Objective To study the growth inhibitory and apoptosis inducting effect of paclitaxel on human osteoblastic cell line U-2 OS. Methods U-2 OS cells were treated with various concentrations of paclitaxel. Proliferation was determined by cell count in a Neubauer cytometer chamber. Viability was assessed by trypanblau dye exclusion. Paclitaxel induced morphologic alterations were visualized, using light and transmission electron microscopy. The extent of paclitaxel induced apoptosis was determined by flow cytometry and immunohistochemical detection (TdT mediated dUTP nick end labeling technique, TUNEL). Results Paclitaxel had a growth inhibitory and apoptosis inducting effect on U-2 OS cell. The cell treated with paclitaxel initially show G2/M arrest; follow by apoptosis. A characteristic apoptosis change including nuclear disintegration and chromatin agglomerate were displayed. Lots of multinucleate cells appeared, which was not seen on the cell treated with other chemotherapeutic agents such as cisplatin and adriamycin. Also, extensive DNA cleavage was detected by immunohistochemical technique. Conclusion Paclitaxel has an obvious growth inhibitory and apoptosis inducting effect on osteosarcoma cell line by induce a G2/M arrest and inhibit the mitosis. The effect of paclitaxel displays a time dependent and dose dependent manner.

Objective To study the causes of rehematomas after operations of traumatic hematomas of perisylvian area.Methods The causes of 50 cases of rehematoma after operation were analyzed retrospectively.Results The big hematoma in primary contusion and laceration of brain happened in 19 cases(38%),delayed epidural hematoma in opposite side in 15 cases(30%),increased intracerebral hematoma in 9 cases(18%),epidural hematoma in primary area in 3 cases(6%),subdural hematoma caused by postoperative lumbaropuncture in 3 cases(6%),hematoma in encephalonecrosis in 1 case(2%).Conclusion Insuitable operation and hemostasis are the main causes of rehemorrhage,and fracture line in the opposite side,and thrombocytopenia are high risk factors of rehematoma.

Objective To evaluate the safety and effectiveness of balloon kyphoplasty combined with physiotherapy in the treatment of osteoporotic vetcrbral compression fractures (OVCFs). Methods A retrospective review was of 12 OVCF cases ( including 16 fracture vertebrae) treated with balloon kyphoplasty was performed. Each patient had also been treated with anti-osteoporotic medication and the Rehabilitation of Osteoporosis Program-Exercise (ROPE) protocol, and each had received a six-month follow-up visit. The following parameters were recorded before the operation and 1 day and 6 months afterward : fracture recurrence, the severity of pain before and after treatment, and the change of vertebral height. The severity of pain was evaluated by using a visual analogue scale (VAS). Results During the treatment, no complication or adverse event was found. The average VAS values preoperation, postoperation and at the 6-month follow-up were 7.6±1.3, 2.1±1.2 and 2.6±1.4, respectively. The average changes in vertebral height preoperation, postoperation and at the 6-month follow-up were 49.2±18% , 74.3 ±14% and 69.8±16% respectively. All of the measures evaluated improved noticeably after the combined treatment. Conclusion Balloon kyphoplasty combined with anti-osteoporotie medication and physiotherapy showed good effects in the treatment of osteoporotic veterbral compression fractures.

Objective To explore the expression and significance of MMP-1 and TIMP-1 in the nucleus pulposus and anulus fibrosus of intervertebral disc with different lesions.Methods From August 2014 to August 2016,34 cases of lumbar disc herniation (the observation group) and 34 case of surgical removal of intervertebral disc in patients with vertebral trauma caused by sudden trauma (the control group) in our hospital were stuided.The nucleus pulposus and anulus fibrosus of intervertebral disc were separated,and the expression of MMP-1 and TIMP-1 were detected by immunohistochemistry.The relationship between MMP-1 and TIMP-1 expression and intervertebral disc degeneration was analyzed.Results The MMP-1 and TIMP-1 expression of nucleus pulposus in the observation group were significantly higher than those in the control group(P>0.05);and the MMP-1 and TIMP-1 expression of nucleus pulposus decreased successively in prominent type,extrusion type,free type lumbar intervertebral disc degeneration(P<0.05).The MMP-1 and TIMP-1 expression of anulus fibrosus in the observation group were significantly higher than those in the control group(P>0.05);and the MMP-1 and TIMP-1 expression of anulus fibrosus decreased successively in prominent type,extrusion type,free type lumbar intervertebral disc degeneration(P<0.05).Patients with high expression of MMP-1 and TIMP-1 in nucleus pulposus and anulus fibrosus had more risk of degenerative disc disease(P<0.05).As a sensitivity prediction of intervertebral disc degeneration,the sensitivity of MMP-1 was 82.35% while the specificity was 94.11%,and the sensitivity of TIMP-1 was 79.41% while the specificity was 88.24%.The prediction sensitivity of intervertebral disc degeneration was 90.91% and the specificity was 97.14% when combined MMP-1 with TIMP-1.Conclusion The expression of MMP-1 and TIMP-1 in the nucleus pulposus and anulus fibrosus of intervertebral disc degeneration was significantly increased,which is of great clinical significance in the diagnosis of intervertebral disc degeneration.

Object To explore the feasibility of breeding genetic modified (GM) medicine by expressing human cytokine in transgenic Chinese materia medica Methods Human interferon ? gene and RANTES gene available from the amplification in vitro were enzymatically excised, recoveried, and inserted into intermediate vectors The recombinants were identified by double enzyme digestion of EcoRⅠand HindⅢ The plasmids were extracted from Escherichia coli and introduced into A tumefaciens, and the transformants harboring binary vectors were screened by addition of antibiotics of kanamycin (Km) and rifampicin (Rif), and the explants of M charantia and P vulgaris were transformed by co cultivation of leaf disks with A tumefaciens strain Results RT PCR was applied to detect the transient expression of human interferon ? gene and RANTES gene in transformed medicinal herbal calli Conclusion The expression of recombinant human interferon ? gene and RANTES gene in transgenic M charantia and P vulgaris cells was firstly reported, which opens an alternative road to antivirus, especially anti AIDS virus, by using transgenic Chinese materia medica

ObjectiveTo investigate the clinical features and risk factors of patient with Wegener's granulomatosis complicated with pulmonary infection.Methods Patients with Wegener's granulomatosis admitted to our hospital in the past 11 years were retrospectively analyzed.Comparisons between groups were performed by t tests or Fisher test.ResultsPulmonary infection occurred in 27 cases with an incidence rate of 29％.Twenty-six percent of pulmonary infections occurred at the initial diagnosis,and 44％ occurred within 6 months,while 30％ occurred later than 6 months.The clinical manifestations of pulmonary infection were productive cough (89％),hemoptysis (63％),fever and fatigue (56％),chest pain and pactoralgia (33％).The most common causative pathogen were bacteria(59％ ),fungi(37％ ),and tubercle bacillus(37％ ).Sinus infection(P=0.01),hypoproteinemia(P=0.03),hypoimmunoglobulinemia (P=0.007),and methylprednisolone pulse therapy(P=0.002) were the risk factors for pulmonary infection.ConclusionThe occurrence of Wegener's granulomatosis complicated with pulmonary infection is high within 6 months.The most common clinical manifestation is productive cough.The most common causative pathogens are bacteria,tubercle bacillus and fungi.Sinus infection,hypoproteinemia,hypoimmunoglobulinemia,and methylprednisolone pulse therapy are risk factors of pulmonary infection.

AIM: To observe the surgical effects of the taumatic lens subluxation and cataract after manual fragmentation and emulsification of nucleus and foldable intraocular lens implantation.
METHODS: A 3. 0mm tunnel limbus incision was operated through the predicted bulbar conjunctiva and sclera on 26 cases ( 26 eyes ) with taumatic lens subluxation ( suspensory ligament rupture range less than 120 ) and cataract (Ⅰ ~ Ⅲ) . And after the manual fragmentation and emulsification of nucleus, foldable intraocular lens was implantated. Intraocular lens loop was imbedded in the middle of the lens zonular ligament breakup to reset the pouch. The surgical complications and postoperative vision changes were observed.
RESULTS:Three month after operation, 22 eyes had a intraocular lens centric position taking up 85% of the whole. Four eyes had a slightly eccentric position ( 1 ~2mm), taking up 15% of the whole. 21 eyes had their visual acuity 0. 5~0. 8, taking up 81% of the whole. Five eyes of visual acuity was 0. 2~0. 8. Within 24h intraocular pressure of 12 eyes (46%) after operation were elevated, and returned to normal after 2~7d. There was no severe complication during operation and postoperation.
CONCLUSION: The manual fragmentation and emulsification of nucleus and foldable intraocular lens implantation of the traumatic lens subluxation and the cataract through the 3. 0mm corneal sclera limbus tunnel incision is a simple and effective surgery.

Objective To investigate the effects of MG132 on diabetic nephropathy (DN) rats induced with streptozocin. Methods Seventy-two male SD rats were randomly divided into three groups: normal control group (NC, n=24), DN group (n=24) and DN treated with MG132 group (DN+MG132, n=24). At the end of 4, 8 and 12 weeks, 24 hour urinary protein excretion rate (UPER) was detected. Morphology of kidney was examined by special staining of periodic acid-schiff (PAS). Renal 26S proteasome activity was determined by quantifying the hydrolysis of S-LLVY-AMC in a fluorescence reader. Urinary malondialdehyde (MDA) level and renal SOD and GSH-PX activity were detected by commercial kits. Renal SOD, GSH-PX and p47phox mRNA expressions were determined by real-time fluorescence PCR. Renal p47phox protein expression wasdetermined by Western blotting. Results Compared with NC group, the DN group showed a significant increased of UPER at week 4, 8, 12 (all P<0.05), of mesangium proliferation and mesangial matrix expansion at week 12. In DN+MG132 group, UPER was significantly decreased compared with DN group at the end of 4, 8 and 12 weeks (P<0.05, respectively), and the glomeruler pathological alteration induced by diabetes was attenuated. Increased renal 26S proteasome activity in DN rats was significantly inhibited after MC132 administration (P<0.05). Moreover, renal p47phox mRNA expression in DN group was 155%, 149% and 120% more than those in NC group at 3 time points (all P<0.05), and so was the renal p47phox protein expression, 139%, 152% and 186% more (all P<0.05). Urinary MDA levels in DN group were 1.95-, 2.04-and 2.62-folds more than those in NC group (all P<0.05). In addition, compared with NC group at 3 time points, in DN group, renal SOD activity was decreased by 23.09%, 33.59% and 53.31% (all P<0.05); renal GSH-PX activity was decreased by 28.57%, 33.06% and 48.76% (all P< 0.05); renal SOD mRNA was decreased by 38.09%, 61.44% and 76.53% (all P<0.05); renal GSH-PX mRNA group was decreased by 29.16%, 37.26% and 62.40% (all P<0.05). Compared with DN group, renal p47phox mRNA and protein expression, and urinary MDA levels were significantly lower in DN+MG132 group (all P<0.05); renal SOD and GSH-PX activity as well as mRNA expression were significantly increased in DN+MG132 group (all P<0.05). Conclusions MG132 treatment can provide renoprotection for DN rats effectively maybe through enhancing renal anti-oxidative ability.

Objective To investigate the effect and significance of regulating endoplasmic reticulum stress on the expression of histone methyltransferases SET7/9 in the kidneys of db/db mice.Methods Db/db mice were randomly divided into two groups according to random number table method:diabetic nephropathy model group (DN group,n=18) and betaine treatment group (DN+B group,n =18),db/m mice were defined as normal control group (NC group,n =18).At the end of 4,8 and 12 weeks,the expression of GRP78,SET7/9,H3K4me2,and monocyte chemoattractant protein 1 (MCP-1) was determined by real-time fluorescence PCR and Western blotting.24-hour urinary protein excretion rate (UPER) and urine MCP-1 were measured by enzyme linked immunosorbent assay (ELISA).The dynamic changes of blood glucose(BG),serum creatinine (Scr),blood urea nitrogen (BUN) were tested by completely automatic biochemistry analyzer.The morphology of kidney was estimated by special staining of periodic acid-schiff (PAS).Results The levels of BG,BUN,UAER and MCP-1 were significantly higher in DN group than those in NC group (P ＜ 0.05),and were in time-dependent manner.Glomerular basement membrane thickening and mesangial cells proliferation began to emerge in DN group at the end of week 4 and mesangial matrix expansion was more obvious at the end of week 12.The mRNA and protein expression of GRP78 and SET7/9 were elevated significantly in DN group as compared to NC group.The H3K4me2 protein expression level was also increased in time-dependent manner.Compared with the DN group,in DN+B group glomerular lesions attenuated and the GRP78 and SET7/9 expression levels obviously decreased (P ＜ 0.05).Furthermore,the levels of BG,BUN,UPER,MCP-1,H3K4me2 in DN+B group were also reduced (P ＜ 0.05).Conclusion Endoplasmic reticulum stress may be the upstream mechanism of mediating the expression of SET7/9 in the kidneys of DN mice.

Objective To investigate the effect of 4-phenylbutyric acid(4-PBA)on the renal pathogenesis of rats with streptozotocin-induced diabetes and its mechanism. Methods Fifty-four male SD rats were randomly divided into three groups:normal control group(NC group,n=18),diabetic nephropathy group(DN group,n=18),diabetic nephropathy plus 4-PBA treatment group(4-PBA group,n=18).At the end of 4,8 and 12 weeks,index of kidney weight/body weight ratio(KI)were measured and calculated.Serum creatinine (Scr),blood urea nitrogen(BUN),urinary MDA levels,urinary SOD activity,and 24 hour urinary protein excretion ram(UAER)were detected by HITACHI automatically.Morphology of kidney wag examined by special staining of periodic acid-schitt (PAS).The p47phox and nitrotyrosine (NT) expression in kidney were determined by real-time fluorescence PCR and Western blotting. Results Compared with the NC group, the DN group rats showed a significant increase of KI(P＜0.05), UAER(mg/24 h) (4.92±0.70 vs 0.26±0.07, 5.29±0.83 vs 0.28±0.08, 5.54±0.81 vs 0.29±0.04,respectively, P＜0.05]for indicated time, mesangial cells proliferation and mesangial matrix expansion at 12 week. However,4-PBA treatment could significantly inhibit the increase of KI (P＜0.05), decrease UAER (mg/24 h) (3.71±0.37, 3.47±0.36, 3.28±0.40, respectively, P＜0.05]for indicated time, and prevent the glomeruler pathological alteration induced by diabetes. Moreover, the mRNA expression of p47phox in the kidney of DN group was 154.72%, 148.60% and 91.95% more than that of NC group (all P＜0.05) for indicated time. The protein expression of p47phox was 118.00%, 140.10% and 177.82% more than that of NC group (all P＜0.05), and the protein expression of NT was 45.29%,59.13% and 89.28% more than that of NC group (all P＜0.05). In addition, urinary MDA levels in DN group were 2.05-, 2.26- and 2.43- folds of NC group, and urinary SOD activities were decreased by 64.78%, 71.29% and 79.32% of NC group. Compared with the DN group, the mRNA and protein expression of p47phox, and protein expression of NT in 4-PBA group were decreased markedly (all P＜0.05) at the end of 8 and 12 weeks. The urinary MDA level was decreased, and the urinary SOD activity was increased significantly in rats with diabetes after 4-PBA treatment for indicated time (all P＜0.05). Conclusion 4-PBA treatment can significantly inhibit the renal pathogenesis of rats with diabetes through inhibition of oxidative stress.