OBJECTIVETo establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.

METHODSThe femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde, decalcified, dehydrated, embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type II collagen, MMP13, Sox9 and ADAMTS5 were analyzed by immunofluorescence.

RESULTSBoth the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group, which indicated that cartilage became degenerated. The expression levels of both type H collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.

CONCLUSIONThe model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro.

Animals ; Cartilage Diseases ; genetics ; metabolism ; Collagen Type II ; genetics ; metabolism ; Disease Models, Animal ; Femur Head ; metabolism ; Humans ; In Vitro Techniques ; Interleukin-1beta ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; genetics ; metabolism

AIM: To provide the morphological evidence for the compound carbomer building high intraocular pressure model successfully.? METHODS: Totally 50 SD rats were divided into experimental group 40 rats, blank group 10 rats by random number method. The rats in experimental group were randomly divided into 3 groups after the model was successful. Experimental model of high intraocular pressure was sacrificed at 1, 2 and 3wk to observe its pathological structure change and ultrastructure change.? RESULTS: The high intraocular pressure of experimental model in 1, 2 and 3wk all showed the optic nerve and retinal damage. It was to see the optic nerve axon disappear, disorder arrangement of myelin sheath, periodic dissolve or demyelinating degeneration, glial cell proliferation. It showed the cells disordered arrangement of retina, the outer nuclear layer became thick, the inner and outer plexiform layer become thick, the kernel layer became thick, air bubbles, the numbers of ganglion cells reduced, ganglion cells and nerve fiber layer edema, microglia proliferation, vascular membrane capillaries expansion, inflammatory cells appearing. It was to see the retinal ganglion cells layer with microglia proliferation under the electron microscope, ganglion cells structure fuzzy, organelles structures disappear, cell apoptosis, mitochondrial swelling, cytoplasm vacuoles degeneration, membrane plate of outer segment fracture or dissolved. And the damage degree was proportional to the forming time of high intraocular pressure.? CONCLUSION: The morphology change of high intraocular pressure model about the retin and optic nerve proves that it is successful building the model through anterior chamber injection of compound carbomer solution.

OBJECTIVETo analyze the impact of depletion of the twin arginine translocation (TAT) system on virulence and physiology of Yersinia enterocolitica for a better understanding of its pathogenicity.

METHODSWe constructed a DeltatatC::SpR mutant of Yersinia enterocolitica by P1 phage mediated transduction using Escherichia coli K-12 DeltatatC::SpR strain as a donor.

RESULTSA P1-mediated genetic material transfer was found between the two species of enterobacteria, indicating a great potential of acquisition of antibiotic resistance in emergency of a new threatening pathogen by genetic material exchanges. Periplasmic trimethylamine N-oxidase reductase activity was detected in the wild type Y. enterocolitica strain and translocation of this enzyme was completely abolished by the DeltatatC::SpR mutation. In addition, the DeltatatC::SpR mutation showed a pleiotropic effect on the metabolism of Y. enterocolitica. However, the tat mutation did not seem to affect the mobility and virulence of Y. enterocolitica under the conditions used.

CONCLUSIONUnlike other pathogenic bacteria studied, the TAT system of Y. enterocolitica might play an important role in the pathogenic process, which is distinct from other pathogens, such as Pseudomonas aeruginosa and enterohemorrhagic E. coli O157:H7.

Drug Resistance, Microbial ; genetics ; Genes, Bacterial ; Mutation ; Oxidoreductases Acting on CH-NH Group Donors ; metabolism ; Transduction, Genetic ; Virulence ; Yersinia enterocolitica ; enzymology ; genetics ; metabolism ; pathogenicity

This study was aimed to explore the clinical significance of monitoring level of minimal residual disease (MRD) at different time point in B-lineage childhood acute lymphoblastic leukemia (B-ALL). Two hundred and six children with B-ALL were enrolled in this study from Augest 2008 to September 2011 in our hospital. MRD levels were detected by flow cytometry at day 15, 33 and week 12 after initial chemotherapy. The event-free survival (EFS) for patients based on MRD levels measured at different stages of chemotherapy were compared by Kaplan Meier analyses. The results showed that out of 206 cases 196 cases achieved complete remission (CR) after induction therapy (CR rate 95.1%), the 1- and 3-year EFS rate were (92.7 ± 1.8)% and (78.7 ± 3.7)%, respectively, and the 3-year EFS rate was (85.6 ± 4.9)% in standard risk group, (82.1 ± 5.8)% in intermediate risk group and (58.1 ± 9.2)% in high risk group, there was significant statistical difference between above mentioned 3 groups (P < 0.001). The MRD analysis at different time points showed that the higher the MRD level, the lower the 3-year EFS rate of children with ALL, in which the 3-year EFS rate of MRD ≥ 10(-2) at day 15, MRD ≥ 10(-3) at day 33 and MRD ≥ 10(-3) at week 12 were significantly lower. The MRD ≥ 10(-3) at week 12 was proven to be an independent predictor by multivariate Cox proportional-hazards regression model. The 3-year EFS rate for patients with MRD < 10(-3) and MRD ≥ 10(-3) at week 12 were (86.3 ± 4.1)% vs (55.8 ± 9.1)% (P < 0.05); 8 relapsed among 98 cases with negative MRD (MRD < 10(-4)) at day 33, 19 relapsed among 108 cases with positive MRD at day 33 between the two groups for recurrence rate has significant difference (P < 0.05). It is concluded that dynamically monitoring MRD by multi-parameter flow cytometry can precisely evaluate treatment response, judge treatment outcome and predict relapse in childhood B-ALL. The MRD 10(-2) at day 15, MRD 10(-3) at day 33 and MRD 10(-3) at week 12 should be considered as the best cut-off. MRD ≥ 10(-3) at week 12 was proven to be an independent factor of poor prognosis.
Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Infant ; Male ; Neoplasm, Residual ; diagnosis ; therapy ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; therapy ; Prognosis ; Retrospective Studies

OBJECTIVETo study the status and problems of complementary feeding behavior for infants aging between 6 - 24 month-old in urban and rural areas of Chengdu in 2011.

METHODSA total of 1283 infants aging between 6 - 24 month-old, including 601 from urban area and 682 from rural area, were randomly selected by stratified cluster random sampling method, and divided into three groups as 6 - 8, 9 - 11 and 12 - 24 month-old groups, between March and April in 2011. Information of complementary feeding behavior were collected by questionnaire, and then assessed, quantified and compared by standards. The evaluation system was established (the total score was 24), and was used to assess among children aging 6 to 24 month-old in urban and rural areas in Chengdu.

RESULTSThe average score for infants in Chengdu was 18.6 ± 2.5, accounting for 77.6% of the total score. The scores of food selection and preparation were highest (90.7% of total score) among the behavior, and the scores of caregiver's behavior, infants behavior, feeding environment and environmental hygiene were low (about 75.2% - 77.6% of total score). The average score in urban and rural groups were separately 19.8 ± 2.1 and 17.6 ± 2.4, the score in urban group was higher than that in rural group (P < 0.05), occupying total score 82.3%, 73.4%, respectively. The average score in 6 - 8 month, 9 - 11 month and 12 - 24 month groups were separately 18.9 ± 2.3, 19.1 ± 2.4, and 18.2 ± 2.6, occupying total score 78.9%, 79.4% and 75.7%, respectively. The score in 6 - 8 and 9 - 11 month groups were higher than that in 12 - 24 month group (P < 0.01). The pass rate (above 60% of total score) was 94.2% and excellent rate (above 80% of total score) was 44.8% in Chengdu. The excellent rate in urban and rural groups were separately 65.7% and 26.4% (χ(2) = 199.825, P < 0.01).

CONCLUSIONThe complementary feeding behavior for infants was not optimistic in Chengdu, and the behavior was even worse in rural areas and among elder infants groups.

Child, Preschool ; China ; Feeding Behavior ; Food Preferences ; Humans ; Infant ; Infant Food ; Infant Nutritional Physiological Phenomena ; Rural Population ; Surveys and Questionnaires ; Urban Population

Objective To investigate the effect ofbiglycan (BGN) on neural apoptosis in mice with early brain injury (EBI) after subarachnoid hemorrhage (SAH).Methods SAH models were induced by endovascular perforation in young male C57BL/6J mice.(1) Totally,48 mice were randomly divided into sham-operated group,SAH 6 h group,SAH 12 h group,SAH 24 h group,SAH 48 h group,and SAH 72 h group (n=8);the BGN protein and mRNA expressions were detected by Western blotting and real-time quantitative PCR (qRT-PCR).(2) Totally,16 mice were randomly divided into sham-operated group and SAH 48 h group (n=8);immunofluorescent double staining was conducted to explore the BGN expression in the neurons of brain tissues.(3) Totally,24 mice were randomly divided into sham-operated group,sham+control lentivirus group,and sham+BGN lentivirus group (n=8);BGN lentiviral vector and control lentivirus were administered intracerebroventricularly 7 d before sham-operation;qRT-PCR was performed to explore the BGN mRNA expression.(4) Totally,48 mice were randomly divided into sham-operated group,SAH+control lentivirus group,and SAH+BGN lentivirus group (n=16);BGN lentiviral vector and control lentivirus were administered intracerebroventricularly 7 d before SAH;neurological scores were detected by modified Garcia scale and beam balance tests;TUNEL was used to detect the neuronal apoptosis,and Western blotting was performed to explore the expressions of nuclear transcription factor kappa B (NF-κB) and phosphorylated-(p-) NF-κB.Results (1) Mice in the SAH 48 h group had the highest BGN protein and mRNA expressions,which showed statistical differences as compared with the sham-operated group (P＜0.05).(2) A majority of BGN expressions were detected in the neurons 48 h after SAH.(3) The sham+BGN lentivirus group had statistically lower BGN mRNA expression than the sham+control lentivirus group (P＜0.05).(4) As compared with those in the SAH+control lentivirus group,both scores of modified Garcia scale and beam balance tests were significantly higher in SAH+BGN lentivirus group (6.125±1.246 vs.13.000±1.309;1.125±1.126 vs.2.875±0.835),and neural apoptosis ratio and ratio of p-NF-κB/NF-κB were significantly lower in the SAH+BGN lentivirus group (51.950％±11.166％ vs.31.938％±7.705％;1.161±0.156 vs.0.886±0.142,P＜0.05).Conclusion Inhibition of BGN can effectively reduce neuronal apoptosis in mice with EBI after SAH,and attenuate neurological deficits.

OBJECTIVETo determine the incidence and risk factors of HCV infection among heroin addicts who were receiving methadone maintenance treatment(MMT)in Dehong prefecture, Yunnan province.

METHODSAll heroin addicts who were HCV negative at the initiation of MMT in June 2005 through March 2012, in Dehong prefecture, were included in this cohort analysis. HCV incidence was calculated and related risk factors determined by using Cox proportional hazard regression model.

RESULTSA total of 2390 MMT clinic attendants were qualified for this cohort study by March 2012. 731(30.6%) of them had never received any follow-up HCV testing so were recognized as loss to follow-up. The other 1659 (69.4%) participants had received at least one follow-up HCV testing and were observed for a total of 3509.12 person-years(py). During this period 99 new HCV infections or HCV sero-converters were identified. The overall HCV incidence was 2.82/100 py and was 3.62/100 py for 2006, 5.36/100 py for 2007, 6.71/100 py for 2008, 2.56/100 py for 2009, 1.90/100 py for 2010, and 0.44/100 py for 2011, respectively. Results from multiple regression analysis, using Cox proportional hazard model, indicated that after controlling for confounding variables, those who were unemployed, being injecting drug users(IDUs)or HIV positive at entry into the MMT program were more likely to be newly infected with HCV or HCV sero-converted during the follow-up period than those who were peasants, non-IDUs or HIV negative at entry into the MMT program(HR = 2.02, 95% CI:1.18-3.48; HR = 9.05, 95% CI:5.49-14.93; HR = 2.12, 95% CI: 1.37-3.56), respectively.

CONCLUSIONThe incidence of HCV infection among MMT clinic attendants was decreasing since 2009 in Dehong prefecture. Those who were unemployed, injecting drug users and HIV positive were at higher risk of HCV infection.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Cohort Studies ; Drug Users ; Female ; Hepatitis C ; epidemiology ; Humans ; Incidence ; Male ; Methadone ; therapeutic use ; Risk Factors ; Substance-Related Disorders ; drug therapy ; epidemiology ; virology

Objective To determine the incidence and risk factors of HIV infection among heroin addicts receiving methadone maintenance treatment(MMT)in Dehong prefecture,Yunnan province.Methods All heroin addicts who were HIV negative at the initiation of MMT in June 2005 and through June 2011,in Dehong prefecture were included in the cohort analysis.HIV incidence was calculated and related risk factors determined by using Cox proportional hazard regression model.Results A total of 3154 MMT clinic attendants were qualified for this cohort study.By June 2011,1023(32.4％)of them had never received any follow-up HIV testing so were thus referred as loss to follow-up.The other 2131(67.6％)members had received at least one follow-up HIV testing and were observed for a total of 4615.86 person-years.During the period,22 new HIV infections or seroconverters were identified,making the overall HIV incidence as 0.48/100 person-years.The HIV incidence was higher among those who were unemployed,never married,self-reported being injecting drug users(IDUs)and HCV positive at entry into the MMT program.None of those who were always negative on follow-up-urine-testing of morphine was discovered as HIV newly infected during the follow-up period.Data from multiple regression analysis under Cox proportional hazard model indicated that after controlling for confounding variables,non-IDUs at the entry point for the MMT program,were less likely to be HIV newly-infected or seroconverted than IDUs(HR=0.29,95％CI:0.11-0.76).Conclusion MMT prograqm in Dehong prefecture was demonstrated to be fairly effective in reducing HIV transmission through drug use.Those HIV negative attendants at the MMT clinic who were IDUs or keep using drugs during the treatment,were at higher risk of HIV seroconvertion.More efforts were needed to improve the follow-up and HIV testing programs for the MMT clinic attendants.

Objective:To develop a tetramer probe targeting fibroblast activation protein (FAP), named 1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid (DOTA)-4P(FAP inhibitor (FAPI)) 4, evaluate its biodistribution and PET image in FAP-positive-tumor bearing nude mice, and explore its feasibility as a novel radio-regent for treatment of FAP-positive tumor. Methods:FAP tetramer probe was constructed on the FAPI-46 motif with four mini-polyethylene glycol (PEG)(PEG 3) spacers between the four FAPI motifs, denoted as 4P(FAPI) 4. DOTA was used as the chelator for radiolabeling with 68Ga and 177Lu. The FAP binding characteristics were test by in vitro cell competitive binding experiment. Small-animal PET, in vivo biodistribution, and radionuclide targeting therapy were performed in HT-1080-FAP tumor bearing nude mice ( n=39). Independent-sample t test was performed to analyze tumor uptake data, and two-factor repeated measures analysis of variance was utilized to compare tumor volume data in radioactive isotope therapy. Results:Cell experiment showed that FAPI-tetramer and FAPI-monomer had similar half maximal inhibitory concentration values (3.29 and 2.15 nmol/L). 68Ga/ 177Lu radiolabeled FAPI-tetramer had better tumor uptake and retention than FAPI-monomer in small-animal PET and in vivo biodistribution experiment, with the tumor uptake for 177Lu-DOTA-4P(FAPI) 4 and 177Lu-FAPI-46 at 48 h of (18.72±1.32) vs (2.72±1.20) percentage activity of injection dose per gram of tissue (%ID/g) ( t=15.55, P<0.001). 177Lu-DOTA-4P(FAPI) 4 group showed best anti-tumor efficacy compared with 177Lu-FAPI-46 and control group in radionuclide targeting therapy. On the 2nd day after the start of treatment, the tumor volume in the tetramer treatment group was significantly smaller than that in the control group (mean difference 67.19 mm 3, P=0.049); on the 14th day after the start of treatment, the tumor volume in the tetramer treatment group was significantly smaller than that in the monomer treatment group (mean difference 414.33 mm 3, P=0.005). Conclusion:FAPI-tetramer can improve tumor uptake and retention ability compared with FAPI-46, and 177Lu-DOTA-4P(FAPI) 4 can be a promising radio-agent for FAP-positive tumor therapy.

OBJECTIVE: To systematically evaluate the clinical efficacy of high-quality direct anterior approach (DAA) and other approaches for the treatment of elderly patients with femoral neck fracture. METHODS: Literatures published in English or Chinese about the direct anterior approach and other approaches for hemiarthroplasty in femoral neck fracture were searched on Cochrane Library, PubMed, EMBASE, Web of science, Wanfang, CNKI databases from their establishment to May 2019. According to the inclusion and exclusion criteria, two researchers independently screened the literatures, and extracted the data. The quality of RCT were evaluated by Cochrane Risk of Bias Assessment Tool, and non-RCT were evaluated by the NOS scale. Meta-analysis was performed using the RevMan 5.3 software. RESULTS: A total of 9 articles were included with 901 cases, in which 429 cases used DAA, and 472 used other approaches. DAA had a significantly lower dislocation rate compared to subgroup of posterior and posterolateral approach [=0.19, 95%CI (0.06, 0.61), =0.005]. No significant differences were found between DAA group and subgroup of direct lateral and anterolateral approach[=1.08, 95%CI(0.20, 5.76), =0.93]. Also there were no relevant differences between the DAA group and control in infection rate[=1.07, 95%CI(0.47, 2.43), =0.88], perioperative fracture rate[=0.95, 95%CI(0.36, 2.50), =0.92], re operation rate[=0.76, 95%CI(0.30, 1.89), =0.55], overall complication rate [=0.88, 95%CI (0.63, 1.22), =0.44], mortality [=1.33, 95%CI (0.84, 2.11), =0.23], operative time[MD=1.43, 95%CI(-5.85, 8.71), =0.70]. CONCLUSION The current evidenceindicates that the DAA was associated with a significantly lower dislocation rate compared to posterior capsular approaches for hemiarthroplasty. There was no significant difference in dislocation rate with the lateral and anterolateral approach.
Aged ; Antiviral Agents ; Arthroplasty, Replacement, Hip ; Femoral Neck Fractures ; surgery ; Hemiarthroplasty ; Hepatitis C, Chronic ; Humans ; Reoperation ; Treatment Outcome