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BACKGROUND: Short-term (1 week) vitreous cryopreservation avoiding the formation of ice crystals has been achieved in preserving tissue-engineered bone composed of adipose-derived stem cells (ADSCs)/demineralized bone matrix (DBM) compound through adjusting particular composition of cryopreservation fluid. However, whether vitreous cryopreservation can be utilized to cryopreserve tissue-engineered bone for long term (12 weeks) and maintain cellular viability and osteogenic function after rewarming remains unclear. OBJECTIVE: To investigate the effects of vitreous cryopreservation on viability and osteogenic function of ADSCs for short-term (1 week) and long-term (12 weeks) cryopreservation. METHODS: ADSCs were isolated from New Zealand rabbits and expended to passage 3. Cells at passage 3 were seeded onto DBM derived from porcine trabecular bone and followed by 1 week osteogenic induction. The tissue-engineered bone was transferred to freezing vials of 2 mL containing vitreous cryopreservation fluid and then directly quenched into liquid nitrogen. The composition of cryopreservation fluid was 30% dimethyl sulfoxide, 70% low glucose-Dulbecco's modified Eagle medium (L-DMEM), 0.8 mol/L trehalose. Following vitrification for 1 week or 12 weeks, the composite of ADSCs/DBM was removed and thawed. After rewarming, ADSCs viability were viewed under confocal laser microscope by staining viable cells with the green fluorescent dye Calcein AM and the red fluorescent dye Propidium iodide at days 1, 3, 7, 11 and 13. The number of cells seeded onto the DBM was assayed by Hochest33258 at days 1, 3, 5, 7, 9, 11 and 13. Meanwhile, alkaline phosphatase (ALP) activity was also assayed by PNP microplate method at days 1, 4, 7, 10, 14 and 21. Osteogenic gene expression including Runx2, OCN, ALP, COL-1 was detected by real- time PCR at days 1, 4, 7, 10, 14 and 21. RESULTS AND CONCLUSION: After cryopreservation of 1 week or 12 weeks, it was found that more red-staining live cells was observed at 1 day post-rewarming by live/dead double staining, and the green-staining live cells increased at 3 days. By Hoechst 33258 assay, it was found that the cell number decreased at 1 and 3 days post-rewarming, compared with pre-cryopreservation. However, a constant increase in the cell number was observed beginning at 3 days, reaching the pre-cryopreservation level at 5 days post-rewarming. By PNP microplate method, it was found that ALP activity reduced at 1 and 4days post-rewarming, but compared with the level of pre-cryopreservation there were no significant difference. However, a constant increase in ALP activity was detected since 4 days. By real-time PCR, osteogenic gene expression including Runx2, OCN, ALP, COL-1 reduced at days 1 and 4, but compared with the level of pre-cryopreservation there was no significant difference. However, a constant increase in the osteogenic gene expression was since 4 days. The cell viability and osteogenic function were observed without significant difference at each time point after rewarming of cells that had undergone vitreous cryopreservation for 1 or 12 weeks. Preliminary findings indicate that vitreous cryopreservation can maintain cellular viability and osteogenic function of tissue-engineered bone. Cryopreservation time (1 and 12 weeks) has no significant effect on the cell viability and osteogenic function of the tissue-engineered bone after rewarming.
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Objective To research the application effect of mobile network PBL in combination with the micro-lesson teaching in surgical practice.Methods The research objects were the clinical medical interns of grade 2012. Students were divided into two groups: the experimental group using mobile network PBL in combination with the micro-lesson teaching method, and the control group using the traditional LBL teaching method. The examination scores, case analysis and discuss performance were compared between the two groups.Results The tests grades, clinical cases analysis, discuss performance of the experimental group were significantly higher than those of the control group (P＜0.05); Active classroom atmosphere, active learning, improve teamwork and interpersonal communication of the experimental group were significantly higher than those of control group, et cetera (P＜0.01).Conclusion The effect of mobile network PBL in combination with the micro-lesson teaching method in surgical practice teaching is very good.
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<p><b>OBJECTIVE</b>To study the effects of erythropoietin (EPO) on serum levels of neuron-specific enolase (NSE) and S-100B in neonates with hypoxic-ischemic encephalopathy (HIE) and the underlying mechanism.</p><p><b>METHODS</b>Forty neonates with HIE were randomly divided into conventional treatment (n=20) and EPO treatment groups (n=20). Twenty healthy full-term neonates born during the same period were randomly selected as the normal control group. The conventional treatment group received conventional treatment, while the EPO treatment group received conventional treatment as well as EPO [200 IU/(kg.d)] which was given by intravenous infusion from the second day after birth. The course of treatment was 7 days. Blood samples of the three groups were collected on the first day after birth (before treatment) and the ninth day after birth (after treatment). Serum levels of NSE and S-100B were measured by double-antibody sandwich ABC-ELISA.</p><p><b>RESULTS</b>Before treatment, the two treatment groups had significantly higher serum NSE and S-100B levels than the normal control group (P<0.01), whereas no significant differences in the levels of NSE and S-100B were observed between the conventional treatment and EPO treatment groups (P>0.05). The serum NSE and S-100B levels on the ninth day after birth were significantly lower than those on the first day after birth in the three groups (P<0.01). After treatment, the serum NSE and S-100B levels were significantly lower in the EPO treatment group than in the conventional treatment group (P<0.05).</p><p><b>CONCLUSIONS</b>Dynamic monitoring of serum NSE and S-100B levels may be helpful for the early diagnosis of HIE and the assessment of brain injury repair in newborns with HIE. EPO may be helpful for the repair of neurons and glial cells.</p>
Subject(s)
Female , Humans , Infant, Newborn , Male , Erythropoietin , Pharmacology , Hypoxia-Ischemia, Brain , Blood , Phosphopyruvate Hydratase , Blood , S100 Calcium Binding Protein beta Subunit , BloodABSTRACT
<p><b>BACKGROUND</b>Diabetes has been associated with increased risk of fracture and impaired fracture healing. The aim of this study was to examine the influence of diabetes on perioperative complications, length of stay and ambulatory ability recovery in individuals with hip fracture, and to determine whether changes could be made to improve treatment outcome.</p><p><b>METHODS</b>The study included 707 hip fracture patients treated at Beijing Jishuitan Hospital between July 2009 and December 2010. The medical history and perioperative complications were compared between non-diabetic and diabetic groups. Length of stay, days awaiting surgery, and days of hospitalization after surgery were also analyzed. Ambulatory ability was compared at 1-year follow-up using the Chi-square test and Fisher's exact test. An independent Student's t-test was used to compare normally distributed continuous data.</p><p><b>RESULTS</b>Patients with diabetes were more likely than non-diabetic patients to develop cardiac perioperative complications (8.9% vs. 3.0%, P = 0.021), urinary tract infections (12.0% vs. 2.8%, P < 0.001), and gastrointestinal symptoms (15.0% vs. 6.8%, P = 0.003). No difference in perioperative complications was observed between the groups. Days awaiting surgery and length of hospital stay were both longer in the diabetic group ((8.0 ± 5.1) vs. (6.2 ± 3.7) days and (16.5 ± 3.8) vs. (13.3 ± 3.8) days, P < 0.001, respectively). Before the occurrence of fracture, patients with diabetes were less likely to be ambulatory outdoors (71.9% vs. 85.9%, P < 0.001) and had more restricted walking ability. After at least 1-year follow-up, similar proportions of patients in the non-diabetic and diabetic groups (16.1% and 15.9%, respectively), who were able to ambulate outdoors before the fracture, became housebound till the final follow-up.</p><p><b>CONCLUSIONS</b>Diabetics are at increased risk of specific complications and have a longer time to surgery and longer in-hospital stay, but generally have similar recovery to non-diabetics thereafter.</p>
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Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Chi-Square Distribution , Diabetes Mellitus , Hip Fractures , Perioperative Period , Treatment OutcomeABSTRACT
Objective To investigate the effect of glycemic control on progress of left ventricular structure and diastolic dysfunction in adolescents with type 1 diabetes mellitus (T1 DM).Methods A total of 36 T1DM adolescent patients(observation group) and 36 age-matched healthy controls (healthy control group),who consulted doctors in Chengdu Women and Children's Central Hospital between Dec.2009 and Dec.2010,were recruited into the study.Patients in the observation group were performed standard treatment for glycemic control.All patients were followed-up for 2 years.At the end of the study,the patients in observation group were divided into 3 subgroups according to the average HbA1 c level:excellent glycemic control group [glycosylated hemoglobin (HbA1 c) ＜ 7.6％],good glycemic control group(7.6％ ≤HbA1c≤9.0％) and bad glycemic control group(HbA1c ＞9.0％).All of the subjects were evaluated by means of echocardiography for assessment of left ventricular structural and functional parameters.Results Left ventricular posterior wall depth (LVPW),left ventricular mass index (LVMI) and isovolumic relaxation time (IVRT) were elevated,while E/A was decreased in the observation group compared with the healthy control group at baseline (all P ＜ 0.05).There was no significant difference between 2 groups in interventricular septum thickness (IVS),left ventricular end-diastolic dimension(LVEDd) and cardiac systolic function(all P ＞ 0.05).Echocardiographic parameters of left ventricular structure and function were unchanged in well-controlled patients.At the end of follow-up,mild-control group and non-control group demonstrated increased IVS,LVPW and LVMI,and exacerbated left ventricular diastolic function in the present of IVRT prolonging and E/A decreasing.Conclusions There is an inclination in T1 DM adolescent patients developing to left ventricular diastolic dysfunction,and improved glycemic control can delay the progress of left ventricnlar hypertrophy,but it can't ameliorate the decreased diastolic dysfunction.Whereas,non-improved glycemic control can accelerate left ventricular remodeling and exacerbate diastolic dysfunction in T1DM adolescent patients.
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<p><b>OBJECTIVE</b>To understand the hepatitis C virus (HCV) genotype distribution in Yantai district of Shandong province, and to explore whether the HCV genotypes was relevant to the injure of liver through the index of liver function.</p><p><b>METHODS</b>Using specific PCR primers to amplify the HCV RNA 5' UTR/NS5B,then PCR products were sequenced by genetic analyzer. The genotypes were identified by alignment to the GenBank reference sequences and construction the phylogenetic tree of 5' UTR.</p><p><b>RESULTS</b>Among 9 unpaid blood donors we detected two kinds of genotypes of 1b and 3a, respectively, 8 cases (88.9%) and 1 case (11.1%). Among 33 cases of hepatitis C patients we detected the 1b, 2a and 6a the three kinds of genotypes, respectively, 22 (66.7%), 10 (30.3%) and 1 (3.03%) cases. Subtype 1b is the advantage of popular genotype in HCV carriers from Yantai district, and the distribution of 1b was no significant difference in the different population (chi2 = 0.796, P = 0.373); The difference of indicator of liver damage in the different genotypes of subjects were significant (P < 0.05), the mean of ALT, AST of 2a-subtype carriers was significantly higher than the 1b-subtype population.</p><p><b>CONCLUSIONS</b>The genetic diversity of HCV in Shandong Yantai district was presented. The main genotypes were 1b-subtype, and 3a and 6a-subtype was detected firstly. The genotype of HCV were relevant to the liver damage indicators, 2a-subtype hepatitis C virus infection in the liver cells may play an important role in the disease process.</p>
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Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , China , Epidemiology , Genotype , Hepacivirus , Classification , Genetics , Hepatitis C , Epidemiology , Virology , PhylogenyABSTRACT
<p><b>OBJECTIVE</b>To compare the difference of effects on hearing injury between punching machine noise and steady state noise.</p><p><b>METHODS</b>The subjects of present study were 100 workers, included 38 workers exposed to punching machine noise from forging shops and 62 workers exposed to steady state noise from drawbench or abrasive dust workshops. The individual noise dosimetries were used to detect noise that workers exposed, and cumulative noise exposure (CNE) was then calculated. On the basis of GBZ 49-2007, the worker hearing was tested, and the results were assessed.</p><p><b>RESULTS</b>There were no differences of CNE between group exposed to punching machine noise and group exposed to steady state noise. The hearing loss rate (55.3%) at high frequency in workers exposed to punching machine noise was significantly higher than that (32.3%) in workers exposed to steady state noise (P < 0.01). CNE and the hearing loss rate at high frequency showed dose-response relationship (P < 0.01).</p><p><b>CONCLUSION</b>when the exposure levels of workers were similar, the hearing injury induced by punching machine noise may be significantly higher than that induced by steady state noise.</p>
Subject(s)
Adult , Humans , Male , Hearing Loss, Noise-Induced , Noise, Occupational , Occupational Exposure , Surveys and QuestionnairesABSTRACT
<p><b>OBJECTIVE</b>To explore the variations of gene C in hepatitis B viruses between hepatitis B patients and healthy carriers, and provide experimental evidences for analysis of virus gene mutations acting on the virus material science and response of the body to the virus.</p><p><b>METHODS</b>The virus DNA load in hepatitis B patients and healthy blood donors was investigated by real-time polymerase chain reaction (PCR). Gene sequence analysis was taken to detect gene polymorphism, and all the success samples were compaired with standard strain by DNAstar.</p><p><b>RESULTS</b>(1)G Compared with standard strain, C region in all samples had mutations, there were 31 mutations in at least 2 samples (3 mutations in gene PreC and 28 mutations in gene C), including 9 missense mutations, 1 chain termination mutation and 21 synonymous mutation. Mutations nt 1827 c-->a and nt 2221 c-->t existed in all the samples, and most samples had 6 synonymous mutations. Four hepatitis B patients had mutation nt1896 g-->a, and another 4 patients had 2 mutations, namely, S87G and I97F (or 197L) in HBcAg CTL recognition episome. (2) The success ratio of amplification and sequencing of HBV DNA was closely associated with its copy numbers. In the present study, copy numbers of HBV DNA which were successfully amplified and sequenced were almost more than 40 193/ml.</p><p><b>CONCLUSIONS</b>HBV genome were easily affected by nucleotide mutations, 2 residues had mutations in gene of C region, which is firstly reported, suggesting these mutations may be geographical restricted. Mutations in gene of C region may either change the structure and function of HBeAg and HBcAg, which may further induce the escape of immune clearance for HBV or influence the detection of HBsAg or HBeAg, which may creat new problems for the prevention, diagnosis and treatment of hepatitis B.</p>
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Female , Humans , Male , Hepatitis B , Virology , Hepatitis B Core Antigens , Genetics , Hepatitis B virus , Genetics , Mutation , Polymorphism, GeneticABSTRACT
<p><b>OBJECTIVE</b>to evaluate the clinical outcomes of transpedicular lumbar wedge resection osteotomy in treating adult idiopathic scoliosis.</p><p><b>METHODS</b>twenty-five adult idiopathic scoliosis patients treated with transpedicular lumbar wedge resection osteotomy from July 2001 to November 2007 were included, among whom 18 were female and 7 were male. Nine of 25 were with double major curve in thoracic and thoracolumbar/lumbar spine, and 16 were with single curve in thoracolumbar/lumbar spine. The average age was 35 years (29 - 48 years) at operation. Osteotomy were performed at T(11), T(12), L(1) or L(2). The motion evoked potential monitoring system and awaking test were used during surgery. The preoperative, postoperative immediately and latest standing posteroanterior and lateral radiographs were reviewed.</p><p><b>RESULTS</b>all patients were operated successfully. The average operation time was 274 min (range, 220 - 380 min) and the average blood loss were 2328 ml (range, 1500 - 5000 ml). The average coronal Cobb angle of all patients in thoracolumbar/lumbar curves was 88° (range, 70° - 121°) before operation, which was corrected to 43° (range, 35° - 70°). The coronal correction rate was 44%. The average kyphosis angle of all in thoracolumbar/lumbar curves was 63° (range, 50° - 90°) before operation, which was corrected to 10° (range, -40° - 21°). The sagittal correction rate was 86%. Nerve root injury occurred in 3 of all patients who complained about postoperative radicular pain. No spine cord injury, delayed paralysis, infection and instrumentation failure were found. With a follow-up of 2 - 4 years, no correction loss or decompensation happened. The back pain existing before operation was relieved in large measure. The cosmetic appearance were all promoted significantly.</p><p><b>CONCLUSIONS</b>the transpedicular thoracolumbar/lumbar wedge osteotomy is efficient and safe in the correction of adult idiopathic scoliosis. The correction is much better on the sagittal plane than that on the coronal plane.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Follow-Up Studies , Osteotomy , Methods , Retrospective Studies , Scoliosis , General Surgery , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To determine the effects of berberine hydrochloride on the expressions of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in periodontal tissues in rat periodontitis model.</p><p><b>METHODS</b>Based on the successful rat periodontitis model, the experimental rats were randomized into different groups followed by oral treatment with berberine hydrochloride for 1, 2, 3, 4 weeks and then the rats were sacrificed and analyzed. Pathological assay and HE staining were used to detect the general conditions and pathological changes of rat periodontal tissues. And immunohistochemical staining was conducted to determine the expressions of IL-1beta and TNF-alpha in rats periodontitis model periodontal tissues.</p><p><b>RESULTS</b>The levels of IL-1beta and TNF-alpha in the periodontitis tissues were significantly higher than that in the control group. Treatment with berberine hydrochloride decreased the levels of IL-1beta and TNF-alpha in periodontitis tissues (P<0.05). Moreover, the general conditions and pathological changes in the control group and groups treated with berberine hydrochloride were much better than that in periodontitis groups.</p><p><b>CONCLUSION</b>Berberine hydrochloride inhibited the expression of IL-1beta and TNF-alpha in periodontal tissues in rats periodontitis model and promoted the regeneration of the periodontal tissues. This study suggested that berberine hydrochloride may have potential clinical application.</p>
Subject(s)
Animals , Male , Rats , Berberine , Interleukin-1beta , Periodontitis , Tumor Necrosis Factor-alphaABSTRACT
<p><b>OBJECTIVE</b>To study the low back pain(LBP) and its cause on female workers in flat-grained veneer wood industry.</p><p><b>METHODS</b>Bending posture was analyzed by observation and the prevalence of low back pain was investigated by physical examination and questionnaire among 299 female workers.</p><p><b>RESULTS</b>The prevalence of fatigue compliant in selecting, remending and sticking workers was 68.8%, 66.7% and 59.0%, respectively, which mainly involved in the part of low back. The prevalence of low back pain in selection (53.8%) and remending (58.7%) workers was higher than that in sticking workers (30.1%), which was in accordance with the tenderness between L4/L5 or L5/L6 and on the psoas major. Posture analysis indicated that the biggest bending range of selecting and remending workers (80 degrees ) was larger than that of sticking workers (60 degrees ), as well as the daily bending times[(4396+/-817), (1696+/-286), (1094+/-476)] and the time they kept bending[(6.5+/-0.6), (6.2+/-1.3), 4.5+/-0.9) h].</p><p><b>CONCLUSION</b>Bending posture is common among female workers especially those who work in selecting and remending and might be the major causes for the high prevalence of LBP in flat-grained veneer wood industry.</p>
Subject(s)
Adult , Female , Humans , Low Back Pain , Epidemiology , Occupational Diseases , Epidemiology , Posture , Prevalence , Risk Factors , Surveys and Questionnaires , WoodABSTRACT
<p><b>OBJECTIVE</b>To investigate HMGB-1 expression and its extracellular release of cultured primary hepatic parenchymal cells (HC) and Kupffer cells (KC) that were induced by lipopolysaccharides (LPS).</p><p><b>METHODS</b>Primary hepatic parenchymal cells and Kupffer cells were cultured in flasks, and some cells were treated with 500 microg/L LPS for 24 hours (induced group) and some were not treated with LPS and served as controls. All of the cells were repeatedly frozen-thawed, and the expression levels of HMGB1-mRNA and HMGB1 proteins were detected by semi-quantitative RT-PCR and Western blot respectively. Then HC and KC were subcultured in 24-well culture plates for 6 h, 12 h, 24 h and 48 h, and the HMGB1 protein in culture fluids was detected by Western blot at each time point.</p><p><b>RESULTS</b>Compared with the cells in the control group, the expression levels of HMGB1-mRNA in the induced group were significantly increased in both HC and KC at 24 h (t=31.32 and 45.90, P<0.05) and the protein levels of HMGB1 showed the same results (t=46.19 and 38.44, P<0.05). There was a small quantity of HMGB1 protein in the culture fluids of two control groups and the induced group of HC. However the HMGB1 protein in the induced group of KC were obviously increased with prolonged culture time (F=42.74, P<0.05). Compared with the control group, the level of HMGB1 protein in the induced group of KC was not increased at 6 h (t=9.57, P>0.05) but was significantly increased at 12 h, 24 h and 48 h (t=21.95, 32.39, 44.16, respectively P<0.05).</p><p><b>CONCLUSION</b>LPS could increase HMGB1 expression of HC and KC and HMGB1 release from KC, but not from HC. The results suggest that KC play an important role in triggering inflammation and liver injury.</p>