Objective To observe the expression of neuron specific enolase (NEC) to evaluate the neuroprotective effect of a cell-penetrating Bax-inhibiting peptide (BIP) on neonatal rats with hypoxic-ischemic brain damage (HIBD). Methods Wi-star rats (7-day old) were randomly divided into Sham group, BIP group and HIBD group. After modeling HIBD, the histologi-cal (HE staining) and immunohistochemistry methods were used to determine the apoptotic pathological changes and the NSE expression levels in the brain at different time points. Results Compared to the Sham group, the rats of HIBD group showed significant apoptotic pathological changes. The histological changes and the brain damages were improved significantly in BIP group at each sampling point. The number of NSE-positive cells was significantly decreased in HIBD and BIP groups over time (P<0.05). The number of NSE-positive cells had significant difference among different groups at 48 h, 96 h and 7 d after opera-tion (F=45.35-81.66, P<0.01). The number of NSE-positive cells in the HIBD group was smaller than that of the Sham group and BIP group 48 h after operation (P<0.05). The number of NSE-positive cells in the BIP group was smaller than that of the Sham group 96 h after operation (P<0. 05). Conclusions BIP can decrease the apoptosis of cortex nerve cells in 7-day old HIBD rat model, and may have neuroprotective effect on the early stage of HIBD.

Recurrent intracerebral hemorrhage is a very serious cerebrovascular disease.Its incidence has shown a clear upward trend, and both disability and mortality have increased significantly compared to the first intracerebral hemorrhage. This article reviews the clinical classification, characteristics, risk factors, pathogenesis, and prognosis of recurrent intracerebral hemorrhage.

ObjectiveToevaluatethecorrelationbetweenfluoroquinoloneresistanceinNeisseriagonor-rhoeaeandmutationsingyrAandparCgenes.Methods①Thesusceptibilities58clinicalisolatesofN.gonorrhoeaeto5fluoroquinolonesweretestedbydiscdiffusionmethod.②Theminimuminhibitoryconcentration(MIC)ofciprofloxacinwasdeterminedbyE-test.③Thefragmentsincludingthequinoloneresistance-determiningregion(QRDR)wereamplifiedbyPCRingyrAgeneof18strains,andparCgeneof8strains,andtheirrelativefragmentsweredirectlysequenced.Results①Thenumbersofstrainssimultaneouslysensitive,intermediateandresistanttociprofloxacin,ofloxacin,lomefloxacin,fleroxacinandenoxacinwere2,4and39,respectively.②TherangeofciprofloxacinMICwas0.004～12.0?g/mLin58strains.Thenumbersofstrainssensitive,intermediateandresistanttociprofloracinwere2,17and39,respectively.③ThestrainswithciprofloxacinMICfrom0.004～0.016?g/mLhadnomutationingyrAandparCgenes.ThestrainswithMICfrom0.064to0.094?g/mLcarriedasinglepointmutationingyrAgene,whilethestrainswithMIC≥0.25?g/mLcontainedtwomutationsingyrAgene.Inaddition,thestrainswithMIC≤0.25?g/mLhadnomutationinparCgeneandthestrainswithMIC≥1.0?g/mLexhibitedasinglepointmutationinparCgeneandtwomutationsingyrAgene.④Of16strainscontainingmutationingyrAgene,15strainsexhibitedsubstitutionofSer91(TCC)→Phe(TTC).Conclusions①MutationswithingyrAgenemediatelowandmoderatelevelsfluoroquinoloneresistancewhilemutationswithinparCgeneparticipateinhighlevelfluoro-quinoloneresistanceinN.gonorrhoeae.②SubstitutionofSer91→PheingyrAgeneisthepivotalmutationresultinginfluoroquinoloneresistanceinN.gonorrhoeae.

Objective To investigate the blood supply of the expanded skin flap from medial up-per arm and its application in the reparation of cervical scar contracture due to sear resection. Methods The operation was carried out for three steps: (1) The expander was implanted under the superficial fascia. (2) The skin flap from medial upper arm was created with superior ulnar collateral artery as blood supply and attributive branches of basilica and axillary veins as blood collection. (3) After thes car contracture was released, the defect was covered with medial upper arm flap with maximal area of 25 cm×15 cm. Results Ten patients in all with cervical scar contracture were treated with the skin flap. All the skin flaps survived at last with nearly normal skin color, texture and contour. And the scar in donor sites seemed to be neglectable. Conclusions Reparation of cervical scar contracture with medial upper arm skin flap after expanding could be recommended. But 3 months long time and fixation of upper limb and head might be disadvantages.

Objective To construct recombinant adeno-associated virus and lentivirus carrying siRNA of TIMP-1 and to investigate the efficiency of infection and short-term inhibitory effect of TIMP-1 gene expres-sion on rat hepatic stellate cells. Methods One pair of siRNA which could effectively inhibit expression of the TIMP-1 gene in HSC-T6 was screened and cloned into AAV vector and lentiviral vector to construct the recombinant AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP. AAV/GFP and Lenti/GFP as neg-ative control were also obtained. Experiments were assigned to five groups: AAV/siRNA-TIMP-1/GFP, AAV/GFP, Lenti/siRNA-TIMP-1/GFP, Lenti/GFP group and mock group. Rat HSC-T6 cells were infected by these recombinant viruses at a concentration of MOI by 10. To monitor the efficiency of infection, fluores-cence microscope and flow cytometer were used. After 7 d post-infection, Western blot was used to detect the TIMP-1 protein expression. Results HSC-T6 had no significant changes after infection. The efficiency of infection in AAV/GFP and Lenti/GFP group were 72.7% and 70.0%, AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP group were 64.58% and 61.86%. The protein expression levels of TIMP-1 in HSC-T6 cells at 7 d post-infection by the recombinant AAV and Lentivirus were decreased 40.0% compared with those in mock control and normal HSC-T6 (P<0.05). Conclusion Recombinant AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP could effectively infect HSC-T6 with similar efficiency and suppress the expression of TIMP-1 in rat HSC-T6 remarkably.

ObjectiveTo explore the effect of sweroside on the protection of cardiac systolic/diastolic function during ischemia/reperfusion （I/R） injury. MethodTwenty-four healthy male SD rats were randomly divided into control group， model group， 10 μmol·L^-1 sweroside group and 1 μmol·L^-1 digoxin group. The I/R injury was modeled by Langendorff and ligation of the left anterior descending coronary artery. The infarct size in each group was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining and hemodynamic parameters such as left ventricular diastolic pressure （LVDP）， left ventricular end-diastolic pressure （LVEDP）， left ventricular end-systolic pressure （LVESP）， maximum rate of rising of left ventricular pressure （+dp/dt_max） and maximum rate of decreasing of left ventricular pressure （-dp/dt_max） of rat isolated heart were detected by Powerlab. In addition， neonatal rat cardiomyocytes （NRCMs） were isolated and randomly divided into control group， model group， 1 μmol·L^-1 sweroside group and 10 μmol·L^-1 sweroside group. Hypoxia/reoxygenation （H/R） injury model was established. Cardiac systolic function and calcium transients were examined by multi-functional cell imaging analyzer and laser confocal microscope. Furthermore， real-time polymerase chain reaction（Real-time PCR） was used to verify the mRNA expression of excitation-contraction coupling genes such as L-type calcium channel （Cacnb2）， cytochrome c oxidase subunit 6A2 （Cox6a2）， troponin （Tnnc1， Tnni3， Tnnt2）， actin （Actc1）， and myosin （Myh6， Myl2， Myl4） according to the results of previous transcriptome sequencing and literature investigation. Differentially expressed genes were subjected to cluster analysis. ResultCompared with the conditions in the control group， increased cardiac infarction size （P<0.01） and LVEDP （P<0.01） and decreased LVDP （P<0.01） and LVESP （P<0.05） were observed in the model group， with +dp/dt_max of increasing trend while -dp/dt_max decreasing. Moreover， the cell viability， heart rate and contraction amplitude of NRCMs was reduced （P<0.01）， while the contraction duration， time to peak and relaxation time was elevated （P<0.01） in the model group. Interestingly， sweroside could reverse these indicators （P<0.05）. In addition， the expression of Cacnb2， Cox6a2， Tnnc1， Tnni3， Tnnt2， Actc1， and Myh6， Myl2， and Myl4 was down-regulated in the model group （P<0.05， P<0.01）， but sweroside could up-regulate the expression of the above genes （P<0.05）. ConclusionSweroside effectively regulated Ca²⁺ level in NRCMs， enhanced cardiac systolic function， and protected against H/R injury by regulating excitation-contraction coupling.

ObjectiveTo establish and evaluate a mouse model of heart failure with Qi deficiency syndrome. MethodForty-four KM mice were randomly divided into sham operation group， model group， and modified Si Junzitang group （12.89 g·kg^-1）. The model group and the modified Si Junzitang group underwent thoracic aortic constriction （TAC）， while the sham operation group only underwent suture without constriction. Echocardiography and pathological examination were used to assess the heart failure model and evaluate the pharmacological effects. Macroscopic characterization， microscopic biology， and formula identification were conducted to collect general signs， body weight， open-field behavior， grip strength， mitochondrial ultrastructure， and other macroscopic and microscopic characteristics of mice. Mitochondrial fission and fusion protein expression were measured to determine the syndrome type. ResultEight weeks after TAC， compared with the sham operation group， the model group showed a significant decrease in left ventricular ejection fraction （LVEF） （P<0.01）， and modified Si Junzitang improved LVEF in mice （P<0.05）. Hematoxylin-eosin （HE） staining of the heart showed inflammatory cell infiltration and thickening of blood vessel walls in the model group， which was significantly improved by modified Si Junzitang. After 6-8 weeks， compared with the sham operation group and the modified Si Junzitang group， the model group exhibited significant hair loss， hair yellowing， decreased activity， and depression. Moreover， compared with the sham operation group， the model group had a significantly lower increase in body weight （P<0.05）， while the modified Si Junzitang group showed a significant increase in body weight （P<0.05） compared with the model group. After 6-8 weeks， compared with the sham operation group， the model group showed a significant decrease in open-field distance and speed （P<0.05）， while the modified Si Junzitang group exhibited significantly improved open-field distance and speed in the 8^th week （P<0.05）. After 6-8 weeks， compared with the sham operation group， the model group exhibited a significant decrease in maximum grip strength （P<0.05）， while the modified Si Junzitang group showed a significant increase in maximum grip strength 8 weeks after TAC （P<0.05）. Transmission electron microscopy of the gastrocnemius muscle showed uneven muscle tissue matrix， mitochondrial swelling， increased volume， matrix dissolution， ridge loss， and vacuolization in the model group， while modified Si Junzitang improved mitochondrial swelling， ridge fracture， and matrix vacuolization. Western blot analysis showed that the expression of the kinetic associated protein 1 （DRP1） in the gastrocnemius muscle of the model group significantly increased （P<0.01）， and the expression of mitochondrial fusion hormone 1 （MFN1） significantly decreased （P<0.05） as compared with those in the sham operation group. Furthermore， compared with the model group， the modified Si Junzitang group exhibited a significant decrease in the expression of DRP1 （P<0.05） and a significant increase in MFN1 expression （P<0.01）. ConclusionMice exhibited significant manifestations of qi deficiency syndrome 6-8 weeks after TAC， accompanied by abnormal mitochondrial morphology and function in the gastrocnemius muscle， which were significantly improved by modified Si Junzitang.

Objective To establish a mathematical prediction model for chronic obstructive pulmonary disease (COPD) by applying an artificial neural network (ANN) and logistic regression analysis method. Methods A cross-sectional survey was conducted in 2015 to collect epidemiological data of COPD of 2 400 residents from Hubei Province. Subjects were randomized into training group and test group at a ratio of 7:3. The prediction models of COPD were established using ANN and logistic multiple regression. The predictive performance of the two models was compared. Results Information from a total of 1 569 subjects was valid and analyzed, including 1,099 cases in the training group and 470 cases in the test group. The area under curve (AUC) of ANN for training group and test group was 0.80 and 0.78, respectively. The AUC of logistic regression for training group and test group was 0.75 and 0.74, respectively. Conclusion It is feasible to apply ANN and logistic regression models to predict COPD, which can provide scientific evidence for COPD prevention and treatment.