Construction of a High-Throughput Mutation Library of Mycobacterium Rifampin Resistance Gene rpoB / 中山大学学报(医学科学版)

ObjectiveTo establish a mutation library of rifampicin resistance gene rpoB. MethodsThe ΔrpoB attB：：rpoB strain of Mycobacterium smegmatis （M. smegmatis） be constructed by homologous recombination and L5 attB phage integration site exchange. Based on the L5 attB plasmid exchange system and resistance selection medium， 48 clones are selected to verify plasmid replacement efficiency. Degenerate primers are designed every 3 bases in the rifampicin resistance determining region （RRDR）， and a full-coverage mutation library of 81 bases in RRDR region is obtained by PCR amplification. The library fragments are seamlessly cloned into the vector and transformed into Escherichia coli （E. coli）to form an E. coli mutation library. Based on the principle of plasmid exchange， the mutant plasmid library is transformed into the M. smegmatis strain ΔrpoB attB：：rpoB， and the original L5 attB site plasmid is replaced to form the M. smegmatis mutant library. The genotype of the library are determined by genome extraction， library construction and high-throughput sequencing. ResultsCompared with the wild-type rpoB gene （5 600 bp）， the amplified fragment of the rpoB knockout strain is 2 200 bp， which proved that the ΔrpoB attB：：rpoB conditional knockout strain of M. smegmatis is successfully constructed. The success rate of plasmid replacement is 100%. There were 540 kinds of single amino acid mutations in both E. coli library and M. smegmatis library， 5 301 kinds of multi-point mutations in E. coli library， and 853 kinds of multi-point mutations in M. smegmatis library. The correlation coefficient between E. coli library and M. smegmatis library is 0.84. ConclusionsWe have developed a strategy to construct a library of mutants targeting the essential mycobacterial gene rpoB， and successfully established a mutant library of rifampicin resistance gene rpoB.

Application of 3D printing percutaneous guide plate in closed reduction and cannulated screw internal fixation of femoral neck fracture / 中国骨伤

OBJECTIVE@#To investigate the application of 3D printing percutaneous surgical guide plate in closed reduction and cannulated screw internal fixation of femoral neck fracture.@*METHODS@#The clinical data of 12 patients with femoral neck fracture from March 2019 to March 2022 were retrospectively analyzed. Patients were divided into observation group and control group according to different operation plans, with 6 cases in each group. The observation group received percutaneous operation guide plate assisted closed reduction and hollow screw internal fixation, while the control group received closed reduction and hollow compression screw internal fixation. The operation time, intraoperative blood loss, fluoroscopy times, and Kirschner needle puncture times were compared between two groups. The location of screws were recordedon postoperative X-ray films, follow-up time, time of complete fracture healing, Harris score of hip joint and the incidence of complications were recorded on postoperative X-ray films.@*RESULTS@#The operation time of observation group (32.17±6.18) min was shorter than that of control group (53.83±7.31) min (P<0.05). The amount of intraoperative bleeding in the observation group (18.33±2.94) ml was less than that in the control group (38.17±5.56) ml(P<0.05). The times of fluoroscopy in the observation group (7.50±1.05) were less than those in the control group (21.00±4.82) (P<0.05). The number of Kirschner needle punctures (8.00±0.63) in observation group was less than that in control group (32.67±3.08) (P<0.05). The follow-up time was(12.88±0.74) months in observation group and (12.83±0.72) months in control group, there was no significant difference between two groups (P>0.05). One year after operation, Harris score of hip joint in the observation group was(82.00±4.52) points, while that in the control group was(81.00±3.41) points, there was no significant difference between two groups(P>0.05). The time of complete fracture healing in the observation group was (7.50±1.05) months, while that in the control group was (7.67±1.21) months, there was no significant difference between two groups(P>0.05). The parallelism of the screws in the observation group was (0.50±0.11) ° and (0.76±0.15) °, which were lower than that in the control group (1.57±0.31) ° and (1.87±0.21) ° (P<0.05). The screw distribution area ratio (0.13±0.02) cm2 in the observation group was higher than that in the control group (0.08±0.01) cm2 (P<0.05). No complications such as necrosis of femoral head, nonunion of fracture, shortening of femoral neck and withdrawal of internal fixation occurred in both groups.@*CONCLUSION@#The application of 3D printing percutaneous surgical guide plate improves the accuracy and safety of closed reduction and cannulated screw internal fixation for femoral neck fracture. It has the advantages of minimally invasive, reducing radiation exposure, fast and accurate, shortening the operation time and reducing intraoperative bleeding.

Risk factors of low cardiac output syndrome after cardiac valvular surgery in elderly patients with valvular disease complicated with giant left ventricle / 中华心血管病杂志

Objective: To explore the risk factors of low cardiac output syndrome (LCOS) after cardiac valvular surgery in elderly patients with valvular disease complicated with giant left ventricle. Methods: This was a retrospective study. The clinical data of patients over 60 years old with giant left ventricle who underwent cardiac valvular surgery in Henan Provincial People's Hospital (Fuwai Central China Cardiovascular Hospital) from January 2016 to January 2020 were collected in this study. Patients were divided into LCOS group and non-LCOS group. The clinical data, preoperative echocardiographic results and surgical data of all patients were collected. Taking LCOS as dependent variable and statistically significant variables in univariate analysis as independent variable, multivariate logistic regression equation was constructed to identify the risk factors of LCOS after cardiac valvular surgery in elderly patients with valvular disease complicated with giant left ventricle. On the basis of logistic regression, the risk factors of continuous variables were put into the regression model for trend test. Results: A total of 112 patients were included, among whom 76 patients were male, the mean age was (65.3±3.8) years. There were 21 cases in LCOS group and 91 cases in non LCOS group. Univariate analysis showed that age≥70 years, preoperative NYHA cardiac function class Ⅳ, preoperative renal dysfunction, preoperative cerebrovascular disease, preoperative LVEF<40%, blood loss/total blood volume>20%, cardiopulmonary bypass (CPB) time>130 minutes and aortic cross-clamp time>90 minutes all had statistically significant differences between the two groups (all P<0.05). Multivariate logistic regression analysis showed that age≥70 years (OR=5.067, 95%CI 1.320-19.456, P=0.018), preoperative NYHA cardiac function class Ⅳ (OR=3.100, 95%CI 1.026-9.368, P=0.045), renal dysfunction (OR=3.627, 95%CI 1.018-12.926, P=0.047), CPB time>130 minutes (OR=4.539, 95%CI 1.483-13.887, P=0.008) were the independent risk factors of LCOS after cardiac valvular surgery in elderly patients with giant left ventricle. Risk of LCOS was significantly higher in patients aged from 65 to 70 years (OR=1.784, 95%CI 0.581-5.476) and aged 70 years and above (OR=4.400, 95%CI 1.171-16.531) than in patients aged from 60 to 65 years. The trend test results showed that the risk of LCOS increased significantly in proportion with the increase of age (P for trend=0.024). Risk of LCOS was significantly higher in patients with CPB time between 90 and 110 minutes (OR=1.917, 95%CI 0.356-10.322), 110 and 130 minutes (OR=1.437, 95%CI 0.114-18.076) and 130 minutes and above (OR=5.750, 95%CI 1.158-28.551) than in patients with CPB time ≤ 90 minutes (P for trend=0.009). Conclusions: The risk factors of LCOS after cardiac valvular surgery are age≥70 years, preoperative NYHA cardiac function class Ⅳ, renal dysfunction, CPB time>130 minutes in elderly patients with giant left ventricle.

Clinical efficacy of two different tricuspid annuloplasty techniques in left cardiac valve surgery

Objective:To investigate the mid-and long-term clinical effects of tricuspid valvuloplasty with the implantation of an artificial plastic ring.Methods:Data of 677 patients who had functional tricuspid regurgitation and left cardiac valve disease and underwent tricuspid valvuloplasty and left cardiac valve surgery were retrospectively. Among these patients, 353 underwent simple suture annuloplasty (group A) while the rest 324 patients underwent artificial plastic ring annuloplasty (group B). The two-year and more-than-two-year clinical and ultrasonocardiograph (UCG) follow-up data of the two groups were obtained and compared.Results:A total of 600 patients (88.6%) completed the long-term follow-up (more than two years). The two-year follow-up showed no significant difference in the incidence of mild tricuspid regurgitation between the two groups (82.2% vs. 92.7%, P=0.37). However, there were significantly more cases that developed into moderate to severe tricuspid regurgitation in group A than in group B (17.8% vs. 7.3%, P=0.031). The long-term follow-up revealed that the recurrence rate of tricuspid regurgitation in group B was significantly lower than that in group A (11.0% vs. 25.0%, P=0.029), and the ratio of cases developing into moderate to severe tricuspid regurgitation in group A was significantly higher than that in group B (28.9% vs. 9.9%, P=0.007). The comparison between the two intra-group time segments showed that the development of tricuspid regurgitation in group A was significantly increased (28.9% vs. 17.8%, P=0.022), but in group B it was relatively stable (9.9% vs. 7.3%, P=0.52).Conclusions:Artificial ring annuloplasty is associated with significantly less tricuspid regurgitation than simple suture annuloplasty.

Clinical features and prognosis of patients with acute ST-segment elevation myocardial infarction comorbid with diabetes mellitus / 中国介入心脏病学杂志

Objective To investigate the clinical features of patients with acute ST-segment elevation myocardial infarction (STEMI) comorbid with diabetes mellitus (DM) and to analyze the prognosis within 12 months after primary percutaneous coronary intervention (pre-PCI). Methods A total of 375 STEMI patients were divided into the diabetes group (n=140) and the normal blood glucose group(n=235) according to whether they met the diagnostic criteria of DH. The clinical data,characteristics of coronary artery lesions,type of stent implant,rate of coronary slow flow or no-reflow after pre-PCI, and the prognosis within 12 months after PCI of the two groups were investigated.Results Patient in the diabetes group presented with higher mean age ,higher comorbid rates of hypertension , hyperlipidemia and heart function of Killip class Ш and above than patients in the normal blood glucose group (all P<0.05). patients in the diabetes group had higher rates of slow reflow /no-reflow after PCI(12.9% vs.5.5%,P=0.013),higher percentages of 3-ressel disease(40.7% vs. 28.9%,P=0.019)and lef t main lesions(13.6% vs. 7.2%,P=0.044). The in-hospital mortality rates(6.4% vs.1.7%,P=0.020),revascularization rates within 12 months(7.9% vs.0.9%,P=0.001)and incidence of heart failure(7.9% vs. 2.6%,P=0.017)were all higher in the diabetes group. Conclusions STEMI patients comorbid with DM were relatively older, had higher comorbidities of hypertension,hyperlipidemia, three-vessel disease, left main coronary lesions and higher mortality during hospitalization. No significant increase in cardiac death and recurrent myocardial infarction were deserved during the follow-up period. These patients may benefit more from early intervention.

Related factors for dysphagia after single-level anterior cervical descectomy and fusion / 中国组织工程研究

BACKGROUND: Patients with anterior cervical discectomy and fusion have a high incidence of dysphagia, which may be associated with a variety of factors. The specific mechanism remains unclear. OBJECTIVE: To explore the related factors of dysphagia after single-level anterior cervical discectomy and fusion. METHODS: We retrospectively analyzed patients with cervical degenerative disc disease receiving single-level anterior cervical discectomy and fusion in First Affiliated Hospital of Soochow University from January 2011 to January 2015. During hospitalization, basic patient data and surgery-related data were recorded, including operation time, intraoperative blood loss, surgical segment, internal fixation device and the use of recombinant human bone morphogenetic protein-2. The cervical alignment and prevertebral soft tissue swelling were measured preoperatively and 3 days postoperatively. At 1, 3, 6, 12, and 24 months postoperatively, the Bazaz swallowing function scoring system was used to assess the swallowing of the patients. RESULTS AND CONCLUSION: A total of 262 patients undergoing single-level anterior cervical discectomy and fusion were involved. The incidence of dysphagia at 1, 3, 6, 12, and 24 months postoperatively was 35.9%, 22.9%, 15.6%, 11.5% and 9.2% respectively. Univariate analysis showed that gender, operation time and course length were associated with postoperative dysphagia. Logistic multivariate regression analysis showed that the duration of operation (≥ 3 hours), female and course length (≥ 8 months) were risk factors for dysphagia after anterior cervical descectomy and fusion. Operation time and female may be associated with early and middle dysphagia postoperatively, and the course length may be associated with chronic dysphagia. Prevertebral soft tissue swelling and other factors are not related to dysphagia after single-level anterior cervical discectomy and fusion. Risk factors for dysphagia after multi-level fusion should be further studied.

A study on the degradation of anti-influenza drug GZ830 / 国际药学研究杂志

Objective To establish an HPLC method for the determination of anti-influenza drug GZ830 and its related substances, and investigate the degradation of GZ830. Methods The degradation of GZ830 under the conditions with different pH and temperatures was investigated with HPLC method to explore the degradation principle. Results The HPLC method for the determination of GZ830 was established. The linear relationship between the drug and the peak area was good in the concentration range of 160-1200 μg/ml (r=0.9998). GZ830 was easily degraded under alkaline conditions and could not tolerate high temperatures. The degradation of GZ80 did not occur when its aqueous solution was kept at 40℃ for 6 h. However, after heating at 80℃ for 6 h, about 2.4％GZ80 was degraded and the degradation rate reached about 7％ after kept at 115℃ for 30 min. Conclusion Under the optimized chromatographic conditions, GZ830 was well separated from the degraded products, and the HPLC method possessed high sensitivity and good specificity for the determination of GZ80, and the degradation of GZ80 was different under different conditions.

Clinical efficacy of two different tricuspid annuloplasty techniques in left cardiac valve surgery

Objective: To investigate the mid-and long-term clinical effects of tricuspid valvuloplasty with the implantation of an artificial plastic ring. Methods: Data of 677 patients who had functional tricuspid regurgitation and left cardiac valve disease and underwent tricuspid valvuloplasty and left cardiac valve surgery were retrospectively. Among these patients, 353 underwent simple suture annuloplasty (group A) while the rest 324 patients underwent artificial plastic ring annuloplasty (group B). The two-year and more-than-two-year clinical and ultrasonocardiograph (UCG) follow-up data of the two groups were obtained and compared. Results: A total of 600 patients (88.6%) completed the long-term follow-up (more than two years). The two-year follow-up showed no significant difference in the incidence of mild tricuspid regurgitation between the two groups (82.2% vs. 92.7%, P=0.37). However, there were significantly more cases that developed into moderate to severe tricuspid regurgitation in group A than in group B (17.8% vs. 7.3%, P=0.031). The long-term follow-up revealed that the recurrence rate of tricuspid regurgitation in group B was significantly lower than that in group A (11.0% vs. 25.0%, P=0.029), and the ratio of cases developing into moderate to severe tricuspid regurgitation in group A was significantly higher than that in group B (28.9% vs. 9.9%, P=0.007). The comparison between the two intra-group time segments showed that the development of tricuspid regurgitation in group A was significantly increased (28.9% vs. 17.8%, P=0.022), but in group B it was relatively stable (9.9% vs. 7.3%, P=0.52). Conclusions: Artificial ring annuloplasty is associated with significantly less tricuspid regurgitation than simple suture annuloplasty.

Effect of FSD-C10 on modulation of inflammatory microenvironment in an Alzheimer disease double transgenic mouse model / 中国病理生理杂志

AIM:To explore the therapeutic effect of a novel Rho kinase inhibitor FSD-C10 onβ-amyloid pro-tein precursor (APP)/presenilin-1 (PS1) double transgenic mice.METHODS: The transgenic mice overexpressing hu-man APP with the Swedish mutation (695) and human PS1 with ΔE9 mutation at the age of 8 months were used in this study.The mice were randomly divided into model group and FSD-C10 intervention group, and wild-type mice at the same age served as normal controls .The mice in FSD-C10 intervention group were treated with FSD-C10 (25 mg· kg-1 · d-1 ) for 2 months by intraperitoneal injection .The mice in model group and the wild-type mice were injected with saline in the similar manner.Morris water maze (MWM) test was applied to examine the capacity of learning and memory .The Aβ1-42 deposition, Tau protein phosphorylation , and the expression of β-site APP-cleaving enzyme ( BACE) as well as inflammato-ry molecules, such as TLR-4 and NF-Κb, and M1/M2 microglial markers, such as Inos and Arg-1, were determined by the methods of immunohistochemistry and Western blot .RESULTS: Compared with model group , FSD-C10 significantly improved the learning and memory abilities of APP/PS1 double transgenic mice , accompanied by reduced Aβ1-42 deposi-tion, Tau protein phosphorylation and BACE expression in the hippocampus .The intervention of FSD-C10 decreased the protein levels of TLR-4 and p-NF-Κb, reduced the expression of Inos and increased the expression of Arg-1 in the brain tissues.CONCLUSION:The novel Rho kinase inhibitor FSD-C10 improves the capacity of spatial learning and memory in APP/PS1 double transgenic mice , which may be related to the inhibition of TLRs/NF-Κb signaling pathway , the reduction of the secretion of inflammatory molecules and the polarization of anti-inflammatory M2 microglia, thus improving the in-flammatory microenvironment of the brain in APP/PS1 double transgenic mice .

Effect of FSD-C10 on modulation of inflammatory microenvironment in an Alzheimer disease double transgenic mouse model / 中国病理生理杂志

AIM:To explore the therapeutic effect of a novel Rho kinase inhibitor FSD-C10 onβ-amyloid pro-tein precursor (APP)/presenilin-1 (PS1) double transgenic mice.METHODS: The transgenic mice overexpressing hu-man APP with the Swedish mutation (695) and human PS1 with ΔE9 mutation at the age of 8 months were used in this study.The mice were randomly divided into model group and FSD-C10 intervention group, and wild-type mice at the same age served as normal controls .The mice in FSD-C10 intervention group were treated with FSD-C10 (25 mg· kg-1 · d-1 ) for 2 months by intraperitoneal injection .The mice in model group and the wild-type mice were injected with saline in the similar manner.Morris water maze (MWM) test was applied to examine the capacity of learning and memory .The Aβ1-42 deposition, Tau protein phosphorylation , and the expression of β-site APP-cleaving enzyme ( BACE) as well as inflammato-ry molecules, such as TLR-4 and NF-Κb, and M1/M2 microglial markers, such as Inos and Arg-1, were determined by the methods of immunohistochemistry and Western blot .RESULTS: Compared with model group , FSD-C10 significantly improved the learning and memory abilities of APP/PS1 double transgenic mice , accompanied by reduced Aβ1-42 deposi-tion, Tau protein phosphorylation and BACE expression in the hippocampus .The intervention of FSD-C10 decreased the protein levels of TLR-4 and p-NF-Κb, reduced the expression of Inos and increased the expression of Arg-1 in the brain tissues.CONCLUSION:The novel Rho kinase inhibitor FSD-C10 improves the capacity of spatial learning and memory in APP/PS1 double transgenic mice , which may be related to the inhibition of TLRs/NF-Κb signaling pathway , the reduction of the secretion of inflammatory molecules and the polarization of anti-inflammatory M2 microglia, thus improving the in-flammatory microenvironment of the brain in APP/PS1 double transgenic mice .

Proton pump inhibitors and hypomagnesemia / 国际药学研究杂志

Proton pump inhibitors (PPI) are currently the most effective drugs for the treatment of acid related diseases. They are safe in short-term use, but can cause a series of security problems after long-term use. Hypomagnesemia caused by PPI is an important adverse drug reaction with reversible effect and has the following features: it only happens after long-term treatment with the extremely low occurrence rate; the proportion is higher in the elderly and female patients and usually independent of PPI dosage; magnesium supplementation is not effective for hypomagnesemia; the risk of hypomagnesemia increases when PPI is combined with diuretics, nephrotoxic drugs and drugs of reducing serum magnesium; H2-receptor antagonist is the best alternative of PPI. In this paper, we review hypomagnesemia induced by PPI.

Drug-resistant and molecular characteristic of Klebsiella pneumoniae isolated from nosocomial and animal origins / 中国人兽共患病学报

We aimed to detect the drug resistant and molecular characteristic of Klebsiella Pneumoniae (KP) isolated from different hosts origin,and to investigate the infection possibility between animals and humans.A total of 98 KP strains were collected in Henan Province from March 2013 to December 2014.Drug-resistance to 15 antibiotics was detected by K-B diffu sion methods.The phenotype of produce mucus was determined by stringing test.Seven drug-resistant gene and 2 virulence gene were amplified by PCR technique.Molecular types were analysis by MLST.The resistant rate of KP isolated from noso comial was higher than those isolated from animals.Among animal strains,the resistant rate of KP isolated from chickens and pigs were higher than that from rabbits and dogs.The multidrug resistant (MDR) of nosocomial isolates were the highest (74.19％).There were 18 STs among 98 KP strains.The main prevalent types were ST37 in chicken,ST258 in pigs,ST60 in rabbits,ST11 in dogs and nosocomial respectively.ST11 was common epidemic types among nosocomial,dogs and pigs.ST235 was common molecular types among chicken and human.ST258 existed in both nosocomial and pigs.The rmpA gene and magA gene were detected in ST11,ST235 and ST 258,producing higher mucus.The blaKPC gene was 54.84％ in nosocomial strains,but not been detected in dog and rabbit strains.The distribution of ESBLs gene in nosocomial was higher than those in animals,but qnrA gene and qnrB gene were higher in chicken than in human.The multidrug resistant gene was the highest distribution in ST11,ST258 and ST235.There were some differences in phenotype of drug-resistance,producing mucus and molecular characteristics,but ST11,ST258 and ST235 were common types of KP isolates from different host origins.

"Time-angle measurement" reduction evaluation technique and clinical evaluation of proximal humerus fracture / 北京大学学报(医学版)

Objective:To evaluate the functional outcome and the complication rate after open reduction and internal fixation of proximal humeral fractures with a "time-angle measurement" reduction evaluation technique,to assess the acceptance of the fracture reduction and to estimate the result of the surgery.Methods:Forty-six patients [mean age:(66.2 ± 14.9) years] with an acute proximal humeral fracture were managed with open reduction and internal fixation with this reduction evaluation technique from January 2012 to December 2013.According to the Neer classification,there were 6 two-part fractures,25 three-part fractures and 15 four-part fractures.The functional outcome was evaluated for each patient using the Constant-Murley score;radiographic evaluation was also conducted and complications were recorded.The postoperative shoulder function recovery and imaging findings were followed up to evaluate the guiding significance of this reduction evaluation technique in the clinical treatment of this kind of fracture.Results:In the study,46 patients had been followed up for 13-36 months,and the average follow-up time was (23.5 ± 7.3) months.All the patients achieved fracture healing 3 months after operation.The average head-shaft angle was (124 ± 3.5) degrees.According to the Constant scoring system,29 patients (63％) had excellent,14 patients (30％) had good,and 3 patients (7％) had poor results.The most common complications were pain (7/65) and restricted movement of the shoulder (5/ 46).There were no cases of screw penetration,necrosis of humeral head,deep tissue infection,nonunion of fracture and axillary nerve injury after operation.Conclusion:For appropriate cases of displaced proxinal humeral fractures,surgical treatment with application of "time-angle measurement" reduction evaluation technique that was introduced in the present study can lead to a good functional outcome,and the technique of reduction assessment should be regarded as a reasonable reference standard in the treatment of displaced proximal humerus fracture.

Neuroprotective effect of fasudil combined with bone marrow-derived neural stem cells on mice with experimental autoimmune encephalomyelitis / 中国病理生理杂志

AIM: To explore the neuroprotective effect of fasudil combined with bone marrow -derived neural stem cells ( BM-NSCs) on the mice with experimental autoimmune encephalomyelitis ( EAE).METHODS: Female C57BL/6 mice (8~10 weeks old, n=32) were immunized with myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) to establish chronic EAE model .The mice were randomly divided into control ( ddH2 O ) group, fasudil group , BM-NSCs group , and fasudil+BM-NSCs group .The clinical score and body weight were recorded every other day .The expression of neurotrophic factors was determined by immunofluorescence staining .RESULTS:In comparison with ddH2O group, fasud-il combined with BM-NSCs delayed onset and ameliorated severity of EAE .The numbers of brain-derived neurotrophic fac-tor, glial cell-derived neurotrophic factor , nerve growth factor , neurotrophin-3 and ciliary neurotrophic factor positive cells in fasudil group, BM-NSCs group and fasudil +BM-NSCs group were all increased in various extents .In particularly, the expression of these neurotrophic factors in fasudil +BM-NSCs group was significantly higher than that in the mice treated with fasudil or BM-NSCs alone (P<0.01).CONCLUSION:Fasudil combined with BM-NSCs promotes the expression of neurotrophic factors and improves microenvironment of central nervous system , thus playing a positive role in neural restora-tion and regeneration through a synergistic and superimposed effect .

Knockout gata4 gene and establish ment of a zebrafish model of congenital heart disease by TALEN / 中国比较医学杂志

Objective To establish a gata4 gene knockout zebrafish model of congenital heart disease, and construct transcription activator-like effector nuclease ( TALEN) vectors targeting gata4 gene.Method We construct TALEN vectors targeting zabrafish gata4 gene using unit assembly method and the in vitro-transcribed TALEN mRNAs were microinjected into one-cell stage zebrafish embryos.The efficiency of TALEN was identified by injected embryos, and mutations of zebrafish were screened and confirmed the different types through PCR and enzyme digestion.Results We successfully constructed correct targeting vectors by enzyme digestion and sequencing, and the gene knockout efficiency was 35.18%.We screened the mutant zebrafish and confirmed different types of gata4 gene mutations.Conclusions A gata4 knockout zebrafish model is successfully established, it can provide a good animal model for further research of congenital heart diseases.

Individualized internal fixation plate for tibial fracture by digital customization and template for minimally invasive surgery / 医用生物力学

Objective To investigate the process and method of making an individualized limited contact dynamic compression plate (LC-DCP) and template for fixing tibial fracture, as well as its preliminary effect in clinic. Methods The custom-made tibia LC-DCP and the template used in minimally invasive percutaneous plate osteosynthesis (MIPPO) surgery were developed by 3D reconstruction of CT images, computer-aided design/manufacturing (CAD/CAM) and computer numerical control (CNC) technology, and then applied in 6 patients with tibial fractures. Results The individualized tibia LC-DCP was successfully used in clinic. The average 20-month follow-up showed that all fractures were healed in 6 patients, with 5 reaching excellent and 1 reaching good standard, according to criteria of Johner-Wruhs. Conclusions The production process of such individualized plate is simple and practicable, and has the advantages of simple operation, less injury, satisfactory reduction and fixation, high healing rate, less complexity when treating tibial fracture in MIPPO surgery. Therefore, it is worth of clinical application.

Effects of oscillatory flow on shear stress distributions in perfusion bioreactor for bone tissue engineering / 医用生物力学

Objective To study the effects of oscillatory flow, as well as pore size and porosity of the 3D scaffold on distributions of flow rate and shear stress in perfusion bioreactor, and to propose optimization methods for preparing the 3D decellularized bone scaffold and perfusion bioreactor based on theoretical results. Methods Based on the previously established 3D decelluarized scaffold and perfusion bioreactor for bone tissue engineering in the laboratory, the decelluarized scaffold was simplified as an isotropic porous media. The velocity and shear stress distributions in the bioreactor were further simulated theoretically. Results Under the oscillatory flow, the Darcy shear stress and velocity in the 3D porous scaffold presented a consistently regular pattern. Compared with the unidirectional flow, the difference of velocity and Darcy shear stress decreased at different radius, which could contribute to the homogeneous 3D culture of seed cells in bone tissue engineering. Increasing the inlet perfusion velocity could improve the average Darcy shear stress. Increasing the pore diameter or porosity of the scaffold had no obvious effects on the peak value of velocity, but sharply reduced the average Darcy shear stress. Increasing inlet oscillation frequency could decrease the peak value of velocity and obviously decrease the difference of velocity at different radius. Conclusions Appropriate oscillatory flow was beneficial for generating required shear stress for stem cells in bone tissue engineering. The research findings in this study are expected to provide theoretical guidance to optimize the 3D culture method of seed cells for bone tissue engineering.

A case-control study on the relationship of drinking water from farmland edge and digestive tract cancers / 预防医学

Objective To explore the relationship between drinking water from farmland edge and digestive tract cancers in rural areas.Methods A total of 180 cases of digestive tract cancers from Longquan city,Jinyun county and Qingtian county were selected and 360 controls with the same age group and sex were enrolled.1∶2 matched design of case-control study was used.The data was analyzed by Wilcoxon matched -pairs signed -ranks test.Results Overall,individuals who drank water from farmland edge were more likely to have digestive tract cancer among 3 areas (OR=4.005 ,95%CI:2.228-7.199).In Qingtian county the risk of digestive tract cancers in individuals who drank water from farmland edge was 5.375 times of control group population (95%CI:2.287 -12.635 ),But there was not statistically significant in Longquan city and Jinyun county (P >0.05 ).For gastric cancer,the proportion of drinking farmland edge water was significantly higher in cases than that in control group (OR=4.875,95%CI:2.007-11.839),while the difference was not statistically significant for liver cancer,colorectal cancer and esophageal cancer (P>0.05 ).In Qingtian county ,for gastric cancer,the proportion of drinking water from farmland edge was 5.750 times of control group population (95%CI:1.683-19.649).But there was not statistically significant in other cancers(P>0.05).Conclusion Drinking water near farmland edge may be one of the risk factors of gastric cancer in rural areas.

Research of zebrafish quality standardization / 中国实验动物学报

At present, zebrafish has played an increasingly important role in models for human development and diseases and several areas of life sciences.As a newly laboratory animal resource, standardization research has become the technical bottleneck to be solved and an inevitable trend.In this review, we summarized the research history and character-istics of zebrafish and the status of quality standardization.We also discussed the main problem facing by the standardiza-tion research of zebrafish as a newly laboratory animal.We hope that the data can provide useful reference for the develop-ment of zebrafish quality standardization research.

Effects of triamcinolone acetonide and Ketorolac tromethamine on the expressions of aquaporin-4 and vascular endothelial growth factor in hypoxic Müller cell / 中华实验眼科杂志

Background Intravitreal injection with triamcinolone acetonide (TA) may cause complications,including increase of intraoculapressure (IOP),cataracand endophthalmitis.Ketorolatromethamine (Ketorolac) inew,lesadverse reactionof non-steroidal anti-inflammatory drug.The action mechanism of Ketorolaisimilato TA.Therefore,Ketorolamay be completely opartly replace Tin the treatmenof retinal edema.Objective The purpose of thistudy wato investigate the effectof Tand Ketorolaon the expressionof aquaporin-4 (AQP4) and vasculaendothelial growth facto(VEGF) in hypoxiretinal Müllecellin vitro and to explore the mechanism of treating retinal edemwith Tand Ketorolac.MethodThe propose of research and use of the animalwere approved by Animal ExperimenResearch Review Committee of Hubei University of Medicine.Twenty eyeof New Zealand albino rabbitwere extracted and the retinal tissue waisolated.The Müllecellwere cultured and passaged using the enzymatidigestion method and Müllecellwere identified using glial fibrillary acidiprotein (GFAP),vimentin and α-smooth muscle actin (α-SMA) by immunofluorescence staining.The hypoxicell modelwere established by culturing the cellin DMEM with 500 μmol/L CoCl2 fo0,6,12,24 hours.The cellof hypoxifo24 hourwere divided into normal control group,hypoxicontrol group,hypoxia+50,100,200 mg/L To50,100,200 mg/L Ketorolagroups.Corresponding drugwere added into the medium in the differengroups.The expressionof AQP4 mRNand VEGF mRNin Müllecellwere detected by semi-quantitative reverse transcription PC(RT-PCR).ResultThe cellgrew well and reached 80％ confluence with the irregulashape and ovoid nuclei 14-15 dayaftecultured.More than 95％ primary cellshowed positive reaction to GFAP,vimentin and α-SMA.The expressing levelof AQP4 mRNand VEGF mRNin Müllecell(values) were significantly differenin varioutime point(AQP4 mRNA:F=18.70,P＜0.01 ; VEGF mRNA:F =53.20,P＜0.01),and those of 6,12 and 24 houraftecultured with CoCl2were increased than those withouCoCl2 (P＜0.05).The expressing levelof AQP4 mRNand VEGF mRNin Müllecell(values) were significandifferenamong the normal control group,hypoxicontrol group,hypoxia+50,100,200 mg/L ToKetorolagroup(AQP4 mRNA:F =27.98,P ＜ 0.01 ; VEGF mRNA:F =10.03,P ＜0.01).Compared with the hypoxicontrol group,the expressing levelof AQP4 mRNand VEGF mRNin the Müllecellwere declined in the hypoxia+ 100,200 mg/L Tgroup and the hypoxia+100,200 mg/L Ketorolagroup (P＜0.05).The expressing levelof AQP4 mRNand VEGF mRNwere found statistically insignificandifference between hypoxia+ 100 mg/L Tgroup and hypoxia+ 100 mg/L Ketorolagroup,obetween hypoxia+ 200 mg/L Tgroup and hypoxi+200 mg/L Ketorolagroup (P＞ 0.05).ConclusionTand Ketorolacan downregulate the expressionof AQP4 and VEGF in Müllecellundehypoxiconditions,inferring thathey have similamechanism in the impacon AQP4 function in retinal edematoueye.