Objective To clarify the effects of Xinfukang containing-serum on stromal cell-derived factor-1α (SDF-1α) translation and protein secretion of bone marrow stem cells (BMSCs).Methods BMSCs were isolated and amplified using bone marrow culture method,and were identified by flow cytometry.mRNA and protein secretion of SDF-1α were detected by quantitative PCR (q-PCR) and enzyme linked immunosorbent assay (ELISA),respectively.Results The expression of SDF-1α mRNA were significantly increased after 72 h in drug-containing serum,and SDF-1α mRNA in the experimental group was approximately 200 times as that in the control group (P＜0.05).Secretion of SDF-1 α in the experimental group (277.561 1 ± 15.651 8) pg/ml was nearly doubled compared with that in the control group (153.107 1±14.765 1) pg/ml (P＜0.05).Conclusions BMSCs from whole bone marrow adherent culture have high purity,and drug-containing serum can promote BMSCs to express SDF-1 α mRNA and secretion of SDF-1 α.

Two pedigrees are reported here including two siblings and a boy who were diagnosed with Aicardi-Goutières syndrome type 3 (AGS3) caused by compound heterozygous variation of RNASEH2C gene. Prenatal gene diagnosis was performed when their mothers were pregnant again. All three cases presented with epilepsy, microcephaly, muscular hypertonia and severe language, motor and mental retardation. In pedigree 1, genetic analysis showed compound heterozygous variants of c.194G>A (p.Gly65Asp) and c.434G>T (p.Arg145Leu) in the RNASEH2C gene of proband 1 and her younger brother, which were inherited from their mother and father respectively. While in pedigree 2, c.194G>A(p.Gly65Asp) and c.227C>T(p.Pro76Leu) compound heterozygous variants in the RNASEH2C gene were found in proband 2, which were inherited from his father and mother, respectively. Diagnosis of AGS3 was confirmed in these three cases based on their medical history and the testing results. The mothers from the two families underwent prenatal diagnosis in their subsequent pregnancy, and the variation only inherited from the mothers was detected, suggesting that the two fetuses are carriers. Both families chose to continue the pregnancy and delivered at full-term. No growth or development abnormalities were reported in the children during a one-year follow-up.

Objective To assess the clinical significance of extra-adipose capsule route and intra-adipose capsule route for the resection of benign adrenal tumors with retroperitoneal laparoscopic adrenalectomy(RLA),and to explore the selection of route based on the mayo adhesive probability(MAP)scoring system.Methods Clinical data of 102 patients who received RLA and pathologically diagnosed as benign adrenal tumors during Feb.2015 and Dec.2020 in the First Affiliated Hospital of Xi'an Jiaotong University were retrospectively reviewed,and MAP scores were assessed with the preoperative tomography images.The gross and MAP score stratified perioperative outcomes between extra-adipose capsule route(classical group,n=56)and intra-adipose capsule route(modified group,n=46)were compared respectively.Results All procedures were successfully completed with no conversion to open surgery and with no need for transfusion.There were no significant differences in operation time[(102.1±26.3)min vs.(110.2±32.1)min,P=0.17]and intraoperative blood loss[(53.5±34.0)mL vs.(61.1±48.4)mL,P=0.35]between the two groups.Subgroup analysis based on MAP score showed that for low risk patients(MAP score 0-2),operative results were comparable between the two groups,but for high risk patients(MAP score 3-5),the operation time was significantly shorter[(114.7±20.7)min vs.(137.2±23.0)min,P＜0.01],and blood loss was significantly less[(52.7±33.1)mL vs.(92.8±49.7)mL,P=0.01]in the classical group than in the modified group.Conclusion RLA could be performed with either surgical routes safely and effectively.MAP scoring system could be an effective tool for preoperative surgical route planning.Compared with the modified route,classical route is more suitable for patients with high MAP score to achieve better operative outcomes.

Objective : To explore whether miR⁃9 affects the proliferation of neural stem cells (NSCs) by regulating β ⁃tubulin Ⅲ and glial fibrillary acidic protein (GFAP) . Methods : NSCs cells were isolated and cultured on the purchased healthy pregnant mice and the isolated cells were divided into NO group ( non⁃transfected NSCs cell line) , NN group (NSCs transfected with miR⁃9 ⁃NC) , NM group (NSCs transfected) transfected with miR⁃9 mimics) , NI group ( transfected with miR⁃9 inhibitor) . Nestin was identified by immunofluorescence , the contents of Mir⁃9 , β ⁃tubulin Ⅲ and GFAP were measured by QRT⁃PCR , the proliferation of NSCs was measured by MTT method , the apoptosis of NSCs was measured by flow cytometry , and the protein expressions of Mir⁃9 , β ⁃tubulin Ⅲ and GFAP were detected by Western blot. Results : The detection index Nestin of NSCs for 24 h after cell passage was positive in 90% of the NSCs , and co⁃localized with the nucleus , indicating the successful isolation of NSCs. The expression of miR⁃9 mRNA in NSCs of the NM group was the highest among the four groups , indicating that the transfection was successful. Among the four groups , β ⁃tubulin Ⅲ mRNA and protein expression was the highest in the NM group , and the GFAP mRNA and protein expression was the lowest. The NI group was the opposite (P < 0. 05) . In the four groups after 24 h , the cell activity of NSCs in the NM group was the strongest and showed a trend of gradually increasing with time (P < 0. 05) . The cell activity of NI was significantly weaker than that of the other three groups (P < 0. 05) . Among the four groups , the apoptosis rate of the NM group was lower than that of the other three groups (P < 0. 05) , and the apoptosis rate of NI was higher than that of the other three groups (P < 0. 05) . The difference in the above indicators between the NO group and the NN group was smaller. Conclusion Overexpressed miR⁃9 promotes the proliferation of NSCs by promoting the expression of β ⁃tubulin Ⅲ and inhibiting the expression of GFAP.

ObjectiveTo investigate the mechanism of Naoxintong capsules on ischemia-reperfusion （I/R） injury in rats based on the changes of pericytes mediated by Ras homolog family member A （RHOA）/Rho-associated coiled-coil containing protein kinase 1 （ROCK1） pathway. MethodsNinety rats （15 rats for each group） were randomly divided into a sham operation group， a model group， a positive control group receiving Ginkgo biloba extract （21.6 mg·kg^-1）， and groups receiving Naoxintong capsules at low， medium， and high doses of 55， 110， and 220 mg·kg^-1 （NXT-L， NXT-M， and NXT-H groups）， respectively. Except for those in the sham operation group， all rats were subjected to transient middle cerebral artery occlusion （tMCAO） to establish the experiment model. Nerve function was assessed using a neurological function score. Cerebral blood flow was detected using a laser speckle contrast imager， and the cerebral infarction rate was calculated using 2，3，5-Triphenyl tetrazolium chloride （TTC） staining. Pathological changes were observed by hematoxylin-eosin （HE） staining and Nissl staining， while pericyte morphology was observed via transmission electron microscopy. Blood-brain barrier destruction was observed by Evans blue staining. Albumin and ischemia-modified albumin levels were measured using assay kits. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the mRNA and protein expression levels of RHOA， ROCK1， platelet-derived growth factor receptor β （PDGFRB）， α-smooth muscle actin （α-SMA）， tight junction protein （ZO-1）， matrix metalloproteinase-2 （MMP-2）， and matrix metalloproteinase-9 （MMP-9）. ResultsCompared with the sham operation group， the model group exhibited decreased neurological function scores， higher percentage reduction in blood flow， and increased cerebral infarction rates （P<0.01）. Additionally， cortical neuronal nucleus shrinkage， edema， a decreased number of Nissl bodies， reduced pericyte area， elevated albumin content in the cortex （P<0.05）， and increased ischemic modified albumin levels （P<0.01） were observed. The mRNA and protein expression levels of RHOA， ROCK1， PDGFRB， α-SMA， MMP-2， and MMP-9 were increased （P<0.01）， while those of ZO-1 were decreased. Compared with the model group， all treatment groups showed improved neurological function scores， lower percentage reduction in blood flow， reduced cerebral infarction rates （P<0.01）， alleviated cortical histological changes， increased number of Nissl bodies， expanded pericyte area， decreased albumin content in the cortex， and reduced ischemia-modified albumin levels （P<0.01）. The mRNA and protein expression levels of RHOA， ROCK1， PDGFRB， α-SMA， MMP-2， and MMP-9 were decreased （P<0.01）， while those of ZO-1 were increased. Among the treatment groups， the NXT-M group showed the most pronounced improvement in cerebral I/R injury. ConclusionNaoxintong capsules can restore cerebral blood supply， reduce microcirculation disturbance， and protect blood-brain barrier in rats with I/R injury. Its mechanism of action may be related to the inhibition of the RHOA/ROCK1 signaling pathway and reduced pericyte contraction.

ObjectiveTo investigate the mechanism of Naoxintong capsules on ischemia-reperfusion （I/R） injury in rats based on the changes of pericytes mediated by Ras homolog family member A （RHOA）/Rho-associated coiled-coil containing protein kinase 1 （ROCK1） pathway. MethodsNinety rats （15 rats for each group） were randomly divided into a sham operation group， a model group， a positive control group receiving Ginkgo biloba extract （21.6 mg·kg^-1）， and groups receiving Naoxintong capsules at low， medium， and high doses of 55， 110， and 220 mg·kg^-1 （NXT-L， NXT-M， and NXT-H groups）， respectively. Except for those in the sham operation group， all rats were subjected to transient middle cerebral artery occlusion （tMCAO） to establish the experiment model. Nerve function was assessed using a neurological function score. Cerebral blood flow was detected using a laser speckle contrast imager， and the cerebral infarction rate was calculated using 2，3，5-Triphenyl tetrazolium chloride （TTC） staining. Pathological changes were observed by hematoxylin-eosin （HE） staining and Nissl staining， while pericyte morphology was observed via transmission electron microscopy. Blood-brain barrier destruction was observed by Evans blue staining. Albumin and ischemia-modified albumin levels were measured using assay kits. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the mRNA and protein expression levels of RHOA， ROCK1， platelet-derived growth factor receptor β （PDGFRB）， α-smooth muscle actin （α-SMA）， tight junction protein （ZO-1）， matrix metalloproteinase-2 （MMP-2）， and matrix metalloproteinase-9 （MMP-9）. ResultsCompared with the sham operation group， the model group exhibited decreased neurological function scores， higher percentage reduction in blood flow， and increased cerebral infarction rates （P<0.01）. Additionally， cortical neuronal nucleus shrinkage， edema， a decreased number of Nissl bodies， reduced pericyte area， elevated albumin content in the cortex （P<0.05）， and increased ischemic modified albumin levels （P<0.01） were observed. The mRNA and protein expression levels of RHOA， ROCK1， PDGFRB， α-SMA， MMP-2， and MMP-9 were increased （P<0.01）， while those of ZO-1 were decreased. Compared with the model group， all treatment groups showed improved neurological function scores， lower percentage reduction in blood flow， reduced cerebral infarction rates （P<0.01）， alleviated cortical histological changes， increased number of Nissl bodies， expanded pericyte area， decreased albumin content in the cortex， and reduced ischemia-modified albumin levels （P<0.01）. The mRNA and protein expression levels of RHOA， ROCK1， PDGFRB， α-SMA， MMP-2， and MMP-9 were decreased （P<0.01）， while those of ZO-1 were increased. Among the treatment groups， the NXT-M group showed the most pronounced improvement in cerebral I/R injury. ConclusionNaoxintong capsules can restore cerebral blood supply， reduce microcirculation disturbance， and protect blood-brain barrier in rats with I/R injury. Its mechanism of action may be related to the inhibition of the RHOA/ROCK1 signaling pathway and reduced pericyte contraction.