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Objective To investigate the level of deltamethrin resistance and mutation sites in the sodium iron channel gene in Rhipicephalus microplus in Huaihua City, Hunan Province, and to examine the correlation between deltamethrin resistance and mutation sites in the sodium iron channel gene in Rh. microplus. Methods Rh. microplus was sampled from multiple yellow cattle farms in Huaihua City, Hunan Province from June to September 2022, and the level of resistance to deltamethrin was determined in ticks using the adult immersion test. The sodium iron channel domain III gene was amplified in deltamethrin-resistant and wild-type Rh. microplus using PCR assay. Following sequencing and sequence alignment, mutation sites were detected in bases. The sodium iron channel domain III gene in Rh. microplus was translated, and the signal peptide, transmembrane domain, and phosphorylation and glycosylation sites were detected in amino acid sequences. The tertiary structures of the sodium iron channel domain III protein of deltamethrin-resistant and wild-type Rh. microplus were deduced and compared, and the association be tween mutation sites in bases and resistance to deltamethrin was examined in Rh. microplus according the level of deltamethrin resistance, sequence alignment and protein tertiary structure. Results The median (LC50) and 95% lethal concentrations (LC95) of deltamethrin were 121.39 mg/L and 952.61 mg/L against Rh. microplus, with a resistance factor of 9.24 and level II resistance. The sequence of the sodium ion channel domain III gene was 1 010 bp in size, and mutation sites were detected in two neighboring bases in the sequence of the sodium ion channel domain III gene in deltamethrin-resistant Rh. microplus. Although no signal peptides were found in the sodium iron channel domain III protein of deltamethrin-resistant or wild-type Rh. microplus, 6 trans-membrane domains, 42 phosphorylation sites and 8 glycosylation sites were identified, with a significant difference in the tertiary structure of the sodium iron channel domain III protein between deltamethrin-resistant and wild-type Rh. microplus. Conclusions Level II resistance to deltamethrin is detected in Rh. microplus in Huaihua City, Hunan Province, and two mutation sites that correlate with the emergence of deltamethrin resistance are identified in the sequence of the sodium iron channel domain III gene in deltamethrin-resistant Rh. microplus.
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ObjectiveTo describe the epidemiological characteristics of confirmed COVID-19 cases in Shangrao City， and provide scientific evidence for the development of prevention and control strategies. MethodsEpidemiological characteristics of 123 confirmed cases， such as time， sex， age， occupations， area， and source of infection， were analyzed by combining epidemiological investigation with laboratory testing. ResultsOf the 123 confirmed cases， 34 were imported cases and 89 were local cases. The sex ratio was 1.32∶1. Average time interval of the COVID-19 was （18.4±4.4） days， with a median time interval between the onset and final diagnosis of 5 days. There were 19 clustering cases， of which 89.5% were family clusters and 77.3% occurred in families. Majority of the cases were common type. The main source of infection was travelers from the epidemic areas. ConclusionHealth monitoring of travelers from epidemic areas and strict quarantine of close contacts are the main measures to control the spread of the COVID-19 epidemic.
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ObjectiveTo describe the epidemiological characteristics of confirmed COVID-19 cases in Shangrao City， and provide scientific evidence for the development of prevention and control strategies. MethodsEpidemiological characteristics of 123 confirmed cases， such as time， sex， age， occupations， area， and source of infection， were analyzed by combining epidemiological investigation with laboratory testing. ResultsOf the 123 confirmed cases， 34 were imported cases and 89 were local cases. The sex ratio was 1.32∶1. Average time interval of the COVID-19 was （18.4±4.4） days， with a median time interval between the onset and final diagnosis of 5 days. There were 19 clustering cases， of which 89.5% were family clusters and 77.3% occurred in families. Majority of the cases were common type. The main source of infection was travelers from the epidemic areas. ConclusionHealth monitoring of travelers from epidemic areas and strict quarantine of close contacts are the main measures to control the spread of the COVID-19 epidemic.
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Objective To investigate the genetic variation of Eurytrema pancreaticum isolated from goats in Huaihua City, Hunan Province. Methods The partial sequence of mitochondrial cytochrome I (pcox1) and ribosomal 18S rRNA genes were amplified using a PCR assay in E. pancreaticum isolates from goats in Huaihua City, Hunan Province, and the PCR amplification products were sequenced. Then, the gene sequences were subjected to genetic variation and phylogenetic analyses. Results The sequences of the pcox1 and 18S rRNA genes were 430 bp and 1 857 bp in length in 18 E. pancreaticum isolates from goats in Huaihua City, Hunan Province, and there were 14 and 35 variation sites in pcox1 and 18S rRNA gene sequences, with intra-species genetic variations of 0 to 1.4% and 0 to 0.8%, respectively. The sequences of pcox1 and 18S rRNA genes had 99.0% to 99.8% and 99.5% to 99.8% homologies with those from E. pancreaticum Chinese strain recorded in the GenBank database. Consistent phylogenetic analysis results were found based on pcox1 and 18S rRNA genes. The 18 E. pancreaticum isolates from goats in Huaihua City were clustered into a clade with the known E. pancreaticum isolates registered in GenBank, and the clade with these 18 E. pancreaticum isolates was close to the clades with Eurytrema species and far from the clades with other trematodes. Conclusions The E. pancreaticum isolates from goats have a low genetic variation in Huaihua City, Hunan Province. Mitochondrial pcox1 and ribosomal 18S rRNA genes may serve as molecular markers for the studies on the genetic variation in goat-derived E. pancreaticum.
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Mitochondria, as the key passway of neuronal apoptosis after ischemia, is closely related to cerebral ischemia-reperfusion injury. Remote ischemic post-conditioning can alleviate cerebral ischemia-reperfusion injury, and its mechanism is related to alleviating mitochondrial injury and improving its dysfunction. In this paper, cytochrome C/caspase, mitophagy, mitochondrial ATP-sensitive K+ channel and mitochondrial permeability transitionpore were reviewed.
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Pericytes, endothelial cells (EC), astrocytes and extracellular space together constitute the blood-brain barrier. Pericytes and EC participate in the various regulations of blood-brain barriers through many mechanisms to maintain the stability of neurovascular units (NVU). The injury and repair of NVU involve a lot of signal transduction at molecular levels, and angiogenesis is primarily about the generation and maturation of EC and the supporting adhesion process of pericytes. This article briefly reviewed EC-related angiogenesis signaling pathways in pericytes after NVU ischemic injury.
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Pericytes, endothelial cells (EC), astrocytes and extracellular space together constitute the blood-brain barrier. Pericytes and EC participate in the various regulations of blood-brain barriers through many mechanisms to maintain the stability of neurovascular units (NVU). The injury and repair of NVU involve a lot of signal transduction at molecular levels, and angiogenesis is primarily about the generation and maturation of EC and the supporting adhesion process of pericytes. This article briefly reviewed EC-related angiogenesis signaling pathways in pericytes after NVU ischemic injury.
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OBJECTIVE@#To investigate the diagnostic value of Hepcidin as a sepsis biomarker in critically ill adults.@*METHODS@#An observational study was conducted. The patients with suspected or proven infection admitted to intensive care unit (ICU) of Zhoupu Hospital Affiliated to Shanghai University of Medicine and Health Sciences from March 2016 to November 2017 were enrolled. According to the third international consensus definitions for sepsis and septic shock (Sepsis-3), the patients were divided into non-sepsis group and sepsis group, and the septic patients were subdivided into general sepsis subgroup and septic shock subgroup according to the severity of disease. The differences in serum Hepcidin, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), procalcitonin (PCT), C-reactive protein (CRP), white blood cell (WBC), neutrophil granulocytes (NEUT) and lactic acid (Lac) within 1 hour after ICU admission between non-sepsis and sepsis groups and among the sepsis subgroups were compared. The acute physiology and chronic health evaluation II (APACHE II) within 24 hours after ICU admission and sequential organ failure score (SOFA) were recorded, and the mortality rate was followed up for 28 days. Receiver operation characteristic curve (ROC) was used to evaluate and compare the diagnostic value of Hepcidin and PCT, CRP, WBC for sepsis. Logistic regression model was used to estimate the association between Hepcidin and sepsis. Spearman correlation analysis was used to analyze the correlation between Hepcidin and other parameters of sepsis patients.@*RESULTS@#A total of 183 patients were enrolled, 93 in the non-sepsis group and 90 in the sepsis group (48 with general sepsis and 42 with septic shock). (1) The levels of Hepcidin, IL-6, TNF-α, PCT, Lac in serum, and APACHE II and SOFA scores in the sepsis group were significantly higher than those in the non-sepsis group. ROC analysis showed that the area under the ROC curve (AUC) of Hepcidin and PCT for sepsis diagnosis were 0.865 [95% confidence interval (95%CI) = 0.807-0.911] and 0.848 (95%CI = 0.788-0.897), respectively, without statistical significance (Z = 0.443, P = 0.657). Furthermore, the AUC of Hepcidin for sepsis diagnosis was significantly higher than that of the conventional biomarkers CRP and WBC [AUC was 0.530 (95%CI = 0.455-0.604) and 0.527 (95%CI = 0.452-0.601), respectively] with statistical significance (both P < 0.01). When Hepcidin > 54.00 μg/L, its sensitivity for sepsis diagnosis was 95.56%, specificity was 66.67%, positive and negative predictive value was 73.51% and 93.94%, respectively. Parallel test was conducted for combination of Hepcidin and PCT, which showed that the AUC was 0.885, and the sensitivity and negative predictive value was significantly improved to 98.96% and 98.36%, respectively. Logistic regression analysis demonstrated that after adjusted for PCT, Hepcidin > 54.00 μg/L was also associated with sepsis independently, with odds ratio (OR) of 1.011 (95%CI = 1.008-1.015, P < 0.001), indicating that Hepcidin and PCT were not completely overlapped in the diagnosis of sepsis. (2) With the increase in infection severity, serum Hepcidin, PCT, IL-6, TNF-α, Lac, APACHE II, SOFA score and 28-day mortality all showed an increasing trend in patients. There was a significantly positive correlation between Hepcidin and IL-6, TNF-α, PCT, APACHE II, and SOFA in the sepsis patients (r value was 0.526, 0.449, 0.591, 0.359, and 0.374, respectively, all P < 0.01), but no correlation was found between Hepcidin and Lac (r = 1.104, P > 0.05).@*CONCLUSIONS@#Serum Hepcidin is a useful biomarker for the diagnosis of sepsis, and it is correlated to the severity of the sepsis. The combination of Hepcidin and PCT can improve the accuracy of diagnosis of sepsis.@*CLINICAL TRIAL REGISTRATION@#China Clinical Trial Registration Center, ChiCTR-DDD-16008522.
Subject(s)
Adult , Humans , Biomarkers , C-Reactive Protein , Calcitonin , Calcitonin Gene-Related Peptide , China , Critical Illness , Hepcidins , Prognosis , Protein Precursors , ROC Curve , SepsisABSTRACT
Objective@#To investigate the diagnostic value of Hepcidin as a sepsis biomarker in critically ill adults.@*Methods@#An observational study was conducted. The patients with suspected or proven infection admitted to intensive care unit (ICU) of Zhoupu Hospital Affiliated to Shanghai University of Medicine & Health Sciences from March 2016 to November 2017 were enrolled. According to the third international consensus definitions for sepsis and septic shock (Sepsis-3), the patients were divided into non-sepsis group and sepsis group, and the septic patients were subdivided into general sepsis subgroup and septic shock subgroup according to the severity of disease. The differences in serum Hepcidin, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), procalcitonin (PCT), C-reactive protein (CRP), white blood cell (WBC), neutrophil granulocytes (NEUT) and lactic acid (Lac) within 1 hour after ICU admission between non-sepsis and sepsis groups and among the sepsis subgroups were compared. The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) within 24 hours after ICU admission and sequential organ failure score (SOFA) were recorded, and the mortality rate was followed up for 28 days. Receiver operation characteristic curve (ROC) was used to evaluate and compare the diagnostic value of Hepcidin and PCT, CRP, WBC for sepsis. Logistic regression model was used to estimate the association between Hepcidin and sepsis. Spearman correlation analysis was used to analyze the correlation between Hepcidin and other parameters of sepsis patients.@*Results@#A total of 183 patients were enrolled, 93 in the non-sepsis group and 90 in the sepsis group (48 with general sepsis and 42 with septic shock).① The levels of Hepcidin, IL-6, TNF-α, PCT, Lac in serum, and APACHEⅡand SOFA scores in the sepsis group were significantly higher than those in the non-sepsis group. ROC analysis showed that the area under the ROC curve (AUC) of Hepcidin and PCT for sepsis diagnosis were 0.865 [95% confidence interval (95%CI) = 0.807-0.911] and 0.848 (95%CI = 0.788-0.897), respectively, without statistical significance (Z = 0.443, P = 0.657). Furthermore, the AUC of Hepcidin for sepsis diagnosis was significantly higher than that of the conventional biomarkers CRP and WBC [AUC was 0.530 (95%CI = 0.455-0.604) and 0.527 (95%CI = 0.452-0.601), respectively] with statistical significance (both P < 0.01). When Hepcidin > 54.00 μg/L, its sensitivity for sepsis diagnosis was 95.56%, specificity was 66.67%, positive and negative predictive value was 73.51% and 93.94%, respectively. Parallel test was conducted for combination of Hepcidin and PCT, which showed that the AUC was 0.885, and the sensitivity and negative predictive value was significantly improved to 98.96% and 98.36%, respectively. Logistic regression analysis demonstrated that after adjusted for PCT, Hepcidin > 54.00 μg/L was also associated with sepsis independently, with odds ratio (OR) of 1.011 (95%CI = 1.008-1.015, P < 0.001), indicating that Hepcidin and PCT were not completely overlapped in the diagnosis of sepsis. ② With the increase in infection severity, serum Hepcidin, PCT, IL-6, TNF-α, Lac, APACHEⅡ, SOFA score and 28-day mortality all showed an increasing trend in patients. There was a significantly positive correlation between Hepcidin and IL-6, TNF-α, PCT, APACHEⅡ, and SOFA in the sepsis patients (r value was 0.526, 0.449, 0.591, 0.359, and 0.374, respectively, all P < 0.01), but no correlation was found between Hepcidin and Lac (r = 1.104, P > 0.05).@*Conclusions@#Serum Hepcidin is a useful biomarker for the diagnosis of sepsis, and it is correlated to the severity of the sepsis. The combination of Hepcidin and PCT can improve the accuracy of diagnosis of sepsis.@*Clinical trial registration@#China Clinical Trial Registration Center, ChiCTR-DDD-16008522.
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[Objective] To investigatethe correlation between blood flow signal on three dimensional time-of-flight magnetic resonance angiography (3D-TOF-MRA) and short-term outcome in patients with symptomatic middle cerebral artery (MCA) severe stenosis or MCA occlusion.[Methods] We retrospectively reviewed consecutive patients with symptomatic MCA severe stenosis or MCA occlusion.General information,clinical data and cranial imaging data were collected.Characteristics of blood flow signal on 3D-TOF-MRA for each patient were analyzed,which included:(1) blood flow signal of MCA distal to stenosis/occlusion lesion;(2) laterality of posterior cerebral artery (PCA).The correlation between characteristics of blood flow signal and short-term outcome was analyzed.[Results] Three hundred and twenty-eight patients were included in this study.There were 154 patients with symptomatic MCA severe stenosis and the rest of them had symptomatic MCA occlusion.Poor blood flow signal of distal MCA independently correlated with poor shortterm outcome in patients with severe MCA stenosis.[Odds Ratio (OR) 0.32,95％ Confident Interval (CI) 0.14～0.72].PCA laterality was not related with short-term outcome in these patients (OR,2.28,95％ CI,0.85～6.15).PCA laterality independently correlated with poor short-term outcome in patients with MCA occlusion.(OR,3.54,95％ CI,1.32～ 9.78).Blood flow signal of distal MCA was not related with short-term outcome in these patients (OR,0.58,95％ CI,0.22～1.48).[Conclusion] Blood flow signal on 3D-TOF-MRA correlates with short-term outcome in patients with symptomatic MCA severe stenosis or occlusion but the characteristics differs between severe MCA stenosis and occlusion patients.Anterograde blood flow (blood flow signal of MCA distal to stenosis lesion) for patients with severe MCA stenosis and retrograde blood flow (PCA laterality) for patients with MCA occlusion correlates with shot-term outcome.
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Combined headspace solid-phase microextraction with gas chromatography-mass spectrometry (HS-SPME-GC-MS), this paper studied the volatiles of Artemisiae Argyi Folium that respectively collected from Kongdong, Huanglong, Ankang, Qichun and Tongbai，and then explored the influence of climatic factors on the main components of the volatiles. The results showed that 54, 52, 57, 47 and 55 compounds were respectively identified from the leaves from five regions, mainly consist of monoterpenes, sesquiterpenoids and their oxygen-containing derivatives, a few aldehydes, alkanes and benzenes. It is different that the volatiles of Artemisiae Argyi Folium that collected in different habitats, neither composition nor content. There are total 18 kinds of compounds that exist in all five leaves. Eucalyptol, camphor, terpinen-4-ol, eugenol and caryophyllene could be considered to be the main conponents because of their efficacy or toxicity and relatively high content. Grey correlation analysis shows that the annual lowest temperature and annual sunshine duration have a great influence on the content of eucalyptol and camphor; the effect of annual precipitation and annual accumulated temperature on the content of terpinen-4-ol and caryophyllene was relatively significant; while the annual sunshine duration and the annual highest temperature are the main influencing factors for eugenol. The research suggests that long-day and extreme temperature may be more conductive to the accumulation of volatiles. According to the laws of effect of climatic factors on volatiles, we can optimize the cultivation conditions and select suitable locality to improve the quality of Artemisiae Argyi Folium.
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Objective To study the intrinsic relationship between hemiplegic gait features and abnormal muscle strength of lower limbs, so as to elucidate the muscle strength causes of hemiplegic gait and provide recommendations for its clinical treatment. Methods Models of hemiplegic gait and normal gait were established for simulation, and the validity of the models was verified by comparing the simulation model with measured data of the normal gait. The differences in kinetic data, ground reaction force (GRF) and muscle force between the two models were analyzed to explore the different dynamic characteristic of hemiplegic gait and normal gait. Results The complex correlation coefficient between LifeMOD simulation results and measured data was 0.922, indicating that the established dynamic model was reasonable and effective. Hemiplegic patient with low tibialis anterior muscle strength led to ankle dorsiflexion inadequacy during initial ground period, and low gastrocnemius muscle could not achieve the promoting effect from ground during preswing period. Conclusions The strength weakness of tibialis anterior muscle and gastrocnemius are the main reasons for foot drooping and other hemiplegic gait characteristics. LifeMOD modeling and simulation can assist the diagnosis of abnormal muscle strength in hemiplegia patients.
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Objective To study the intrinsic relationship between hemiplegic gait features and abnormal muscle strength of lower limbs,so as to elucidate the muscle strength causes of hemiplegic gait and provide recommendations for its clinical treatment.Methods Models of hemiplegic gait and normal gait were established for simulation,and the validity of the models was verified by comparing the simulation model with measured data of the normal gait.The differences in kinetic data,ground reaction force (GRF) and muscle force between the two models were analyzed to explore the different dynamic characteristic of hemiplegic gait and normal gait.Results The complex correlation coefficient between LifeMOD simulation results and measured data was 0.922,indicating that the established dynamic model was reasonable and effective.Hemiplegic patient with low tibialis anterior muscle strength led to ankle dorsiflexion inadequacy during initial ground period,and low gastrocnemius muscle could not achieve the promoting effect from ground during preswing period.Conclusions The strength weakness of tibialis anterior muscle and gastrocnemius are the main reasons for foot drooping and other hemiplegic gait characteristics.LifeMOD modeling and simulation can assist the diagnosis of abnormal muscle strength in hemiplegia patients.
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Objective To establish qualitative and quantitative analysis methods for identification and determination of multi-constituent of Chanfukang granules (CFKG) using high performance liquid chromatography-time-of-flight mass spectrometry (HPLC-TOF-MS) and high performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-MS/MS),and a preliminary pharmacokinetic study was carried out.Methods An Agilent ZORBAX Eclipse Plus C1s column (100 mm × 3.0 mm,1.8 μm) and a Waters XBridge(R) BEH C18 column (150 mm × 4.6 mm,2.5 μm) were used with 0.1％ formic acid aqueous solution-acetonitrile as mobile phases by gradient elution.The compounds were detected by electrospray ion source in both positive and negative mode with multiple reaction monitoring (MRM) mode.SD rats were ig with CFKG (0.5,5 g/kg).Blood samples of 0.2 mL were taken from jugular vein at different time points and plasma components and contents were determined by HPLC-MS/MS.Results Twenty-four constituents were identified by HPLC-TOF-MS qualitative analysis and thirteen constituents were quantitatively detected by HPLC-MS/MS.The established HPLC-MS/MS method had a good separation,and all the legal verification met the requirements.The content of stachydrine,Leonurine,and astragaloside Ⅳ were high,which may be the main active ingredient The pharmacokinetic results showed that,only when the dose was 5 mg/kg,stachydrine and Leonurine can detected.Elimination of stachydrine was slow and elimination of Leonurine was fast.Conclusion A rapid and efficient method for studying the chemical constituents was established,which could provide reference for the quality control of Chanfukang granules.
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Objective To study and analyze the clinical efficacy of zoledronic acid in the treatment of osteoporotic intertrochanteric fracture in elderly patients. Methods 100 elderly patients with osteoporotic unstable intertrochanteric fracture treated in our hospital from February 2014 to July 2016 were selected and randomly divided into the control group and the experimental group. There were 50 patients in each group. The control group received routine treatment (calcitonin), and the experimental group was treated with zoledronic acid.The clinical indicators of the two groups were compared and analyzed. Results After the corresponding treatment, the VAS scores of the experimental group and the control group were (0.36±0.42) and (0.34±0.40), and there was no statistical significance. The excellent rate of hip joint function was 82% in the control group, and the excellent and good rate of hip joint function in the experimental group was 80%, which was not statistically significant. In addition, there were no statistically significant differences in fracture healing time and complications between the two groups. After 1 years, the hip bone mineral density and the lumbar vertebral bone mineral density were (0.78±0.05) g/cm2, and (0.85±0.06) g/cm2 were significantly higher than those in the control group,with statistical significance (P<0.05). Conclusion Zoledronic acid in the treatment of senile osteoporotic unstable intertrochanteric fracture clinical effect is remarkable,can increase bone mineral density in a large extent, the joint recovery had no effect, has clinical significance.
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Objective To study and analyze the clinical efficacy of zoledronic acid in the treatment of osteoporotic intertrochanteric fracture in elderly patients. Methods 100 elderly patients with osteoporotic unstable intertrochanteric fracture treated in our hospital from February 2014 to July 2016 were selected and randomly divided into the control group and the experimental group. There were 50 patients in each group. The control group received routine treatment (calcitonin), and the experimental group was treated with zoledronic acid.The clinical indicators of the two groups were compared and analyzed. Results After the corresponding treatment, the VAS scores of the experimental group and the control group were (0.36±0.42) and (0.34±0.40), and there was no statistical significance. The excellent rate of hip joint function was 82% in the control group, and the excellent and good rate of hip joint function in the experimental group was 80%, which was not statistically significant. In addition, there were no statistically significant differences in fracture healing time and complications between the two groups. After 1 years, the hip bone mineral density and the lumbar vertebral bone mineral density were (0.78±0.05) g/cm2, and (0.85±0.06) g/cm2 were significantly higher than those in the control group,with statistical significance (P<0.05). Conclusion Zoledronic acid in the treatment of senile osteoporotic unstable intertrochanteric fracture clinical effect is remarkable,can increase bone mineral density in a large extent, the joint recovery had no effect, has clinical significance.
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There is distinctive advantage of using male sterile lines to breed new cultivar and produce hybrids, when compared with general breeding method on yield and quality. In our previous work, near-isogenic lines (NILs) of male sterile and fertile Salvia miltiorrhiza have been obtained through continuous hybridization in many years. In this investigation, 378 primer combination were screened by using AFLP and BSA technique, in which 26 markers amplified from seven primers were found to tightly link to male sterile gene. Based on these markers, two linkage genetic maps were constructed. A 2 027,2 028 bp fragment was amplifed from NILs of fertile and sterile S. miltiorrhiza, respectively, using genome walking technique and previous E11/M4-208 marker as template. Four base mutations were found in intron when comparing both fragments. Among all different markers between NILs of male sterile and fertile S. miltiorrhiza, four was found to have 100% identities to chromosome 1, 3 and 5 of Arabidopsis, namely, E01/M09-418, E05/M13-308, E05/M04-750 and E01/M01-204. The E01/M09-418 marker was very close to male sterile gene of S. miltiorrhiza with distance of 2.1 cM, which also had 100% identities to male sterile gene MS2 in Arabidopsis. Both were distributed in chromosome 3 of Arabidopsis. The 2 028 bp fragment also had 100% identities to MS2 gene. Another E05/M04-750 marker that had 100% identities to chromosome 5 of Arabidopsis was found to have high identities to POP085-M05 gene of poplars and low affinity calcium antiporter CAX2 of Arabidopsis with very low E-value. The constructed genetic map and differential fragments with potential functions found in this study provide a solid foundation to lock male sterile genes in S. miltiorrhiza genome and to discover their functions.
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<p><b>BACKGROUND</b>Proteasome subunits (PSMB) and transporter associated with antigen processing (TAP) loci are located in the human leukocyte antigen (HLA) Class II region play important roles in immune response and protein degradation in neurodegenerative diseases. This study aimed to explore the association between single nucleotide polymorphisms (SNPs) of PSMB and TAP and Parkinson's disease (PD).</p><p><b>METHODS</b>A case-control study was conducted by genotyping SNPs in PSMB8, PSMB9, TAP1, and TAP2 genes in the Chinese population. Subjects included 542 sporadic patients with PD and 674 healthy controls. Nine identified SNPs in PSMB8, PSMB9, TAP1, and TAP2 were genotyped through SNaPshot testing.</p><p><b>RESULTS</b>The stratified analysis of rs17587 was specially performed on gender. Data revealed that female patients carry a higher frequency of rs17587-G/G versus (A/A + G/A) compared with controls. But there was no significant difference with respect to the genotypic frequencies of the SNPs in PSMB8, TAP1, and TAP2 loci in PD patients.</p><p><b>CONCLUSION</b>Chinese females carrying the rs17587-G/G genotype in PSMB9 may increase a higher risk for PD, but no linkage was found between other SNPs in HLA Class II region and PD.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , ATP Binding Cassette Transporter, Subfamily B, Member 2 , Genetics , ATP Binding Cassette Transporter, Subfamily B, Member 3 , Genetics , Antigen Presentation , Case-Control Studies , Cysteine Endopeptidases , Genetics , Parkinson Disease , Genetics , Allergy and Immunology , Polymorphism, Single Nucleotide , Proteasome Endopeptidase Complex , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effects of the combination of basic fibroblast growth factor (bFGF), transforming growth factor-Β1 (TGF-Β1), bone marrow mesenchymal stem cells (BMSCs), and temperature-responsive chitosan hydrogel (TCH) gel on the repair of degenerative intervertebral disc in rat models.</p><p><b>METHODS</b>Rat models of intervertebral disc degeneration were established by acupuncture. The degenerative effects were observed under magnetic resonance imaging (MRI). The BMSCs was cultured in vitro and then transfected by adenovirus with enhanced green fluorescent protein to make it carry the gene of enhanced green fluorescent protein,which functioned as fluorescence labeling. The SD rat models of intervertebral disc degeneration were divided into four groups: group A, treated with the combination of bFGF, TGF-Β1,BMSCs,and TCH gel; group B, treated with the combination of BMSCs and TCH gel;group C, treated with the combination of bFGF,TGF-Β1, and TCH gel;and group D, treated with PBS buffer solution. After the corresponding reagents were injected into the degenerative intervertebral discs of each group, the rats were cultivated for another four weeks and then the repair effects of the intervertebral discs were observed under MRI. Furthermore,the intervertebral discs of each group were taken out and observed by HE and Masson staining. The nucleus pulposus was aspirated and the expressions of aggrecan,collagen 2,Sox-9,and collagen I of nucleus pulposus of each group were tested by reverse transcription polymerase chain reaction and Western blot.</p><p><b>RESULTS</b>The transplanted BMSCs survived in the intervertebral disc and differentiated into nucleus pulposus-like cells. MRI showed that:the signal intensity of the nucleus pulposus of group A was much higher than that of the rest groups, the signal intensity of group B was higher than that of group C, and the signal intensity of group D was the lowest,in which the dura mater spinalis was in compression and the spinal cord changed in beaded shape. The differences of the Pfirrmann grading among the four groups had statistical significance (P<0.05). The results of the HE and Masson stains showed:the intervertebral disc of group A was well-structured,the quantity of nucleus pulposus cells was larger than that of the other three groups,and the boundary between the nucleus pulposus and the annulus fibrosus was clearly defined;the quantity of the nucleus pulposus cells of group B was larger than that of group C, and the broken annulus fibrosus was not observed in group B, while the broken annulus fibrosus could be observed in group C; and, the nucleus pulposus cells of group D were replaced by fibrous tissue. The results of the reverse transcription polymerase chain reaction and Western blot tests showed that,in terms of the expressions of aggrecan,collagen 2 and Sox-9,group A was the highest, followed by group B,group C,and group D (P<0.05); in terms of the expression of collagen 1,there was no obvious difference among these four groups (P>0.05).</p><p><b>CONCLUSIONS</b>The transplanted BMSCs can survive in the degenerative intervertebral disc and differentiate into nucleus pulposus-like cells. The combination of bFGF, TGF-Β1, BMSCs,and TCH gel has obvious repair effect on the degenerative intervertebral discs. The effect of the combination of BMSCs and TCH gel on transplantation therapy of the degenerative intervertebral discs is better than that of the combination of bFGF, TGF-Β1 and TCH gel but worse than that of the combination of bFGF, TGF-Β1, BMSCs, and TCH gel.</p>
Subject(s)
Animals , Rats , Bone Marrow Cells , Bone Marrow Transplantation , Cell Differentiation , Collagen , Disease Models, Animal , Fibroblast Growth Factor 2 , Hematopoietic Stem Cells , Intervertebral Disc , Intervertebral Disc Degeneration , Rats, Sprague-Dawley , Transforming Growth Factor beta1 , Wound HealingABSTRACT
<p><b>BACKGROUND</b>Nurr1 plays an essential role in the development, survival, and function maintenance of midbrain dopaminergic (DA) neurons, and it is a potential target for Parkinson's disease (PD). Nurr1 mRNA can be detected in peripheral blood mononuclear cells (PBMCs), but whether there is any association of altered Nurr1 expression in PBMC with the disease and DA drug treatments remains elusive. This study aimed to measure the Nurr1 mRNA level in PBMC and evaluate the effect of Nurr1 expression by DA agents in vivo and in vitro.</p><p><b>METHODS</b>The mRNA levels of Nurr1 in PBMC of four subgroups of 362 PD patients and 193 healthy controls (HCs) using real-time polymerase chain reaction were measured. The nonparametric Mann-Whitney U-test and Kruskal-Wallis test were performed to evaluate the differences between PD and HC, as well as the subgroups of PD. Multivariate linear regression analysis was used to evaluate the independent association of Nurr1 expression with Hoehn and Yahr scale, age, and drug treatments. Besides, the Nurr1 expression in cultured PBMC was measured to determine whether DA agonist pramipexole affects its mRNA level.</p><p><b>RESULTS</b>The relative Nurr1 mRNA levels in DA agonists treated subgroup were significant higher than those in recent-onset cases without any anti-PD treatments (de novo) (P < 0.001) and HC groups (P < 0.010), respectively. Furthermore, the increase in Nurr1 mRNA expression was seen in DA agonist and L-dopa group. Multivariate linear regression showed DA agonists, L-dopa, and DA agonists were independent predictors correlated with Nurr1 mRNA expression level in PBMC. In vitro, in the cultured PBMC treated with 10 μmol/L pramipexole, the Nurr1 mRNA levels were significantly increased by 99.61%, 71.75%, 73.16% in 2, 4, and 8 h, respectively (P < 0.001).</p><p><b>CONCLUSIONS</b>DA agonists can induce Nurr1 expression in PBMC, and such effect may contribute to DA agonists-mediated neuroprotection on DA neurons.</p>