocytes induced by H2 O2 .Flow cytometry analysis was used to determine the production of ROS in the as-trocytes and mitochondria by staining with H2 DCFDA and MitoSOXTM Red reagent, respectively.RESULTS: Rapamycin at concentration of 0.5 μmol/L protected the astrocytes against cell death induced by H2 O2 or deferoxamine plus ionomy-cin.Rapamycin protected the mitochondrial membrane potential of astrocytes from the injury of H2 O2 .It also reduced the production of ROS in the astrocytes and decreased the level of ROS in the mitochondria.CONCLUSION:Rapamycin re-duces the ROS overload in the mitochondria, keeps mitochondrial membrane potential safety and protects the astrocytes a-gainst apoptosis in vitro.

BACKGROUNDSkin lesions are common manifestations in systemic lupus erythematosus (SLE). It is still unknown what the definite pathogenesis of skin involvement was and whether DNA participated in it. Our study was designed to explore the pathogenetic role and nature of nuclear antigen (DNA) deposited in the skin lesions of patients with SLE.

METHODSThirty skin samples from patients with SLE and 2 normal skin samples were studied. Extracellular DNA was evaluated by indirect immunofluorescence methods. The deposited immune complexes were extracted by cryoprecipitation, and DNA was then isolated with phenol and chloroform. DNA fragment sizes were detected by agarose gel electrophoresis. Finally, 8 different probes were used to analyze the origin of these DNA molecules using Dot hybridization.

RESULTSExtracellular DNA staining was found only in skin lesions, mainly those located in the basement membrane zone, vascular wall, and hair follicle wall. Normal skin and non-lesion SLE skin showed no fluorescence at locations outside the nuclei. There were no differences in the rate and intensity of extracellular DNA staining when comparing active phase to remission phase patients. No relationship was found between extracellular DNA and circulating anti-dsDNA antibodies. Deposited DNA fragments clustered into four bands of somewhat discrete sizes: 20 000 bp, 1300 bp, 800-900 bp, 100-200 bp. Small sized fragments (100-200 bp) were positively correlated with disease activity (P < 0.05, r = 0.407). Dot hybridization showed significant homology of the various extracellular DNA fragments examined with human genomic DNA, but not with DNA from the microorganisms and viruses we examined. There were also homologies between DNA samples from different individuals.

CONCLUSIONSDNA and its immune complexes may contribute to the pathogenesis of skin lesions in SLE. These DNA molecules range in size from 100 bp to 20 kb and may be endogenous in origin.

Antibodies, Antinuclear ; blood ; Antigen-Antibody Complex ; analysis ; DNA ; analysis ; immunology ; Humans ; Lupus Erythematosus, Systemic ; immunology ; Skin ; immunology ; Staining and Labeling

ObjectiveTo explore the correlation between angiotensin converting enzyme (ACE) gene polymorphism and hypertension accompanying atherosclerosis in Li people in Hainan province. MethodsTwo hundred and sixty patients with hypertension accompanying atherosclerosis were selected as hypertension plus atherosclerosis group, while two hundred and seventy-six healthy people were regarded as healthy control group. ACE I/D gene polymorphism was detected by polymerase chain reaction (PCR), and the genotype frequencies and allele frequencies of DD, DI and Ⅱ were investigated. The carotid intimal-medial thickness(IMT)was measured by high-resolution ultrasound technique and mean IMT (MIMT) was calculated. Results(1) In the hypertension plus atherosclerosis group, the genotype frequencies of DD, DI and Ⅱ were 15.0%, 37.3%, 47.7%,respectively, and the allele frequencies of D and I were 33.70% and 66.30%, respectively. In the healthy control group, the genotype frequencies of DD, DI and Ⅱ were 17.8% , 40.6% and 41.7%,respectively, and the allele frequencies of D and I were 38.0% and 62.0%, respectively. There were no significant differences both in the genotype frequencies of DD, DI and Ⅱ, and in allele frequencies of D and I between the two groups (P>0. 05). (2) The age,total serum cholesterol(TC),triglyceride (TG), systolic pressure(SBP), diastolic pressure(DBP), apolipoprotein A(apoA) and apolipoprotein B (apoB) levels were significantly higher in the hypertension plus atherosclerosis group than in the control group(P<0. 05). The high density lipoprotein cholesterol(HDL-C) level was significantly lower in the hypertension plus atherosclerosis group than in the control group(P<0. 05). Logistic regression analysis showed that TG (OR = 2.14), apoA(OR = 360. 39), SBP(OR = 1.21), DBP (OR=1.08) and ACE DD genetype (OR = 0. 30) had correlation with hypertension plus atherosclerosis(all P<0. 05). The MIMT level was significantly higher in ACE DD subset than in DI and Ⅱ subset (P<0.05). ConclusionsThe ACE DD genotype increases the susceptibility of carotid atheroselerosis, which is the risk factor for hypertension accompanying atherosclerosis in Li people in Hainan province. It may be an early predictive factor in atherosclerosis.

Objective To investigate the metabolic alterations in the brain of neonates with HIE and correlate those alterations with clinical grading and prognosis of HIE.Methods Fourty-six eases of full-term neonates diagnosed as HIE clinically were performed MRI and 1~H-MRS,9 healthy neonates without the evidence of asphyxia were studied as controls,1~H-MRS techniques included single voxel proton MRS and two dimensional muhi-voxel chemical shift spectroscopy imaging,point resolved spectroscopy sequence was used for 1~H-MRS.Metabolic changes in the spectroscopy were analyzed in neonates with HIE,and study the relationgship between MRS findings and prognosis.Results(1)The typical 1~H-MRS manifestations of full- term neonates suffering from HIE were as follows:the peaks of Lac were elevated,GLx-? were elevated and NAA were decreased.(2)GLx-?/Cr ratio in control,mild,moderate and severe HIE group was 0.16, 0.21,0.64,and 1.31,respectively.Lac/Cr ratio in control,mild,moderate and severe HIE group was 0.12,0.14,0.19,and 0.26,respectively.There was a significant difference in the ratio of GLx-? and Lac/Cr between HIE group and control group(t=5.01,P

OBJECTIVETo summarize the experiences in the treatment of distal radius fracture by locking pi-shaped plate internal fixation.

METHODSAll the 32 cases (left 11, right 21) of unstable fractures of distal radius treated by locking pi plate fixation. Among them, 11 were male and 21 female with an average age of 36 years (range, from 23 to 67 years). There were 16 cases of type B, 9 type C1 and 7 type C2 according to AO classification. Autogeneic bone grafting was applied in 27 patients. All the 32 cases were followed up. The range of motion of the wrist joint and radiographic parameters including palmar inclination, radial length and ulnar variance were evaluated.

RESULTSAll the patients were followed up for 19 to 28 months postoperatively (mean 25 months). Anatomical reduction was achieved in all the cases. Delayed union or non-union was not observed. According to rating scale of Gartland-Werley, 25 cases got excellent results and 7 good. No complications such as loss of reduction, tendon rupture occurred.

CONCLUSIONLocking pi-shaped plate fixation is a reliable and effective method in the treatment of unstable fracture of distal radius.

Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Radius Fractures ; surgery

OBJECTIVE: To investigate the expression of PCGF1 in rectal adenocarcinoma (READ) and the effect of PCGF1 silencing on proliferation READ cells in vitro. METHODS: The UALCAN and ENCORI online databases were used to analyze the expression level of PCGF1 in READ tissues and normal tissues and its association with the clinicopathological parameters and survival outcomes of patients with READ. The expression levels of PCGF1 were detected in two READ cell lines and a normal rectal epithelial cell line (HcoEpiC cells) using qPCR and Western blotting. Lentiviral vectors were used to construct PCGF1-overexpressing and PCGF1-silenced cell lines, and the proliferative activity of the cells was assessed using CCK-8 assay. The effect of PCGF1 silencing on tumor proliferation in vivo was also evaluated by observing tumorigenicity of the cells in nude mice. RESULTS: PCGF1 was highly expressed in READ tissue (P < 0.001), and its expression levels was correlated with READ stage, differentiation and lymph node metastasis (P < 0.001). A high PCGF1 expression level was associated with a poor survival outcome of READ patients (P < 0.05). In SW837 and SW1463 cells, PCGF1 silencing significantly lowered the proliferative activity of the cells both in vitro (P < 0.05) and in nude mice (P < 0.01). CONCLUSION PCGF1 is highly expressed in READ tissue and may potentially serve as a prognostic biomarker as well as a therapeutic target for READ.
Adenocarcinoma/genetics* ; Animals ; Biomarkers ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Mice ; Mice, Nude ; Polycomb Repressive Complex 1 ; Sincalide

OBJECTIVETo determine the risk factors involved in the typhoon episodes and to put forward and evaluate the intervention measures.

METHODSWe defined a confirmed injury case as: 'a person with fall,scalpel and stab, collision, drowning, injuries and trauma due to flying debris and building collapse, asphyxiation due to entrapment in collapsed buildings by typhoon from 0 am,August 12 to 6 pm, August 14 2004' and a death case as: 'a person with fall, scalpel and stab, collision, drowning, injuries and trauma due to flying debris and building collapse, asphyxiation due to entrapment in collapsed buildings by typhoon from 0 am, August 12 to 12 am, August 18 2004'. We investigated all hospitalized injured cases in ten hospitals and telephoned to those who were not hospitalized and the cases of death. We did case-control study with 1 pair versus 2 cases. 74 cases were selected in ten hospitals. The controls were neighbors of the controls matched by occupation, sex, village, and within 5 years of age without injury in this typhoon. We asked the cases and the controls on their alertness regarding typhoon and what actions taken when typhoon arrived.

RESULTSThere were 392 injury cases in all ten hospitals and 50 death cases. The attack rate of injury was 27.3 per 100 000. The fatal rate was 11.3% with the death rate 3.1 per 100 000. We investigated 209 injury cases and 31 death cases. The number of cases who were injured from 1 to 6 hours before typhoon landing accounted for 64.6% (155) of all cases. The peak of epidemic curve was 4 hours before the landing of typhoon. Data on the analysis of 74 cases and 148 controls revealed that 42% (31) of the cases were outside their homes before and during typhoon compared to 15% (22) of the controls (OR = 3.9, 95% CI: 1.9-7.7). Compared with 20% (30) control persons (OR = 17,95% CI: 4.2-68). 28% (21) cases did not receive the alert of typhoon before it arrived compared with 18% (27) control persons (OR = 3.3, 95% CI:1.3-8.6). 53% (39) of the cases did not pay attention to the alert of typhoon before typhoon arrived.

CONCLUSIONStaying outdoor, not receiving or did not take seriously about the alert of typhoon seemed to be the risk factors of injury by the typhoon episode, suggesting that the government should increase the emergency preparedness and to raise the awareness on risks associated with typhoon.

Cause of Death ; China ; epidemiology ; Cyclonic Storms ; Hospitalization ; Humans ; Risk Factors ; Wounds and Injuries ; epidemiology ; mortality

OBJECTIVETo clone a novel human testis-specific gene TDRG1.

METHODSA new contig of expression sequence tags (ESTs) Hs.180197 was identified from the testis libraries using digital differential display (DDD) to screen the novel human testis-specific gene. To validate the use of bioinformatics approaches in gene discovery, the ESTs Hs.180197, which was predicted to be testis specific, was chosen for further study. Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on different normal tissues to identify the expression of Hs.180197 in human testis. Using bioinformatics methods and IMAGE cloning of this contig, the full-length cDNA sequence of the noval human gene was cloned.

RESULTSThis novel gene was 1197 bp in length, located in chromosome 6p21.1-p21.2. The sequence of the open reading frame was 504-806 bp, as confirmed by RT-PCR and sequencing in human testis. The cDNA encodes a novel protein of 100 amino acids with a theoretical molecular weight of 10 000 and isoelectric point of 6.81. The sequence shares no significant homology with any known protein in the databases. Semi-quantitative RT-PCR analysis of multiple tissues further showed that the novel gene was expressed specifically in adult human testis. Considering a possible relation of this novel gene with the function of human testis, we named this new gene TDRG1 (testis development related gene 1, GenBank accession number: DQ168992).

CONCLUSIONDDD combined with laboratory validation is an efficient method for identifying new human functional genes.

Adult ; Cloning, Molecular ; DNA, Complementary ; genetics ; Databases, Genetic ; Gene Expression Regulation ; Homeodomain Proteins ; genetics ; Humans ; Male ; Molecular Sequence Data ; Organ Specificity ; Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment ; Testis ; metabolism

OBJECTIVETo study the relevant position of the pedicle of C1 to the lateral mass of C(2-4), set up an identification technique for the entry point decision of C1 pedicle screw by using the lateral mass of C(2-4) as anatomic landmarks.

METHODSTwenty cadaver specimens were used to measure the distance from the sagittal midline of spine to the medial border, the midpoint and the lateral border of C1 pedicle or the lateral mass of C2, C3 or C4. The anatomic relation between the measurements data of C1 pedicle and that of the lateral masses of the cervical vertebrae were analyzed, and the technique of C1 pedicle screw fixation was established.

RESULTSThe average medial border of the lateral mass of C2, C3 and C4 was 0.37 mm, 0.27 mm and 0.24 mm lateral to that of C1 pedicle, the average midpoint of the lateral mass of C2, C3 and C4 was 1.18 mm, 1.41 mm and 1.74 mm lateral to that of C1 pedicle, and the average lateral border of the lateral mass of C2, C3 and C4 was 1.96 mm, 2.54 mm and 3.24 mm lateral to that of C1 pedicle, respectively.

CONCLUSIONThere is a steady anatomic location relation between C1 pedicle and the lateral mass of C2, C3 or C4. As well as the lateral mass of C2, the lateral mass of C3 or that of C4 could be convenient anatomic landmarks to determine the location of C1 pedicle and the position of C1 pedicle screw entry point.

Adult ; Cadaver ; Cervical Atlas ; anatomy & histology ; surgery ; Cervical Vertebrae ; anatomy & histology ; surgery ; Female ; Humans ; Male ; Spinal Fusion ; methods