BACKGROUND: In recent years, foreign scholars have found that apolipoprotein E gene was associated with osteoporosis and its complicated hip fracture. National scholars adopted polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to primarily study the correlation of apolipoprotein E gene polymorphism with bone mineral density (BMD) and osteoporosis.OBJECTIVE: To observe the polymorphism of apolipoprotein E gene, and analyze its correlation with primary osteoporosis and osteoporotic fracture.SETTING: Department of Orthopaedics, Nanchang First Hospital.DESIGN: A clinical controlled observation.PARTICIPANTS: Sixty patients with primary osteoporosis (31 males and 29 females), were enrolled, including 30 cases of hip fracture, who were newly fractured and diagnosed according to radiogram, and 30 cases of simple osteoporosis; Thirty healthy subjects (16 males and 14 females)were also involved.METHODS: Sixty outpatients and inpatients with primary osteoporosis were selected from the Department of Orthopaedics, Nanchang First Hospital from January to December 2002, and they were divided into hip fracture group (n=30) and simple osteoporosis (n=30); Another 30 healthy physical examinees were taken as the healthy control group. All the subjects were fasted for more than 12 hours, and the samples of venous blood were collected on the next morning. DNA was extracted from the separated leucocytes. Gene site DNA amplification was analyzed with polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). The individual DNA templates (provided by Daan Gene Diagnosis Center of Sun Yat-sen University) of identified apolipoprotein E2/2, E3/3 and E4/4 homozygotes were analyzed with PCR-SSCP, and the band showed that E3 was in the front, E2 in the middle, followed by E4. They were in electrophoresis with DNA templates of the samples at the same time, and the apolipoprotein E genotype of the template could be identified by the migration rate of the band.MAIN OUTCOME MEASURES: Apolipoprotein E genotypes were observed.RESULTS: All the enrolled subjects were involved in the analysis of results. ① Six genotypes were detected with PCR-SSCP in the 60 patients with primary osteoporosis and 30 healthy controls, including apolipoprotein E2/2, E3/2, 3/3, E4/2, 4/3, 4/4. ② There were very significant differences in the allele frequency of apolipoprotein E4 among the three groups (χ2=17.520, P ＜ 0.01). The allele frequencies of apolipoprotein E4 in the simple osteoporosis group and osteoporotic fracture group were obviously higher that that in the normal control group (χ2=4.904, 16.681, P ＜ 0.01),it was also markedly higher in the osteoporotic fracture group than in the simple osteoporosis group (χ2=4.658, P ＜ 0.01).CONCLUSION: Apolipoprotein E4 is closely associated with primary osteoporosis and osteoporotic fracture. Apolipoprotein E is a useful marker for primary osteoporosis, especially for osteoporotic fracture.

Objective To investigate the incidence of renal injury caused by strong intensity military training in the navy.Methods One thousand three hundred and twenty servicemen of the navy were enrolled in the present study including 568 recruits and 752 veterans.Urine protein and hemoglobin were determined using the dry chemistry method after a five-kilometer armed field race.Urine samples were collected and centrifuged for erythrocyte count under microscope after training.Retinol binding protein(RBP) and N-acetyl-?-D-glucosaminidase(NAG) were measured by enzyme linked immunosorbent assay(ELISA).Detection of creatine kinase(CK) above 950U/L was assumed to be rhabdomyolysis positive.Results The average incidence of hematuria,hemoglobinuria and proteinuria was 2.1%,7.4% and 44 %,respectively,and the incidence of abnormal RBP and NAG was 57.3% and 57.1%,respectively.Significant difference in each of the indexes existed between the recruits and veterans(P

Aim To investigate the influence of Tanshinone ⅡA on cytokine secreted by kuppfer cells via LPS stimulation,and explore its mechanism of therapying chronic hepatic disease. Methods The liver cell and kupffer cells were isolated and the model of liver cell injuries was made induced by LPS and D-GlaN, and the effects of TanshinoneⅡA was observed on the injuries of liver cell induced by lipopoly-saccharide(LPS) and D-Galactosamine(D-GlaN). Cytokine were released secreted by kupffer cells through LPS stimulation and the contents of TNF-?,IL-6,IL-8 were determined by radio-immunoassay. The hepatocyte morphological character was observed by HE stain,the expressions of TNF-?、CD14、iNOS、eNOS in kupffer cell were explored by immunohistochemical, double-decker chamber was used for observing the damage of hepatocyte caused by cytokine secreted by kupffer via LPS stimulation. Results Tanshinone ⅡA could restore the injury of liver cell induced by D-GlaN and LPS, the levels of ALT、MAO、LDH-L、GSH-S and contents of MDA were significantly reduce.It could inhibit TNF-? IL-8 secreted by kupffer cells through LPS stimulation ,and the expressions of TNF-?、CD14 Inos、eNOS in kupffer cell through LPS stimulation were inhibited. Blocking injuries of the inflammatory cytokine release excessive on liver cell, it hadn′t the protect action on liver cell damage suffer from inflammatory cytokine.Conclusion The mechanism of Tanshinon ⅡA of blocking liver cell injuries induced by D-GlaN and LPS may be correlation with inhibiting the excessive cytokine by kupffer cells

BACKGROUND:Immune rejections after organ transplantation or serious adverse reactions due to immunosuppressive drugs show a lack of effective treatments and poor therapeutic outcomes. Therefore, we try to find an effective immune suprresion method in combination of the latest immunomodulatory achievements. OBJECTIVE:To construct a lentiviral vector overexpressing indoleamine 2,3-dioxygenase (IDO). METHODS:(1) The IDO gene that was successful y contructed was inserted into lentiviral packaging plasmids GV308 to construct GV308-IDO lentivirus packaging plasmids. (2) The 293T cel s with 80%confluence were co-cultured with 5'LTR and 3'LTR, basic elements of lentiviral packaging auxiliary components, including Psi, cPPT, 3FLAG, TetR, IRES, WRPE, TetIIP, Ubiquitin Promoter, SV40 origin and HIV. RESULTS AND CONCLUSION:Western blot assay showed that in 10 g/L agarose gel electrophoresis, there was a target fragment at Mr 48 000. This value was consistent with the size of IDO protein. RT-PCR results showed visible IDO expression in 293T cel s. These findings suggest that IDO fusion gene has been successful y reorganized in the lentiviral packaging plasmids.

OBJECTIVETo investigate the effects of Dragon' s blood extract on proliferation and secret extracellular matrix function of fibroblasts in vitro.

METHODSDragon' s blood was extracted by chloroform, acetoacetic ester, alcohol. Human fibroblast were cultured in vitro in media containing gradient dilutions of Dragon' s blood extracts (0.002, 0.02, 0.2, 2, 20 mg/ml) , which was followed by cell proliferation assessed with MTT assay on 0, 12, 24, 36, 48, 60, 72 h. Under the optimal concentration, the cell growth curves were drawn and the flow cytometry (FCM) was used to determine the changes of cell cycle. On 0, 12, 24, 36, 48, 60, 72 h, the concentration of hyaluronic acid in the supernatant of fibroblast culture was measured by radioimmunoassay.

RESULTS0.2-2 mg/ml Dragon' s blood extracts enhanced the proliferation of fibroblasts in a dose-dependent manner. 2 mg/ml was the optimal dilution of Dragon's blood extract, and it increased the ratio of S cells in cell cycle [(25.80 ± 3.10)%] than control group [(7.50 ± 0.70)%, P < 0.01]. From 12 h to 72 h, in 2 mg/ml Dragon's blood group, concentration of Hyaluronic acid secreted by fibroblasts gradually increased, but were less than control (P < 0.01).

CONCLUSIONSDragon's blood acetoacetic ester extract improved the proliferation of cultured human fibroblasts in vitro, might be beneficial to promote wound healing.

Cell Cycle ; Cell Proliferation ; drug effects ; Culture Media ; chemistry ; Dose-Response Relationship, Drug ; Extracellular Matrix ; Fibroblasts ; cytology ; drug effects ; secretion ; Flow Cytometry ; Humans ; Hyaluronic Acid ; analysis ; secretion ; Plant Extracts ; pharmacology ; Resins, Plant ; Time Factors

BACKGROUND:Dragon’s blood is the main ingredient of traditional medicine prescription for promoting granulation, which has been used in clinical treatment of a variety of refractory wounds and achieved the exact effects. But the Dragon’s blood effect on colagen secretion from normal fibroblasts has not been reported. OBJECTIVE: To investigate the effects of Dragon’s blood extract on the proliferation and secret function of fibroblastsin vitro. METHODS: Dragon’s blood was extracted by extracts chloroform, acetoacetic ester, and alcohol in turn. Normal human fibroblasts were respectively cultured in Dragon’s blood extracts of chloroform, acetoacetic ester, and alcohol, DMEM containing 1% dimethyl sulfoxide, and normal culture medium. Then, the fibroblasts were cultured in vitro in different media containing gradient dilutions of Dragon’s blood extracts (0.002, 0.02, 0.2, 2, 20 g/L), which was folowed by cel proliferation determination assessed with MTT assay. Under the optimal concentration, the cel growth curves were drawn and the flow cytometry was used to determine the changes of cel cycle. The concentration of procolagen type III in the supernatant of the fibroblast culture systems was measured by radioimmunoassay. RESULTS AND CONCLUSION:0.2 g/L-2 g/L dilution of Dragon’s blood extracted by acetoacetic ester enhanced the proliferation of fibroblasts in a dose-dependent manner. The 2 g/L was the optimal dilution of Dragon’s blood extracted by acetoacetic ester, and it increased the ratio of S cels in cel cycle than control group and decreased procolagen type III. These findings indicate that Dragon’s blood acetoacetic ester extract can improve the proliferation of cultured human fibroblastsin vitro, and decrease the secretion of procolagen type III of fibroblasts, and it can be beneficial to improve wound healing and inhibit hypertrophic scar.

Objective To explore the long-term dialysis therapies suitable for Chinese patients by comparison of the long-term clinical effect of daily ambulatory peritoneal dialysis ( DAPD ) and hemodialysis (HD) on patients with end-stage renal failure.Methods Forty-six outpatients and inpatients with end-stage renal failure were enrolled from our hospital and divided into group A (24 patients) and group B (22 patients) randomly.Participants in group A received HD treatmant,including conventional HD,hemodiafiltration (HDF),hemoperfusion (HP),etc.For three times in total 12 - 15 h each week;patients in group B received DAPD with a daily dialysate dose of 6 - 8 L and a replacement in every 3 - 4 h for each bag of peritoneal dialysate,as well as a break overnight.The subsequent long-term monitoring was performed,and the following variables were compared for the patients in these two groups:sufficiency of dialysis (i.e.Kt/V),nutritional status [ including SGA and sALB ),complication control ( including Hb,CO2CP,K+,Ca2+,P-,iPTH and blood pressure (MAP) ],infection incidence,employment rate and annual medical expense.The evaluation on daily activities of living was also performed using Barthel Index (BI) and the presence of depressive dsymptoms was assessed by HAMD.Results Patients in both group A and group B could survive under long-term dialysis,and there was no statistically significant difference in sufficiency of dialysis (Kt/V),nutritional status (SGA and sALB ),complication control ( Hb,CO2 CP,K +,Ca2 +,P -,and MAP) and infection incidence ( P ＞ 0.05 ).One year after dialysis,the urine volume [ (274 ± 102) ml vs.( 205 ± 86 ) ml,P =0.017 ],BI ( 82 ± 13 vs.73 ± 11,P =0.044) and employment rate (40.9％ vs.12.5％,P=0.044) of the patients in group B were higher than those of the patients in group A,but the ultra-filtration volume [ ( 1162 ± 124 ) ml vs.( 1542±136 ) ml,P ＜ 0.001 ],iPTH [ (77.5 ± 12.7 ) ng/L vs.( 104.4±11.3 ) ng/L,P ＜ 0.001 ],HAMD ( 8 ± 3 vs.18 ± 4,P ＜ 0.001 ) and annual medical expense [ (7.67±1.48 ) ng/L vs.( 11.35 ± 2.52 ) ng/L thousand yuan,P ＜ 0.001 ] were lower than those of the patients in group A.Five years after dialysis,B1 ( 80 ± 13vs.71 ± 14,P =0.029 ) and employment rate ( 36.4％ vs.8.3％,P =0.032 ) in group B were greater than those in group A,while iPTH [ (83.8 ± 13.4) ng/L vs.( 123.8 ± 12.4) ng/L,P ＜ 0.001 ],HAMD ( 8±2 vs.19 ± 2,P ＜ 0.001 ),and annual medical expense [ ( 10.32±1.64 ) thousand yuan vs.( 13.47 ±2.38 ) thousand yuan,P ＜ 0.001 ] were lower than those in group A,and there was no statistically significant difference in survival rate (70.8％ vs.86.4％,P=0.289) and dialysis duration [ ( 56.82 ± 6.13 ) mouths vs.(57.35 ± 6.30) months,P =0.774 ] between the two groups ( P ＞ 0.05 ).Conclusion The comparison between the patients treated by long-term DAPD and those treated by conventional HD shows that DAPD has a satisfactory clinical effect in the aspects of sufficiency of dialysis,maintainence of nutritional status,control of complications,regulation of blood pressure and decrease of infection incidence,and it is also superior in reducing depressive symptoms,ensuring better quality of life and mitigating economic burden of patients.

Objective To compare the dose distribution of reserved planned tangential beam IMRT(2-field IMRT) with multifields beams IMRT(6-field IMRT) of the bulk-recurrent chest wall in postmastectomy breast cancer patients.Methods For 8 patients with chest wall in postmastectomy breast cancer bulk-recurrence,2-field IMRT and 6-field IMRT plans were generated on PTV in Pinnacle Planning System.The prescribed dose of PTV was 50 Gy in 25 fractions and GTV was 66-70 Gy which irradiated incrementally by subsequent plan in 8-10 fractions.The conformal index (CI) and homogeneity index (HI) of 95％ of prescribed dose over PTV were compared,while the dose distribution on ipsilateral lung and heart were evaluated.Results The CI of PTV by 6-field IMRT (0.66 ± 0.08) was higher than that of the2-field IMRT (0.53±0.10)(t=7.99,P＜0.05).The HI ofPTV by6-field IMRT (1.36±0.08)waslower than that of 2-field IMRT (2.19 ±0.78) (t =9.04,P ＜0.05).There was no statistical difference in V5,V10,V20 and V35 for ipsilateral lung and in D V35 and D for heart between two plans.Conclusions Compared with 2-field IMRT,6-field IMRT might have a better dose distribution on planning target volume(PTV) for chest wall in postmastectomy breast cancer bulk-recurrence patients,butthere is no significant difference in dose-volume of ipsilateral lung and heart between two plans.

Objective To investigate the diagnosis and the curative effects of interventional therapy by spiral CT in rupture of hepatocellular carcinoma(HCC). Methods 46 patients were checked by spiral CT and abdominal puncture of non-condensable blood. All the patients were treated by the transcatheter arterial embolization (TAE) with iodinated oil and/or gelatin sponge. Results Characteristic findings were detected in 31 patients,of which 7 patients were chiefly detected by spiral CT. The bleeding of ruptured HCCs was controlled effectively in all the cases treated by TAE without serious complications. Conclusion Spiral CT is susceptive to the diagnosis of the ruptured HCC. TAE can be the chiefly treatment in ruptured HCC with fewer serious complications.

ObjectiveTo observe the clinical characteristics and influential factors of pulmonary hypertension (PH) in maintenance hemodialysis (MHD) patients.MethodsOne hundred and two patients with MHD patients whose therapy time were all more than one year were enrolled from June 2008 to May 2009.Pulmonary arterial systolic pressure (PASP) and parameters related with cardiac structure and function were checked out by echocardiography.These patients were divided into two groups according to PASP:PH group[PASP≥35 mm Hg( 1 mm Hg =0.133 kPa)] and non-PH group(PASP ＜ 35 mm Hg).Gender,age,dialysis time,dialysis access and urine volume during dialysis was recorded.The blood biochemistry,parathyroid hormone(PTH),hemoglobin,postdialysis urea nitrogen was recorded too and Kt/V was calculated.ResultsPH was detocted in 26 MHD patients (25.5％,26/102).Right atrium transverse diameter,right atrium long diameter and pulmonary artery internal diameter in PH group [(32.00 ± 8.13),(41.00 ±9.15),(24.38 ± 3.88) mm] were higher than those in non-PH group [(27.44 ± 4.43),(36.45 ±8.88 ),(21.78 ± 2.46) mm] and LVEF in PH group [(55.59 ± 7.88 )％] was lower than that in non-PH group [(59.09 ± 5.69)％] (P ＜ 0.01 or ＜ 0.05).There were significant differences in calcium-phosphorus product,PTH,serum phosphorus and hemoglobin between PH group and non-PH group [(61.36 ± 17.83)mg2/dl2 vs.(53.24 ± 18.28) mg2/dl2,(365.95 ± 178.65) ng/L vs.(287.02 ±209.91 ) ng/L,(2.24 ±0.60)mmol/L vs.( 1.95 ± 0.68) mmol/L,(98.65 ± 25.69) g/L vs.( 114.29 ± 22.31 ) g/L] (P ＜ 0.05 ).Logistic regression found that PTH levels,hemoglobin and calcium-phosphorus product were the major risk factors for PH.ConclusionsMorbidity of PH in MHD patients is obviously high.Secondary hyperparathyroidism,anemia and high calcium-phosphorus product level may take part in the pathogenesis of PH in MHD patients.