Objective:To prepare PEG-SH modified GNPs/miR-29 nanoparticles and to investigate the proliferation and differentiation of neural stem cells induced by PEG-SH modified GNPs/miR-29 nanoparticles.Methods:PEG-SH modified GNPs/miR-29 nanoparticles were developed by oxidation-reduction method and were tested for UV absorption spectrum, particle size distribution and zeta potential of nanoparticles. A total of 15 adult male Sprague Dawley (SD) rats were used to establish spinal cord injury model by modified Allen method. The artificial miR-29 and PEG-SH modified GNPs/miR-29 nanoparticles were implanted into the injury site of spinal cord respectively. The stability of miR-29 expression was analyzed by gel electrophoresis. The neural stem cells were isolated and cultured from 10 SPF grade neonatal rats. It was identified by Nestin, GFAP and NSE antibodies. The activity and proliferation of neural stem cells in synthetic miR-29, PEG-SH GNPs and PEG-SH GNPs/miR-29 nanoparticles group was detected by CCK-8 assay. Neural stem cells were cultured with synthetic miR-29, PEG-SH GNPs and PEG-SH GNPs/miR-29 nanoparticles for 1 week. The density, length and number of neuritis were investigated.Results:The solution of PEG-SH modified GNPs showed a brownish red appearance. The spheres were in uniform distribution under transmission electron microscope. The results of UV absorption spectrum showed a single peak wave. The peak value of UV absorption was near 523 nm. The zeta potential increased gradually with the increased content of PEG-SH. The peak value of zeta potential was 22.5±5.2 mV. With the increase of content of PEG, the particle size of PEG-SH modified GNPs rapidly reached peak value at the early stage and then decreased rapidly to a relatively stable level. The synthetic miR-29 and PEG-SH modified GNPs/miR-29 nanoparticles were implanted into the injury site of spinal cord. At 0-6 h, clear band was observed in the synthetic miR-29 group. However, the band was disappeared rapidly at 12-24 h. In PEG-SH GNPs/miR-29 group, clear band were always observed. The OD values of miR-29 group were 0.34±0.17, 0.78±0.31, 1.28±0.68, 1.64±0.38 at 1, 3, 5 and 7 d after inoculation respectively. There was no significant difference in OD values compared with DMEM group. There was no significant difference in OD values among GNPs, PEG-SH GNPs, PEG-SH GNPs/miR-29 and DMEM group. The density (56.38±3.65 μm 2), length (78.25±3.72 μm) and the number [(356±34.52) /1,000×high power field] of neurites in PEG-SH GNPs/miR-29 group were higher than those in miR-29 group, PEG-SH modified GNPs group and saline group. However, there was no significant difference in the density, length and number of neurite between PEG-SH GNPs/miR-29 and serum group. Conclusion:PEG-SH modified GNPs/miR-29 nanoparticless have good biological properties. It can induce the proliferation and differentiation of neural stem cells with protective effects on miR-29.

OBJECTIVETo determine the enhancement effects of caffeine on chemotherapy of transplanted osteosarcoma in Fischer 344/N rats.

METHODSOsteosarcoma-bearing Fischer 344/N rats were treated with cisplatin 2.5 mg/kg (Group DDP), caffeine 90 mg/kg x 2 d (Group caffeine), and cisplatin 2.5 mg/kg plus caffeine 90 mg/kg x 2 d (Group DDP+caffeine), and the control group was treated with normal saline in the same volume. All drugs were given by intra-peritoneum injection with micro-pump, in the rate of 0.5 ml/h. The tumor volume was measured and evaluated. The tumors were stained in TUNEL, and PCNA was detected with immunohistochemistry. The tumor growth inhibition rate, PCNA index and apoptosis index were calculated, and the survival time were recorded.

RESULTSThe tumor inhibition rate was -0.5219 +/-0.1429 in control group, 0.0362 +/-0.0957 in Group DDP, -0.4193 +/-0.1345 in Group caffeine, and 0.3646 +/-0.1313 in Group DDP+caffeine (P <0.01). PCNA index was 0.4587 +/-0.1312 in control group, 0.1847+/-0.0535 in Group DDP, 0.4381 +/-0.0706 in Group caffeine, and 0.0314 +/-0.0231 in Group DDP+caffeine (P <0.01). Apoptosis index was 0.0008 +/-0.0005 in control group, 0.0077 +/-0.0060 in Group DDP, 0.0011 +/-0.0003 in Group caffeine, and 0.0295 +/-0.0069 in Group DDP+caffeine (P <0.01). And the survival time was (33.63 +/-4.63)d in control group, (52.13 +/-11.74)d in Group DDP, (35.63 +/-5.15)d in Group caffeine, and (55.13 +/-16.23)d in Group DDP+caffeine (P <0.01).

CONCLUSIONCaffeine could enhance the anti-tumor effect of cisplatin in rat osteosarcoma.

Animals ; Antineoplastic Agents ; therapeutic use ; Bone Neoplasms ; drug therapy ; pathology ; Caffeine ; therapeutic use ; Cisplatin ; therapeutic use ; Drug Synergism ; Neoplasm Transplantation ; Osteosarcoma ; drug therapy ; pathology ; Rats ; Rats, Inbred F344

OBJECTIVETo observe the cytotoxic effect of thermo-chemotherapy with cisplatin on osteosarcoma OS-732 cell line and to explore its mechanism.

METHODSThe osteosarcoma OS-732 cell line was treated with different temperature, cisplatin alone and 43 degrees C +cisplatin for 1 h, respectively and the in vitro cytotoxic effect was observed with MTT assay. The cell cycle and the apoptotic rate were analyzed with flow cytometry (FCM); the cellular apoptosis was observed also with electron microscope.

RESULTThe cytotoxicity index increase markedly as the temperature elevated or the concentration of cisplatin increased. While treated with 43 degrees C hyperthermia and cisplatin simultaneously, the cytotoxicity index increased to 72.37%;the cell cycle of the treated OS-732 cells line was changed with marked increase in S phase and decreasing in G(2)/M phase. The apoptotic rate increased markedly with the highest of 56.47%. Electron microscope showed the characteristic apoptotic alterations.

CONCLUSIONHyperthermia with cisplatin enhance cytotoxicity on osteosarcoma OS-732 cell line and the induced cell apoptosis may be one of the mechanisms of enhanced cytotoxicity by thermo-chemotherapy.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bone Neoplasms ; pathology ; therapy ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Combined Modality Therapy ; DNA Damage ; Flow Cytometry ; Humans ; Hyperthermia, Induced ; Osteosarcoma ; pathology ; therapy

PURPOSE: Asthma affects approximately 30 million patients in China; however, tiotropium data for Chinese patients is limited. This study aimed to assess the efficacy and safety of tiotropium in Chinese patients with moderate symptomatic asthma. METHODS: A post hoc subgroup analysis was conducted on 430 Chinese patients pooled from two 24-week, replicate phase 3 trials (NCT01172808 and NCT01172821), in which they received once-daily tiotropium 2.5 µg (Tio R2.5) or 5 µg (Tio R5) (n = 106 or 109, respectively), twice-daily salmeterol 50 µg (Sal 50) (n = 110), or placebo (n = 105), while maintaining inhaled corticosteroids (ICS). The co-primary endpoints assessed in week 24 were forced expiratory volume in 1 second (FEV1) peak0–3h response, trough FEV1 response, and responder rate as assessed using the Asthma Control Questionnaire (ACQ). RESULTS: For both FEV1 peak0–3h responses and trough FEV1 responses, the mean treatment differences were greater for Tio R2.5, Tio R5, and Sal 50 compared with placebo at 0.249 L, 0.234 L, and 0.284 L, and 0.172 L, 0.180 L, and 0.164 L, respectively (P< 0.001). The ACQ responder rate in placebo, Tio R2.5, Tio R5, and Sal 50 was 58.7%, 62.3%, 59.3%, and 69.1%, respectively. Furthermore, 11 (2.6%) of 430 patients had serious adverse events (Tio R5, n = 4; Tio R2.5, n = 1; Sal 50, n = 1; and placebo, n = 5). CONCLUSIONS: Once-daily tiotropium, as add-on to medium-dose ICS, was effective and well tolerated for Chinese patients with moderate symptomatic asthma, consistent with the main analysis.
Adrenal Cortex Hormones ; Adult ; Asian Continental Ancestry Group ; Asthma ; China ; Forced Expiratory Volume ; Humans ; Salmeterol Xinafoate ; Tiotropium Bromide

OBJECTIVESTo summarize the clinical results in the treatment of spinal tuberculosis with debridement, bone grafting and anterior fixation and to evaluate the safety and the value of this procedure.

METHODSFrom June 1997 to May 2001, 18 patients with spinal tuberculosis were treated using anterior debridement, autograft of bone and primary internal instrumentation. They were 8 men and 10 women, aged from 25 to 59 years (mean 41 years). The degree of kyphosis before surgery was 27.0 degrees to 75.5 degrees (mean 47.5 degrees +/- 11.4 degrees ). The involved spines included cervical spine (1 patient), thoracic spine (10), thoracic-lumbar spine (2), and lumbar spine (5). Average 2.8 intervertebral bodies in each patient were afflicted with tuberculosis disease. Spinal fusions were done with iliac bone grafts.

RESULTSAll patients were followed up for an average of 25 months. No deep wound infection and sinus were observed after surgery. The grafted bones were fused in all patients with an average time of 3.6 months. The degree of spine kyphosis correction was 32.7 degrees +/- 8.3 degrees, and 3.2 degrees +/- 2.8 degrees was lost on average in the late stage.

CONCLUSIONAnterior instrumentation for spinal tuberculosis could stabilize the spine, correct kyphosis and fuse the grafted bone.

Adult ; Bone Transplantation ; Debridement ; Female ; Follow-Up Studies ; Humans ; Internal Fixators ; Kyphosis ; microbiology ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; methods ; Treatment Outcome ; Tuberculosis, Spinal ; surgery

Objective: To analyze the effect of a comprehensive intervention on the accuracy of children s body size perception, so as to provide a theoretical basis for child body size perception improvement. Methods: The participants were selected from a cluster randomized controlled trial (September 2018 to June 2019). A total of 1 287 children in 24 primary schools (clusters) equally distributed among three regions (Beijing, Changzhi and Urumqi) were selected, which included 12 intervention schools (648 students) and 12 control schools (639 students). The accuracy of body size perception was measured by Ma figural stimuli. A linear mixed model was employed to analyze the effect of the comprehensive intervention on the accuracy of children s body size perception. Results: At baseline, the accuracy rate of body size perception among children in the intervention group and the control group was 56.6% and 51.5%, respectively. The underestimation rate was 42.0% and 47.7%, and the overestimation rate was 1.4% and 0.8%. After the intervention, compared with the control group, the inaccuracy rate ( OR=0.50, 95%CI=0.37-0.68, P <0.01) and the underestimation rate in the intervention group decreased ( OR=0.37, 95%CI=0.26-0.54, P <0.01). There was no significant difference in the overestimation rate between the two groups( P =0.51). The results of the stratified analysis showed that the intervention could improve the accuracy of children s body size perception, regardless of their gender, nutritional status, region, or whether or not they were only child( P >0.05). Conclusion The inaccuracy rate of children s body size perception, which mainly involved underestimation was high. A comprehensive intervention can effectively reduce body size underestimation and improve the accuracy of children s body size perception.

OBJECTIVETo evaluate the effectiveness and safety of the combination chemotherapy of capecitabine (X) with fractionated administration of cisplatin (C) in Chinese patients with advanced gastric cancer (AGC).

METHODS141 patients with AGC were enrolled between July 2002 and August 2004. All patients had measurable tumor according to the criteria of RECIST, Karnofsky performance status > or = 60, adequate bone marrow, renal and hepatic functions. Prior radiotherapy or adjuvant chemotherapy was not permitted. Patients received oral administration of capecitabine at a dose of 1000 mg/m(2) twice a day on D1-D14, and intravenous infusion of fractionated cisplatin at a dose of 20 mg/m(2)/day on D1-D5. The regimen was repeated every 3 weeks, totally for 6 cycles.

RESULTSOf the 141 evaluable patients, there were 104 men and 37 women, with a median age of 54 years (range, 23 - 80 years). Metastases before chemotherapy were detected in lymph nodes (46.8%), liver (40.4%), lung (5.7%) and other area (10.6%). The median treatment duration was 6 cycles (range, 3 - 6 cycles). The objective response rate (RR) was 36.2% (51/141). The median follow-up period was 17.5 months. The median time to progress (TTP) was 9.0 months, and the median overall survival (OS) was 12.0 months. The most common treatment-related adverse events (grade 3/4) were: hand-foot syndrome (HFS) (2.1%), leucopenia (0.7%), abnormal alanine transaminase elevation (2.8%). There was no treatment-related death.

CONCLUSIONCapecitabine combined with fractionated cisplatin is highly effective and well tolerated as a first-line treatment for advanced gastric cancer, with comparable results to 5-Fu plus cisplatin combination therapy.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Capecitabine ; Cisplatin ; administration & dosage ; adverse effects ; Deoxycytidine ; administration & dosage ; adverse effects ; analogs & derivatives ; Female ; Fluorouracil ; administration & dosage ; adverse effects ; analogs & derivatives ; Follow-Up Studies ; Foot Dermatoses ; chemically induced ; Hand Dermatoses ; chemically induced ; Humans ; Leukopenia ; chemically induced ; Liver Neoplasms ; drug therapy ; secondary ; Lung Neoplasms ; drug therapy ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Remission Induction ; Stomach Neoplasms ; drug therapy ; pathology ; Survival Rate ; Vomiting ; chemically induced ; Young Adult

In this study,we estimated the application value of detecting Mycobacterium tuberculosis (MTB) specific IgG/IgM antibodies for tuberculosis diagnosis with colloidal gold immunochromatography assay (GICA).We collected 332 effective serum samples and their background information,including 260 patients with tuberculosis and 72 healthy individuals.The means of GICA was used to detect MTB specific IgG/IgM antibodies.Results were compared with the clinical diagnosis and the results of bacteriological tests.The SPSS 22.0 software was used to analyze the results,and when P＜0.05 the difference was statistically significant.The sensitivity and specificity of GICA were 41.15％ and 91.67％,and the sensitivity of the bacterial positive and negative patients were 51.38％ and 33.77％,respectively.The positive rate of IgG/IgM antibodies detection with GICA (41.15％) was much higher than that of bacteria with acid-fast stain of sputum smear (18.84％) and sputum bacteria cultivation (36.15 ％) (P ＜ 0.05) respectively.The positive rate of the combination of tuberculosis antibody detection,sputum bacterial culture and sputum smear was 61.54％,higher than the result of single method or combination of two methods.The detection of specific antibodies against MTB in serum with GICA is sensitive,specific,rapid and convenient,which can be used in clinical screening.Meanwhile,there are still certain limitations of this method,and the sensitivity and specificity need to be improved.Therefore,the GICA can be used as an auxiliary diagnosis combined with sputum bacteriology,imaging test and clinical features rather than diagnose tuberculosis alone.

Objective: To investigate the clinical efficacy of long-segment pedicle screw reduction and internal fixation combined with kyphoplasty in the treatment of stage Ⅲ reducible Kummell disease. Methods: The clinical data of 32 patients with stage Ⅲ reducible Kummell disease treated at the Department of Orthopedics, Sir Run Run Shaw Hospital, Zhejiang University College of Medicine from January 2012 to March 2017 were analyzed retrospectively.There were 7 males and 25 females,aged (71.8±6.7)years(range:61 to 86 years).The injured segment was T₁₀ in 1 patient,T₁₁ in 8 patients,T₁₂ in 13 patients,L₁ in 7 patients,L₂ in 2 patients and L₃ in 1 patient.Preoperative American spinal injury association(ASIA) classification of patients all showed grade D.Bone mineral density (BMD),spinal X-ray,CT and MRI were examined before operation.All patients were treated with postural reduction, long-segment pedicle screw reduction and internal fixation combined with kyphoplasty.The operation time,intraoperative blood loss,length of stay and postoperative complications were recorded.The visual analogue scale (VAS) and Oswestry dysfunction index (ODI) as well as the BMD of hip were collected before and after operation.The Cobb angle of involved segment kyphosis and the height of anterior edge of diseased vertebrae were measured before operation,3 days and 12 months after operation.CT-related parameters were measured before and 3 days after operation,including sagittal anterior and posterior diameter of spinal canal,cross-sectional anterior and posterior diameter of spinal canal and cross-sectional spinal canal area.Paired sample t test and repeated measures were used to compare the data before and after operation. Results: All patients received the operation successfully.The operation time was (131.3±16.9) minutes (range:95 to 180 minutes),the blood loss was (82.5±27.1) ml (range:50 to 150 ml),and the length of stay was (8.3±2.4) days (range:5 to 14 days).All patients were followed up for more than 12 months.The VAS decreased gradually at 3 days,3 months,6 months and 12 months after operation,and the differences were statistically significant compared with the VAS before surgery (all P<0.01).ODI at 3,6 and 12 months after surgery was significantly improved compared with that before surgery(All P<0.01).The CT-related parameters at 3 days after operation were significantly higher than those before operation (All P<0.05).At 12 months after surgery,the Cobb angle decreased from (35.2±7.6) ° preoperatively to (4.3±1.7) ° (t=22.630,P<0.01),the height of anterior edge of diseased vertebrae increased from (4.3±1.0) mm preoperatively to (16.9±2.5) mm(t=-25.845,P<0.01),the bone mineral density of hip increased from -(2.2±0.6) preoperatively to -(2.8±0.6)(t=-0.040,P<0.01).Up to the last follow-up,2 patients had distal pedicle screw loosening, 1 patient had proximal junctional kyphosis,and there was no new vertebral fracture. Conclusions: Based on postural reduction,long-segment pedicle screw reduction and internal fixation combined with kyphoplasty is a safe and effective treatment method for stage Ⅲ reducible Kummell disease,which can reconstruct the stability of the diseased vertebrae.Postoperative standard anti-osteoporosis treatment is the basis to ensure the efficacy.
Aged ; Cross-Sectional Studies ; Female ; Fracture Fixation, Internal ; Humans ; Kyphoplasty ; Lumbar Vertebrae/surgery* ; Male ; Pedicle Screws ; Retrospective Studies ; Spinal Fractures/surgery* ; Thoracic Vertebrae/surgery* ; Treatment Outcome

Objective: To study the effects of dihydromyricetin (DMY) on the proliferation, apoptosis and epithelial mesenchymal transition (EMT) of esophageal squamous cell carcinoma (ESCC) cell KYSE150 and KYSE410. Methods: KYSE150 and KYSE410 cells were treated with different concentrations of DMY (0, 25, 50, 100, 150, 200 μmol/L) for 24 hours. The median inhibition concentration (IC₅₀) values of KYSE150 and KYSE410 were detected by cell counting kit-8 (CCK-8) method. Then 0.5‰ dimethyl sulfoxide (DMSO) was used as control group, dihydromyricetin (DMY), dihydromyricetin and transforming growth factor-β1 (DMY+ TGF-β1), transforming growth factor-β1 (TGF-β1) were used as experimental group. Cell proliferation and apoptosis rates were measured by clonal formation and flow cytometry. Transwell invasion and wound healing assay were used to detect cell invasion and migration. The protein expression levels of Caspase-3, Caspase-9, Bcl-2, Bax, Smad2/3, phosphorylation-Smad2/3 (p-Smad2/3) and Vimentin were detected by western blot. Results: The IC₅₀ values of DMY on KYSE410 and KYSE150 cells were 100.51 and 101.27 μmol/L. The clone formation numbers of KYSE150 and KYSE410 in DMY group [(0.53±0.03) and (0.31±0.03)] were lower than those in DMSO group [(1.00±0.10) and (1.00±0.05), P<0.05]. The apoptosis rates of KYSE150 and KYSE410 cells in DMY group [(1.84±0.22)% and (2.80±0.07)%] were higher than those in DMSO group [(1.00±0.18)% and (1.00±0.07)%, P<0.05]. The invasion numbers of KYSE150 and KYSE410 cells in DMY group [(0.42±0.03) and (0.29±0.05)] were lower than those in DMSO group [(1.00±0.08) and (1.00±0.05), P<0.05]. The migration rates of KYSE150 and KYSE410 cells in DMY group [(0.65±0.14)% and (0.40±0.17)%] were lower than those in DMSO group [(1.00±0.10)% and (1.00±0.08)%, P<0.05]. The clone formation numbers of KYSE150 and KYSE410 in TGF-β1 group [(1.01±0.08) and (0.99±0.25)] were higher than those in DMY+ TGF-β1 group [(0.73±0.10) and (0.58±0.05), P<0.05]. The apoptosis rates of KYSE150 and KYSE410 cells in TGF-β1 group [(0.81±0.14)% and (1.18±0.10)%] were lower than those in DMY+ TGF-β1 group [(1.38±0.22)% and (1.85±0.04)%, P<0.05]. The invasion numbers of KYSE150 and KYSE410 cells in TGF-β1 group [(1.19±0.11) and (1.39±0.11)] were higher than those in DMY+ TGF-β1 group [(0.93±0.09) and (0.93±0.05), P<0.05]. The migration rates of KYSE150 and KYSE410 cells in TGF-β1 group [(1.87±0.19)% and (1.32±0.04)%] were higher than those in DMY+ TGF-β1 group [(0.86±0.16)% and (0.77±0.12)%, P<0.05]. The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in DMY group were higher than those in DMSO group, while the protein expression level of Bcl-2 was lower than that in DMSO group (P<0.05). The protein expression levels of p-Smad2/3, Smad2/3 and Vimentin in KYSE150 and KYSE410 cells in DMY group were lower than those in DMSO group (P<0.05). The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in TGF-β1 group were lower than those in DMY+ TGF-β1 group, and the protein expression level of Bcl-2 was higher than that in DMY+ TGF-β1 group (P<0.05). The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in DMY+ TGF-β1 group were lower than those in DMY group, and the protein expression level of Bcl-2 was higher than that in DMY group (P<0.05). The protein expression levels of p-Smad2/3, Smad2/3 and Vimentin in KYSE150 and KYSE410 cells in TGF-β1 group were higher than those in DMY+ TGF-β1 group (P<0.05). Conclusion: DMY can inhibit the proliferation and EMT of ESCC mediated by TGF-β1 and promote cell apoptosis.
Apoptosis ; Caspase 3/metabolism* ; Caspase 9/metabolism* ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Dimethyl Sulfoxide/pharmacology* ; Epithelial-Mesenchymal Transition ; Esophageal Neoplasms/metabolism* ; Esophageal Squamous Cell Carcinoma ; Flavonols ; Humans ; Signal Transduction ; Transforming Growth Factor beta1/pharmacology* ; Vimentin/metabolism* ; bcl-2-Associated X Protein/pharmacology*