BACKGROUND:Endothelial injury is one of the causes of cardiovascular diseases.Human induced pluripotent stem cells are easy to obtain,have strong differentiation ability,and have less exclusiveness,and their endothelial differentiated cells can be used as ideal cells for cardiovascular disease research. OBJECTIVE:To investigate the effect and mechanism of calycosin on endothelial differentiation of human induced pluripotent stem cells and to provide technical support for microvascular regeneration. METHODS:Human induced pluripotent stem cells were divided into control group and calycosin group(1.25,2.5 μg/mL),and growth factors were added to induce single-layer endothelial differentiation.After the induction of differentiation for 8 days,the positive rate of endothelial cell marker CD144 was detected by flow cytometry.Fluorescent expressions of CD144 and CD31 were detected by the immunofluorescence method.Lentivirus RNAi GFP puromycin was used to silence human-induced pluripotent stem cell Piezo1 mRNA followed by endothelial directed differentiation.After 8 days of differentiation,the positive rate of CD144 in differentiated cells was detected by flow cytometry.The mRNA expression levels of CD144,Piezo1 and MEK were detected by qPCR. RESULTS AND CONCLUSION:(1)Compared with the control group,the positive rate of CD144 was significantly increased in the 1.25 and 2.5 μg/mL calycosin groups(P<0.05).The expressions of CD144,Piezo1,and MEK mRNA were increased in the 2.5 μg/mL calycosin group(P<0.05).The fluorescence expressions of CD144(P<0.01)and CD31(P<0.001)were significantly increased in the 2.5 μg/mL calycosin group.(2)Compared with the shNT group,CD144 positive rate and CD144,Piezo1,MEK mRNA expressions were significantly increased in the shNT + calycosin 1.25,2.5 μg/mL groups(P<0.05).Compared with the shPiezo1 group,the positive rate of CD144 and mRNA expressions of CD144,Piezo1 and MEK had no significant changes in the shPiezo1+calycosin 1.25,2.5 μg/mL groups(P>0.05).(3)It is concluded that 2.5 μg/mL calycosin promotes the differentiation of human-induced pluripotent stem cells into endothelial lineages.Calycosin promotes the downstream MEK expression,thereby promoting the endothelial differentiation of human induced pluripotent stem cells by targeting the expression level of Piezo1.

The most prevalent kind of renal calculi,calcium oxalate(CaOx),is characterized by its propensi-ty for recurrence in the urinary system.The development of CaOx renal calculi is greatly affected by macrophage polariza-tion.Particular oxalate causes an imbalance in macrophage polarization,which skews the M1/M2 ratio and makes it easier for CaOx crystals to accumulate in the kidneys and grow into calcium plaques in the renal papilla.Notably,M2 macro-phages can prevent CaOx renal calculi by consuming crystals and reducing inflammatory stress.As a result,immunothera-peutic techniques that alter M1 and M2 macrophage polarization are extremely promising for preventing CaOx renal calcu-li.To clarify the respective roles of M1 and M2 macrophages in the formation of CaOx crystals and provide insights for de-veloping immunotherapeutic interventions against CaOx renal calculi,this review summarizes the mechanisms underlying macrophage polarization in the genesis of CaOx renal calculi.

Objective To evaluate the accuracy of confocal laser endomicroscopy (CLE)in primary bile reflux gastritis (BRG).Methods From November 10th to December 15th,2015 ,55 patients underwent CLE examination and preliminarily diagnosed as BRG with traditional white-light endoscopy were enrolled.CLE score standard was designed.Dixon pathologic score was considered as gold standard. Receiver operating characteristic (ROC)curve was drawn to evaluate the accuracy of CLE score in BRG diagnosis.Sensitivity,specificity and 95 % confidence interval (CI )were calculated.Kappa analysis was performed to assess the inter-observer agreement of CLE score.Results According to Dixon pathologic score standard,29 patients (52.7%)were diagnosed as primary BRG among the 55 enrolled patients. Among the 42 Helicobacter pylori (H .pylori )negative patients,the area under receiver operating characteristic curve (AUC)of CLE in BRG diagnosis was 0.90 (95 %CI 0.81 —1 .00).Taking CLE score over six as the cut-off value for diagnosis,the sensitivity and specificity was 84.00% (95 %CI 65 .35 %—93.60%)and 82.35 % (95 %CI 58.97%—93.81 %),respectively.The Kappa value for inter-observer agreement in BRG diagnosis was 0.60 (95 %CI 0.24—0.95).Conclusion Primary BRG can be accurately diagnosed by CLE in H .pylori negative patients with high sensitivity and specificity.

Objective To analyze the pattern of normal bone marrow on whole body diffusionweighted imaging (WB-DWI) and its influence factors.MethodsA total of 98 healthy volunteers (male 47 and female 51). All volunteers underwent WB-DWI at 1.5 T MR scan. The ADC value,the signal intensity on DWI obtained with a b value of 800 s/mm2 ( SIDWI ),and the signal intensity on short time inversion recovery images ( SISTIR ) of thoracic vertebrae,lumber vertebrae,bilateral head of femur,bilateral neck of femur,bilateral superior segment of femur,bilateral inferior segment of femur,bilateral ilium,bilateral head of humerus,bilateral scapula were measured and compared with ANOVA test and StudentNewmar-Keuls test.The normal appearance of bone marrow on WB-DW[ was assessed. The relationship between the SIDwI and the ADC,and the SIDWI and the SISTIR of bone marrow were analyzed.The ADC of bone marrow between male and femaIe were compared. Spearman correlation analysis was performed for different age groups.Results( 1 ) Bone marrow signal intensity was different among 98 healthy volunteers.Bone marrow in 69 healthy volunteers (female 24,male 45 ) showed low to intermediate signal intensity,whereas in the remaining 29 healthy volunteers (female 27,male 2) showed high signal intensity.(2) The SIDWI of thoracic vertebrae ( median 44.54),lumber vertebrae ( median 35.01 ),head of femur ( median 13.61 ),neck of femur ( median 16.00),superior segment of femur ( median 21.45 ),ilium ( median 25.77),head of humerus (median 18.35),scapula (median 36.12) was positively correlated with the ADC [ (0.55 ±0.08) × l0-3,(0.53 ±0.08) × 10-3,(0.30 ± 0.10) × 10-3,(0.42 ± 0.16) × 10-3,(0.74±0.14) ×l0- 3,(0.49±0.10) ×10-3,(0.36±0.13) ×10-3,(0.49±0.11) × 10-3mm2/s]and the SISTIR ( median 61.81,64.99,53.27,69.08,73.10,66.35,73.16,79.81 ),r =0.513 and 0.695,0.741 and 0.764,0.443 and 0.489,0.641 and 0.656,0.510 and 0.648,0.475 and 0.715,0.366 and 0.446,0.437 and 0.739 ;P ＜ 0.01. (3) There was significant difference of the ADC of bone marrow in different bone,F =138.69,P ＜ 0.01. Student-Newman-Keuls test revealed that no significant difference was found in the ADC between thoracic vertebrae and lumbar vertebrae,ilium and scapula,head of humerus and inferior segment of femur ( P ＞ 0.05 ),and significant difference was found in the ADC values between the remaining two groups ( P ＜ 0.05 ). The bones associated with decreasing ADC values were superior segment of femur,thoracic vertebrae and lumber vertebrae,ilium and scapula,neck of femur,head of humerus,head of femur and inferior segment of femur. ( 4 ) The ADC values of bone marrow of female subjects in thoracic vertebrae [ (0.59 ±0.07) × 10-3 mm2/s],lumber vertebrae [ (0.58 ±0.06) × 10 -3 mm2/s],head of femur ( median 0.33 × 10 -3 mm2/s),neck of femur ( median 0.53 × 10 -3 mm2/s),superior segment of femur ( median 0.81 × 10-3 mm2/s),inferior segment of femur ( median 0.32 ×10-3 mm2/s),ilium [ (0.52 ± 0.09 ) × 10-3 mm2/s ],head of humerus (median 0.42 × 10-3 mm2/s),scapula [ (0.53 ± 0.09) × 10-3 mm2/s] were significantly higher than those of male subjects [ (0.51 ±0.07) × 10-3,(0.48 ±0.07) × 10-3,median 0.23 × 10-3,median 0.31 × 10-3,median 0.66 × 10-3,median 0.23 × 10-3,(0.46 ±0.10) × 10-3,median 0.27 × 10-3,(0.45 ±0.11 ) × 10 3mm2/s].(5)There was significant negative correlation between the ADC values of bone marrow and age in thoracic vertebrae,lumber vertebrae,head of femur,neck of femur,superior segment of femur,ilium,head of humerus for female subjects,r =-0.549, -0.629, -0.329, -0.524, -0.338, -0.548 and -0.416,respectively,P ＜ 0.05.There was no significant correlation between ADC values and age in inferior segment of femur and scapula for female subjects and all the regions for male subjects ( P ＞ 0.05 ).Conclusions The ADC and the SIsTIR of bone marrow correlates with the SIDW1.The ADC values of bone marrow is affected by age and sex,and is different for different bones.

Objective To evaluate the effect and safety of thymosin combined with biapenem in the treatment of severe lower respiratory tract infection of gram negative bacilli.Methods 54 patients with severe lower respiratory tract infection of gram negative bacilli were randomly divided into study group (group A) and control group (group B),group A was treated with thymosin and biapenem,group B biapenem.All patients were treated for 7 ～ 14 days in a course.The clinical effect,treatment time of biapenem,the bacterial clearance rate and adverse drug reaction betweeen two groups were compared.Results The clinical effective rate of group A and B were 84.6％ and 82.1％,respectively,there was no statistical difference (P ＞ 0.05).Treatment time of biapenem in group A and B was (8.2 ±3.4) days and (13.2 ± 3.6) days,respectively,there was statistical difference (P ＜ 0.05).The bacterial clearance rate of group A and B were 86.7％ and 80.7％,respectively,there was no statistical difference between two groups (P ＞ 0.05).Conclusion The treatment time of thymosin combined with biapenem treatment of gram-negative bacilli severe respiratory tract infection was shorter than that of biapenem alone treatment,and it was safe,effective,and worthy of ciinical application.

A new artificial somatic-autonomic neuroanastomosis has been established in male rats with spinal cord injury (SCI). Anorectal manometry and neural retrograde tracing were conducted in this animal model to analyze the mechanisms and the effects on recovery of anorectal function. The left L4 ventral root (L4VR) was intradurally micro-anastomosed to the L6 ventral root (L6VR) to establish the new regenerated neural pathway. Three months later the spinal cord was completely transected at the T9-10 level. Eight weeks later the model rats were randomly divided into two groups. The rats in the group 1 (n=8) were applied for anorectal manometry, and those in the group 2 (n=4) were used for neural retrograde tracing study with fluorogold (FG) and dextran tetramethylrhodamine (TMR). The results of anorectal manometry showed the new reflex pathway could induce rectum to contract and simultaneously electric activity of external anal sphincter (EAS) to become weak or disappearing (indicating synergetic relaxation of EAS). FG and TMR dual labeled neurons with round and elliptical shape were mainly observed in L4 angulus anterior of model rats. The regenerated neural pathways were effective to improve the rectum external sphincter synergetic status and restore the anorectal function.

A new artificial somatic-autonomic neuroanastomosis has been established in male rats with spinal cord injury(SCI).Anorectal manometry and neural retrograde tracing were conducted in this animal model to analyze the mechanisms and the effects on recovery of anorectal function.The left L4 ventral root(L4VR)was intradurally micro-anastomosed to the L6 ventral root(L6VR)to establishthe new regenerated neural pathway.Three months later the spinal cord was completely transected at the T9-10 level.Eight weeks later the model rats were randomly divided into two groups.The rats in the group 1(n=8)were applied for anorectal manometry,and those in the group 2(n=4)were used for neural retrograde tracing study with fluorogold(FG)and dextran tetramethylrhodamine(TMR).The results of anorectal manometry showed the new reflex pathway could induce rectum to contract and simultaneously electric activity of external anal sphincter(EAS)to become weak or disappearing (indicating synergetic relaxation of EAS).FG and TMR dual labeled neurons with round and elliptical shape were mainly observed in L4 angulus anterior of model rats.The regenerated neural pathways were effective to improve the rectum external sphincter synergetie status and restore the anorectal function.

A protocol for the isolation, purification and culture of motor neurons from newborn rat spinal cord was described and the effect of glial cell line-derived neurotrophic factor (GDNF) on the growth of neurite of motor neurons was investigated in vitro. Spinal motor neurons (SMNs) were dissociated from ventral spinal cord of postnatal day 1 rats. The culture system for SMNs was established by density gradient centrifugation, differential adhesion, and use of serum-free defined media and addition of exogenous GDNF. After 72-h culture, the cells displayed the characteristic morphology of motor neurons, exhibited extensive neuritic processes and were positive for choline acetyltransferase (ChAT) expression. The neurite length of SMNs in GDNF groups was significantly longer than that in control group (P<0.05). This protocol can be adapted for various postnatal motor neurons studies.

ificantly longer than that in control group (P<0.05). This protocol can be adapted for various postnatal motor neurons studies.

Objective:To study the mechanism of stem brain functional reorganization in a spinal bifida patient compli-cated with congenital urinary incontinence after treatment with artificial nerve arc. Methods=Compared the stem brain func-tional map in the patient pre-and post-operatively with functional magnetic resonance imaging (fMRI). Results: No activa-tion was found in the patient before operation while strong activation was found after operation. Conclusions: The micturi-tion center was formed in the stem brain after construction of artificial nerve arc.