BACKGROUND:Astrocytes are the most abundant cells in the central nervous system,and various subsets of astrocytes are heterogeneous,performing a variety of special functions.Single-cell RNA sequencing(scRNA-seq)technology developed in recent years has extended our understanding of astrocyte heterogeneity from the perspective of transcriptome profiling. OBJECTIVE:To summarize the heterogeneity of scRNA-seq technology in different time and space,and pathological states and expand our knowledge of astrocyte heterogeneity on both molecular and functional levels. METHODS:The relevant articles on astrocyte heterogeneity and scRNA-seq were searched on PubMed,Elsevier,and CNKI databases.The search terms were"astrocytes,scRNA-seq,heterogeneity,Alzheimer disease,spinal cord injury,multiple sclerosis"in Chinese and English.Finally,74 articles were selected for viewing after screening according to inclusion criteria. RESULTS AND CONCLUSION:scRNA-seq studies related to the heterogeneity of astrocytes have shown that astrocyte is significantly heterogeneous across four aspects:species,developmental stage,central nervous system region,and pathological state.(1)Unique expression of certain genes occurs in astrocytes of different species,and the discovery of species-specific genes is beneficial for the translation of clinical studies.(2)During astrocyte development,differential gene expression emerged in the cellular subtypes identified at each stage,which further refined the cellular lineage of astrocytes and laid the foundation for the study of astrocyte developmental trajectories and mechanisms.(3)The discovery of differential gene expression allows regional localization of different astrocyte subpopulations and assists in the diagnosis and treatment of neurological diseases.(4)Astrocyte heterogeneity revealed by scRNA-seq can provide specific markers at the time of disease diagnosis and identify potential therapeutic targets.(5)The heterogeneity of astrocytes exists in many aspects,interacts with each other and is complex.The mechanisms of its generation,maintenance and transformation remain unclear.At present,molecular research on the single-cell level is still lacking.Linking transcriptionally defined astrocyte subpopulations to cellular activity,behavior and disease markers in real time remains one of the great challenges in the field.

BACKGROUND:Vascularization is essential for wound healing and functional recovery during soft tissue repair.Adipose tissue is believed to be the body's largest source of stem cells,and a number of different fat complexes have been developed for research and treatment.Its ability to promote angiogenesis and soft tissue repair has been extensively studied. OBJECTIVE:To review the progress of vascularization in soft tissue repair,and to summarize the preparation methods of adipose tissue and its derivative and their applications in vascularization and soft tissue repair.It is proven that adipose tissue and its derivative have excellent research value and clinical application prospects in vascular and soft tissue engineering. METHODS:PubMed,Web of Science and CNKI databases were used to search the related articles published from January 2010 to February 2023.The search terms were"soft tissue repair,wound healing,vascularization,angiogenesis,adipose tissue,stromal vascular fraction,adipose tissue-derived microvascular fragment,nanofat,adipose extracellular matrix/stromal vascular fraction gel"in Chinese and English.A small number of old classic literature was also included.An initial screening was performed by reading the titles and abstracts to exclude literature that was not relevant to the topic of the article,and 69 papers were finally included for the analysis of the result. RESULTS AND CONCLUSION:(1)Wound healing is an important physiological process,which mainly occurs when tissue is damaged,such as injury,surgery,burn,tumor,infection and vascular disease caused by tissue damage and defects.(2)Adequate vascularization of the wound site is essential for tissue repair,reconstruction of local homeostasis and functional recovery.(3)Adipose tissue is believed to be the body's largest pool of stem cells and a number of different fat components have been used for research and treatment.(4)Due to its inherent composition and preparation advantages,adipose tissue will continue to play an important role in tissue engineering research and therapy.

Objective:To investigate the risk factors of positive peritoneal cytology (PPC) in patients with endometrial cancer and the impact of PPC on patients' prognosis.Methods:The clinicopathological data of 202 patients who underwent initial surgical treatment and were diagnosed with endometrial cancer by postoperative pathology at Qilu Hospital of Shandong University from January 2015 to December 2019 were retrospectively analyzed, and the peritoneal fluid of patients were sent intraoperatively for cytological liquid-based smear examination. Logistic regression was used to perform univariate and multivariate analyses of PPC in the whole group of patients and the early-stage patients; Univariate analysis of the progression-free survival in the whole group of patients and the early-stage patients was performed by Kaplan-Meier method and compared by log-rank method, and multivariate analysis of the progression-free survival in the whole group of patients and the early-stage patients was performed by Cox proportional hazards model.Results:Of 202 patients, 183 (90.6%) had negative peritoneal cytology (NPC) and 19 (9.4%) had PPC; 180 patients (89.1%) were stage Ⅰ-Ⅱ and 22 (10.9%) were stage Ⅲ-Ⅳ; 180 patients (89.1%) had early-stage endometrial cancer. Deep myometrial infiltration ( OR = 3.57, 95% CI 1.02-12.45, P = 0.046) and lymph node metastasis ( OR = 7.16, 95% CI 1.70-30.23, P = 0.007) were independent risk factors for PPC in patients with endometrial cancer; deep myometrial infiltration was an independent risk factor for PPC in patients with early-stage endometrial cancer ( OR = 6.22, 95% CI 1.22-31.73, P = 0.028). The 3-year PFS rates for the whole group of patients with PPC and NPC were 72.9% and 92.7%, and the difference was statistically significant ( P = 0.001); the 3-year PFS rates for early-stage patients with PPC and NPC were 82.5% and 96.2%, and the difference was statistically significant ( P = 0.002). PPC was an independent risk factor for PFS in the whole group of patients with endometrial cancer ( HR = 4.80, 95% CI 1.14-20.17, P=0.032); PPC was also an independent risk factor for PFS in patients with early-stage endometrial cancer ( HR = 8.85, 95% CI 1.96-39.93, P = 0.005). Conclusions:Deep myometrial infiltration is an independent risk factor for PPC, and PPC is an independent risk factor for PFS in patients with endometrial cancer. Routine cytological examination of peritoneal fluid is recommended in patients with endometrial cancer.

Objective To establish an in vitro cell model of Parkinson disease with SHSY5Y cells over-expressing human A53T mutant alpha-synuclein and to examine the effects of Aβ1-42 oligomer on cell survival and autophagy function in the cell model Method The recombinant lentivirus containing the A53T mutant alpha-synuclein gene or empty vector were transfected to SHSY5Y cells. The expression of α-synuclein mRNA in SHSY5Y cells was detected by RT-qPCR. The effect of Aβ1-42 oligomer on cell proliferation was detected with CCK-8 after incubation with Aβ1-42 oligomer for 24 hours. The autophagy-related proteins were evaluated with Western Blot. Result The mRNA and protein levels of alpha-synuclein were significantly increased in SHSY5Y cells expressing alpha-synuclein. There were no significant difference in the cell proliferation between alpha-synuclein group and control group (P<0.001) . Incubation with Aβ1-42 oligomer significantly decreased the proliferation rate in alpha-synuclein group in a dose-dependent manner compared with the control group. The levels of autophagy related proteins including LC3-Ⅱ and Beclin-1 were significantly lower in alpha-synuclein group than in control group (P<0.05). Conclusion This work has constructed an in vitro cell model of Parkinson′s disease. The over-expression of A53T mutant alpha-synuclein do not affect the cell survival whereas the Aβ1-42 oligomer exhibits toxic effects on cells expressing alpha-synuclein possible through suppression of the autophagy activation.

Objective To analyze the correlation between the direct measurement of pulmonary artery pressure and the related echocardiographic parameters in rats with pulmonary arterial hypertension ( PA H ) , and establish a predictable equation for pulmonary artery pressure using non‐invasive ultrasonic parameters . Methods Fifteen male Wistar rats were randomly divided into normal control ( NC ) group with five rats and PA H model group with 10 rats .PA H model was established by intraperitoneal injection of 1% MCT solution in the dose of 60 mg/kg . All the rats were examined by ultrasonic apparatus to record cardiac parameters including right ventricle anterior wall thickness ( RVAWT ) ,pulmonary artery diameter ( PAD) , aorta diameter ( AOD ) , pulmonary artery acceleration time ( PAAT ) , pulmonary artery ejection time ( PAET ) ,right ventricle end‐diastolic diameter ( RVEDD ) ,right ventricle end‐diastolic length ( RVEDL ) , tricuspid annular plane systolic excursion ( T APSE) and left ventricular ejection fraction ( LVEF ) before experiments as well as 2 and 4 weeks after modeling . At the fifth week of modeling ,all the rats were administrated with thoracotomy and right ventricular catheter to obtain pulmonary artery systolic ,diastolic and mean pressures ( PASP ,PADP and PAM P) . Results As time went on ,measures of RVAWT ,PAD , PAD/AOD ,RVEDD ,RVEDL ,RVEDD/RVEDL increased ,while measurements of PAA T ,PAA T/PAET , T APSE decreased in the model group .T he changes of RVAWT ,PAD ,PAA T/PAET ,RVEDD in the model group appeared early in the second week in contrast to data before molding ( P ＜0 .05) . When comparing model group with NC group ,there were statistic differences of RVAWT ,PAAT/PAET as early as 2 weeks after modeling measuring (all P ＜0 .05) and the dramatic variance in the parameters of PAD/AOD ,PAAT , RVEDD ,RVEDD/RVEDL ,T APSE appeared in 4‐week observation . Correlation analysis suggested there were high‐degree correlations between PAA T ,PAA T/PAET and PASP ,PAM P ( for PASP : r ＝ －0 .829 ,－0 .865 ,P＜ 0 .05 ; for PAM P : r ＝ －0 .831 , －0 .842 , P ＜ 0 .05 ) ,and moderate‐degree correlations between RVAWT ,PAD/AOD ,RVEDD ,RVEDD/RVEDL ,T APSE and PASP ,PAM P ( for PASP :|r|＝0 .615－0 .786 , P ＜0 .05 ; for PAM P : r ＝0 .683－0 .799 , P ＜0 .05) .T he linear dependent equations were established as PASP ＝ －169 .392 PAAT/PAET + 105 .092 ( r2 ＝ 0 .748 , P ＝ 0 .000 ) ,PASP ＝ 49 .576 RVAWT+67 .314RVEDD/RVEDL －45 .198 ( r2 ＝0 .731 , P ＝0 .003) ,PAM P＝ －150 .664PAAT/PAET+88 .156 ( r2 ＝0 .709 , P ＝ 0 .001 ) ,PAM P＝37 .988RVAWT +82 .072RVEDD/RVEDL －50 .517 ( r2 ＝0 .794 , P ＝ 0 .001 ) to represent the relationships between PASP or PAM P and PAAT/PAET or RVAWTcombined RVEDD/RVEDL . Conclusions Echocardiography can monitor changes in heart structure and hemodynamics .Ultrasonic parameters especially PAAT/PAET or RVAWT ,RVEDD/RVEDL could be used to estimate PASP or PAM P measured by catheterization .

Objective To study the effects of serum C-type natriuretic peptide (CNP) ,insulin-like growth factor II (IGF-Ⅱ ) , endothelin (ET) ,neuron-specific enolase (NSE) and S100B protein(S100B) on the prognosis of the patients with traumatic brain injury .Methods A total of 110 patients with craniocerebral trauma admitted in our hospital from January 2016 to January 2017 se-lected as the craniocerebral trauma group and further divided into the mild ,moderate and severe craniocerebral trauma groups ac-cording to the Glasgow Coma Scale (GCS) .Then the levels of serum CNP ,IGF-Ⅱ ,ET ,NSE and S100B in all cases were analyzed by enzyme-linked immunosorbent assay (ELISA) .Their influence on the prognosis of the patients with craniocerebral trauma and the correlation among various indicators were analyzed .Results The levels of CNP and IGF-Ⅱat admission in the craniocerebral trauma group were significantly decreased ,while the levels of ET ,NSE and S100B were significantly increased ,the difference com-pared with the control group was statistically significant (P<0 .05) .Serum CNP and IGF-Ⅱlevels in the death group ,plant survival group and disabled group were significantly decreased .The difference was statistically significant (P<0 .01) .Serum CNP and IGF-Ⅱlevels in the moderate and severe craniocerebral trauma groups were gradually increased with the disease course progress ,while serum ET ,NSE and S100B levels were gradually decreased with the disease course progress ,the difference was statistically signifi-cant(P<0 .05) .In the patients with craniocerebral trauma ,the positive correlation existed between CNP and IGF-Ⅱ ,between ET and S100B ,between ET and NSE ,and between NSE and S100B(P<0 .01) ,while the negative correlation existed between IGF-Ⅱand ET ,between IGF-Ⅱ and S100B ,between CNP and ET ,and between IGF-Ⅱand NSE (P<0 .01) .Conclusion Serum CNP , IGF-Ⅱ ,ET ,NSE and S100B are correlated to the severity of craniocerebral trauma ,which has a higher clinical application value for judging the disease condition ,evaluating the prognosis in cradiocerebral trauma .

Objective To analyze MRI in measurement of osteosarcoma invasion medullary cavity function and determine the reasonable range of the accuracy of bone cutting plane .Methods Selected 30 osteosarcoma patients with limb-salvage surgery ,X-ray and M RI in preoperative ,postoperative naked eye tumor specimens scope of intramedullary tumor ,intraoperative bone marrow derived ,postoperative histopathological examination with tumor specimens counterparts to determine the micro range .Postoperative evaluated limb function of patients .Results The average rate of functional recovery in patients was 81% ,the range measured by X-ray was significantly less than the pathologic examination(P<0 .05) ,while MRI measured attack range similar to pathological ex-amination ,there was no significant difference .Conclusion The accuracy of scope of medullary cavity osteosarcoma invasion diag-nosed by MRI is high ,as the operation reference high reliability ,short-term follow-up showed that in intramedullary outside the boundary of a 30 mm as bone cutting plane is safe and effective .

Objective To investigate the expression of apolipoprotein M in rats after renal transplantation and its role and action mechanism in acute rejection (AR).Methods The kidney transplantation model in rats were established.Male SD and Wistar rats were used as donors,and Wistar rats as recipients.Three groups were designated:control group (syngeneic transplantation,n =24,recipients were treated with normal saline i.p.); AR group (allogeneic transplantation,n =24,recipients were treated with normal saline i.p.); PDTC group [allogeneic transplantation,n =24,recipients were treated with NF-κB inhibitor-pyrrolidine dithiocarbamat (PDTC) i.p.].The renal grafts were drawn at day 1,3,5 and 7 post-transplantation,and the expression levels of NF-κB P65 and apoM protein were detected by using Western blotting,and those of apoM,perforin and granzyme B mRNA were by using real-time PCR.The correlation of apoM and NF-κB,apoM and perforin,and apoM and granzyme B was respectively analyzed.Results As compared with control group,the expression levels of apoM,perforin and granzyme B mRNA in AR and PDTC groups were dramatically up-regulated at each time point (P＜0.01),and those in PDTC group were significantly lower than those in AR group (P＜0.01).The expression levels of apoM and NF-κB protein in AR group were both distinctly higher than those in control group (P＜0.01),and those in PDTC group were markedly lower than those in AR group (P＜0.01).A significantly positive correlation was found between the expression of apoM and NF-κB protein (r =0.469,P＜0.05).And the expression of apoM mRNA was positively correlated to perforin and granzyme B mRNA (r =0.731,P＜0.01 ; r =0.514,P＜0.05).Conclusion The expression of apoM is obviously up-regulated in renal grafts of rats,which may take part in the pathogenesis of AR via NF-κB.

Objective To detect serum biomarkers for pancreatic cancer associated diabetes and establish a model for diagnosis.Methods SELDI-TOF-MS was used to detect the differentially expressed serum proteins from 17 pancreatic cancer associated diabetes patients,17 new-onset type Ⅱ diabetes patients and 17 healthy controls,then a model of biomarkers was constructed and validated by Biomarker Patterns Software 5.0.Results Twelve discriminating m/z peaks were identified in the protein fingerprints in 10 pancreatic cancer associated diabetes patients,10 new-onset type Ⅱ diabetes patients and 10 healthy controls.Among them,the three biomarkers of mass/charge ratio 6116,6695 and 8936 were used to construct the model,which could diagnose 90％ pancreatic cancer associated diabetes form control groups.Blind test of other7 samples of three groups showed that 100％ pancreatic cancer associated diabetes,71％ new-onset diabetes and 86％ healthy controls were correctly classified.After searching protein database,there were metallothionein,pancreatic progenitor cell differentiation and proliferation factor-like protein,and fibroblastic growth factor 1,which were close to the weights of the above mentioned 3 differentially expressed proteins.Conclusions SELDI can identify 3 biomarkers for pancreatic cancer associated diabetes and a reliable model for diagnosis of pancreatic cancer associated diabetes is established.

Objective To investigate the apoptosis-inducing effect and anti-proliferative effect of epigallocatechin-3-gallate (EGCG) on human pancreatic cancer cell SW1990 in vitro. Methods The effect of proliferation was evaluated by MTT after the SW1990 cells in vitro were incubated with different concentrations of EGCG (6.25, 12.5, 25, 50, 100 μg/ml). The apoptosis-inducing effect was determined by flow cytometry after the cells were treated with 25 μg/ml of EGCG. The cell cycle of SW1990 cells was detected by flow cytometry after the cells incubated with different concentrations of EGCG (0, 10, 20, 30, 40, 50 μg/ml).Results After SW1990 cell were treated with different concentrations of EGCG (0, 25, 50 μg/ml), the values of A492 were 0.46 ±0.04,0.42 ±0.04,0.27 ±0.03 at 24 h; 0.48 ±0.02, 0.31 ±0.03,0.16 ±0.02at 48 h; 0.51 ±0.01,0.24 ±0.04,0. 14 ±0.04 at 72 h. EGCG inhibited the proliferation of SW1990 in a doseand time-dependant manner(P ＜0.01 ). The apoptotic rates at 24, 48, 72 h were (8.33 ± 1.15 )%, (19.77 ±0.81 )%, (29.17 ± 0.75 )% in the EGCG treatment group; while the corresponding values were (2.77 ±0.45 ) %, (3.20 ± 0.26 ) %, (3.67 ± 0.35 ) % in the control group; and the difference was statistically significant (P ＜0.01 ). After 0, 20, 50 μg/ml of EGCG treatment for 24 h, the percentages of SW1990 cellsin G0/G1 stage were (57.59 ±0.97)%, (62.99 ± 1.91 )%, (68.87 ± 1.88)%, and the percentages of SW1990 cells in G0/G1 stage increased with the increase of concentrations of EGCG, while the percentages of SW1990 cells in G2/M stage decreased with the increase of concentrations of EGCG (P ＜0.01 ). Conclusions EGCG can significantly inhibit the proliferation of SW1990 cells. The mechanism may be related to the apoptosis-inducing effect and the regulation of the cell cycle of the SW1990 cells.