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.

Objective:To investigate the effects of self-made Gujin Xiaoji Mixture combined with warming needle therapy on the clinical efficacy and immune function of patients with advanced non-small cell lung cancer (NSCLC) with qi and yin deficiency syndrome.Methods:This experiment was a randomized controlled trial study. 180 patients with advanced NSCLC qi and yin deficiency syndrome in the oncology centre of Qingdao Hospital of Traditional Chinese Medicine were selected as the observation subjects from March 2021 to August 2022, and were divided into 2 groups using the random number table method, with 90 cases in each group. The control group received conventional chemotherapy combined with Sintilimab injection, 21 days as a cycle, with a total of 4 cycles of treatment; and the observation group received Gujin Xiaoji Mixture combined with warming needle therapy based on the control group, 7 days as one course of treatment, with a total of 12 courses. Both groups were followed up for 12 months. The TCM syndrome scores were performed before and after treatment. The functional assessment of cancer therapy-lung (FACT-L) was used to evaluate the quality of life of patients; flow cytometry was used to detect the levels of CD3 +, CD4 +, CD8 + and NK cell, and the CD4 +/CD8 + ratio was calculated; adverse drug reactions and progression free survival of patients during treatment were observed and recorded, the efficacy of TCM syndrome and objective efficacy of solid tumors were evaluated. Results:After treatment, the observation group's post-treatment TCM syndrome score (5.67±1.99 vs. 7.12±2.31, t=-4.53) was lower than that of the control group ( P<0.001); mobility (23.03±2.80 vs. 20.69±2.46, t=5.96), daily living (23.06±2.56 vs. 20.71± 2.33, t=6.42), emotional status (18.44±2.32 vs. 16.12±2.71, t=6.18), and other factors (33.14±4.11 vs. 27.39±4.64, t=8.81) and total score (97.68±7.23 vs. 84.91±7.49, t=11.64) were higher than those in the control group ( P<0.01). In the observation group, after treatment, the levels of CD3 + [(65.14±6.06)% vs. (59.84±5.74)%, t=6.02], CD4 + [(40.09±4.09)% vs. (35.69±3.86)%, t=7.43], NK cell [(29.11±4.81)% vs. (22.38±4.51)%, t=9.68] and CD4 +/CD8 + [(1.52±0.27) vs. (1.14±0.12), t=12.63] were higher than those in the control group ( P<0.01), and CD8 + [(26.82±3.79)% vs. (31.76±4.65)%, t=-7.81] level was lower than that of the control group ( P<0.01). After treatment, the objective remission rate in the observation group was 7.8% (7/90), and the disease control rate was 87.8% (79/90), while the objective remission rate after treatment in the control group was 5.5% (5/90), and the disease control rate was 82.2% (74/90), and there were no statistical significance in the comparison of objective remission rate and disease control rate of the 2 groups ( χ2=0.09, 0.70, P=0.765, 0.407). The total effective rate after treatment was 62.2% (56/90) in the observation group and 34.4% (31/90) in the control group, and the difference between the 2 groups was statistically significant ( Z=-3.89, P<0.001). WBC [(4.27±1.12)×10 9/L vs. (3.84±1.11)×10 9/L, t=2.58] and haemoglobin [(119.93±17.25)g/L vs. (109.76±15.61)g/L, t=4.15] levels of the observation group were higher than those in the control group after treatment ( P<0.01). During follow-up, the median progression-free survival was 6.2 months in the observation group and 5.5 months in the control group patients, and the difference between the 2 groups was not statistically different ( t=0.11, P>0.05). Conclusion:The combination of Gujin Xiaoji Mixture with warming needle therapy can effectively improve the clinical symptoms of patients with advanced NSCLC with deficiency of qi and yin syndrome, improve the immunity and clinical efficacy of patients, alleviate the adverse effects of drugs, and prolong the progression-free survival period.

Bone organoids can simulate the complex structure and function of the bone tissues, which makes them a frontier technology in organoid researches. Bone organoids show a tremendous potential of applications in bone disease modeling, bone injury repair, and medicine screening. Although advancements have been made so far in constructing bone organoids with functional structures like mineralization, bone marrow, trabecular bone, callus, woven bone, etc, the researches in this field are confronted with numerous challenges such as lack of standardized construction strategies and unified evaluation criteria, which limits their further promotion and application. To standardize researches in bone organoids, the Orthopedic Expert Committee of Geriatric Branch of Chinese Association of Gerontology and Geriatrics, the Youth Osteoporosis Group of Orthopedic Branch of Chinese Medical Association, the Osteoporosis Group of Orthopedic Surgeon Branch of Chinese Medical Doctor Association, and the Osteoporosis Committee of Shanghai Association of Integrated Traditional Chinese and Western Medicine organized related experts to formulate Expert consensus on the construction, evaluation, and application of bone organoids ( version 2024) based on an evidence-based approach. A total of 17 recommendations were put forth, aiming to standardize researches and clinical applications of bone organoids and enhance their value in scientific research and clinical practice.

Objective:To investigate the efficacy and safety of anlotinib combined with gemcitabine + cisplatin (GP) regimen in the treatment of patients with recurrent or metastatic nasopharyngeal carcinoma.Methods:A retrospective case controlled study was conducted. The clinical data of 82 patients with nasopharyngeal carcinoma newly diagnosed as distant metastasis or recurrence and metastasis after radical chemoradiotherapy in the Second Affiliated Hospital of Guangxi University of Science and Technology and Laibin People's Hospital from January 2020 to December 2021 were retrospectively analyzed. Among 82 patients, 45 patients receiving anlotinib combined with GP regimen were treated as the treatment group, and 37 patients receiving GP chemotherapy regimen during the same period were treated as the control group. Short-term efficacy and adverse reactions were compared between the 2 groups. Kaplan-Meier method was used to compare the progression-free survival (PFS) and overall survival (OS), and log-rank test was performed. Cox proportional risk model was used to analyze the factors influencing the PFS of patients.Results:Among 82 patients, 63 were males and 19 were females, aged (48±10) years. The objective response rate (ORR) and disease control rate (DCR) in the treatment group were higher than those in the control group [ORR: 71.11% (32/45) vs. 62.16% (23/37); DCR: 86.67% (39/45) vs. 81.08% (30/37)], while there were no statistically significant differences (all P > 0.05). The median PFS time was 23.53 months (95% CI: 17.48-29.58 months), 17.40 months (95% CI: 13.33-21.47 months), respectively in the treatment group and the control group, and the difference in PFS was statistically significant ( P < 0.05). The median OS time was 34.03 months (95% CI: 29.42-38.64 months), 30.47 months (95% CI: 28.28-32.66 months), respectively in the treatment group and the control group, and the difference in OS was not statistically significant ( P > 0.05). Oral anlotinib, recurrence or metastasis at initial diagnosis were independent factors influencing the PFS of patients. The main adverse effects of anrotinib were grade 1-2 anorexia, fatigue, granulocytopenia, hypertension, hand-foot syndrome, oral mucositis, and liver function damage. The incidence of grade 3-4 was low, and the adverse reactions in most patients could be controlled after symptomatic treatment and adjustment of drug dosage. Conclusions:For patients with recurrent or metastatic nasopharyngeal carcinoma, oral anlotinib targeted therapy combined with GP chemotherapy regimen can prolong the PFS time of patients. Most anlotinib-related side effects can be tolerated.Objective To investigate the efficacy and safety of anlotinib combined with gemcitabine + cisplatin (GP) regimen in the treatment of patients with recurrent or metastatic nasopharyngeal carcinoma. Methods A retrospective case controlled study was conducted. The clinical data of 82 patients with nasopharyngeal carcinoma newly diagnosed as distant metastasis or recurrence and metastasis after radical chemoradiotherapy in the Second Affiliated Hospital of Guangxi University of Science and Technology and Laibin People's Hospital from January 2020 to December 2021 were retrospectively analyzed. Among 82 patients, 45 patients receiving anlotinib combined with GP regimen were treated as the treatment group, and 37 patients receiving GP chemotherapy regimen during the same period were treated as the control group. Short-term efficacy and adverse reactions were compared between the 2 groups. Kaplan-Meier method was used to compare the progression-free survival (PFS) and overall survival (OS), and log-rank test was performed. Cox proportional risk model was used to analyze the factors influencing the PFS of patients. Results Among 82 patients, 63 were males and 19 were females, aged (48±10) years. The objective response rate (ORR) and disease control rate (DCR) in the treatment group were higher than those in the control group [ORR: 71.11% (32/45) vs. 62.16% (23/37); DCR: 86.67% (39/45) vs. 81.08% (30/37)], while there were no statistically significant differences (all P > 0.05). The median PFS time was 23.53 months (95% CI: 17.48-29.58 months), 17.40 months (95% CI: 13.33-21.47 months), respectively in the treatment group and the control group, and the difference in PFS was statistically significant ( P < 0.05). The median OS time was 34.03 months (95% CI: 29.42-38.64 months), 30.47 months (95% CI: 28.28-32.66 months), respectively in the treatment group and the control group, and the difference in OS was not statistically significant ( P > 0.05). Oral anlotinib, recurrence or metastasis at initial diagnosis were independent factors influencing the PFS of patients. The main adverse effects of anrotinib were grade 1-2 anorexia, fatigue, granulocytopenia, hypertension, hand-foot syndrome, oral mucositis, and liver function damage. The incidence of grade 3-4 was low, and the adverse reactions in most patients could be controlled after symptomatic treatment and adjustment of drug dosage. Conclusions For patients with recurrent or metastatic nasopharyngeal carcinoma, oral anlotinib targeted therapy combined with GP chemotherapy regimen can prolong the PFS time of patients. Most anlotinib-related side effects can be tolerated.

Objective To investigate the value of a nomogram model constructed from intratumoral and peritumoral magnetic resonance imaging radiomics combined with clinical characteristics in predicting the status of lymphovascular space invasion(LVSI)in cervical cancer.Methods A retrospective analysis was conducted on 178 cervical cancer patients confirmed by postoperative pathology,with 70 cases of LVSI(+)and 108 cases of LVSI(-).The patients were divided into a training set[142 cases,including 54 cases of LVSI(+)and 88 cases of LVSI(-)]and a test set[36 cases,including 16 cases of LVSI(+)and 20 cases of LVSI(-)]at a ratio of 8:2.All underwent magnetic resonance imaging before surgery,and regions of interest were manually delineated layer by layer on the T2WI sequence,with the peritumoral region being uniformly expanded outward.Univariate logistic analysis was performed on clinical factors to select independent factors for cervical cancer LVSI(+).Radiomic features were extracted separately from the intratumoral region,the peritumoral region,and the intratumoral-peritumoral region to construct radiomics models,and the differences between the peritumoral and the intratumoral-peritumoral models were compared.A combined model was established based on the radiomics scores of the optimal intratumoral-peritumoral model and clinical independent predictive factors,and a nomogram was plotted.Receiver operating characteristic curves were used to evaluate the diagnostic performance of each model,and decision curve analysis was used to assess the clinical value of the models.Results The combined model demonstrated the best performance among the various models,with AUC of 0.970 in the training set and 0.803 in the test set.Conclusion Intratumoral and peritumoral magnetic resonance imaging radiomics combined with clinical characteristics can effectively predict LVSI in cervical cancer.

Objective:To investigate college students' awareness of dry eye and the prevalence of symptomatic dry eye disease.Methods:From October 2019 to January 2020, self-designed questionnaires including basic information, daily habits, dry eye cognition, and the Ocular Surface Disease Index (OSDI) scale, were distributed to students at Sichuan University. Univariate analysis and multivariate analysis were performed using SPSS 25.0 software.Results:A total of 1 013 valid questionnaires were collected. The awareness rate of dry eye among college students was 54.9% (556/1 013). The average OSDI scale score of college students was 14.02 ± 12.35 points. A total of 501 (49.5%) college students were judged to develop symptomatic dry eye disease by OSDI scale screening, of which 301 (29.7%) college students had mild dry eye disease, 124 (12.2%) college students had moderate dry eye disease and 76 (7.5%) college students had severe dry eye disease. Binary logistic regression analysis showed that the number of adverse habits associated with dry eyes ( P = 0.003), sleep quality at night ( P < 0.001), use of over-the-counter eye drops ( P < 0.001), and daily time spent in reading video terminals ( P < 0.001) were the influential factors of symptomatic dry eye disease screened by OSDI. Conclusion:College Students' awareness of dry eye needs to be improved. The prevalence of symptomatic dry eye disease in college students judged by the OSDI scale is high.

Objective:To summarize the clinical feature of functional distant metastasis (DM) of differentiated thyroid cancer (DTC) and observe the efficacy of 131I treatment. Methods:Between August 2008 and January 2021, a total of 13 DTC patients (4 males, 9 females; age 26-75 years) with functional DM from Henan Provincial People′s Hospital were retrospectively analyzed. Clinical data of patients were collected, including pathological type, metastasis size, metastasis location, and thyroid stimulating hormone (TSH) before the first 131I treatment. Efficacy of 131I treatment in patients with functional DM-DTC was evaluated by response evaluation criteria in solid tumors (RECIST) 1.1 and thyroglobulin (Tg). Complete remission (CR) and partial remission (PR) were considered as effective. Wilcoxon signed rank test was used to analyze the maximum diameter change of metastatic lesions before and after 131I treatment. Results:Among 13 DM-DTC patients, 8 were follicular thyroid cancer (FTC), 5 were papillary thyroid cancer (PTC). Metastasis lesions were mainly located in lungs ( n=12) and bones ( n=6). There were 12 patients with maximum metastasis diameter ≥1 cm, and 3 patients with TSH≥30 mU/L before the first 131I treatment. Nine patients were assessed as PR by RECIST 1.1, 3 patients were assessed as CR by RECIST 1.1 and the value of Tg, and 1 patient was assessed as PR by the changing of Tg. The effective rate of 131I treatment for patients with functional DM-DTC was 13/13. The maximum metastasis diameter was significantly decreased after 131I treatment (2.6(1.6, 3.3) vs 1.2(0.1, 2.2) cm; z=-3.06, P=0.002). Conclusion:Patients with functional DM-DTC are characterized by high proportion of FTC and the maximum metastasis diameter ≥1 cm, low proportion of TSH ≥30 mU/L before the first 131I treatment, and high effective rate of 131I treatment.

Non-conventional yeasts such as Yarrowia lipolytica, Pichia pastoris, Kluyveromyces marxianus, Rhodosporidium toruloides and Hansenula polymorpha have proven to be efficient cell factories in producing a variety of natural products due to their wide substrate utilization spectrum, strong tolerance to environmental stresses and other merits. With the development of synthetic biology and gene editing technology, metabolic engineering tools and strategies for non-conventional yeasts are expanding. This review introduces the physiological characteristics, tool development and current application of several representative non-conventional yeasts, and summarizes the metabolic engineering strategies commonly used in the improvement of natural products biosynthesis. We also discuss the strengths and weaknesses of non-conventional yeasts as natural products cell factories at current stage, and prospects future research and development trends.
Yeasts/genetics* ; Yarrowia/metabolism* ; Gene Editing ; Metabolic Engineering

ObjectiveTo investigate the protective effect of modified Huangqi Guizhi Wuwutang （MHGW） on endoplasmic reticulum stress in the sciatic nerve of diabetes rats based on the pathways of inositol-requiring enzyme 1α （IRE1α） and CCAAT/enhancer-binding protein homologous protein （CHOP）. MethodSixty rats were fed on a high-sugar and high-fat diet for six weeks， followed by intraperitoneal injection of streptozotocin at a dose of 35 mg·kg^-1. Random blood glucose levels were measured three days later and rats with a sustained blood glucose level ≥ 16.7 mmol·L^-1 were included in study （n=48）. The rats were randomly divided into a model group， an α-lipoic acid group （0.026 8 g·kg^-1·d^-1）， a high-dose MHGW group （2.5 g·kg^-1·d^-1）， and a low-dose MHGW group （1.25 g·kg^-1·d^-1）. Another 10 rats were assigned to the normal group. The intervention lasted for 16 weeks. After 16 weeks， the sciatic nerve structure of the rats in each group was observed under light microscopy using Luxol fast blue （LFB） staining. Transmission electron microscopy was used to observe the ultrastructure of the sciatic nerve. Chemiluminescence method was employed to measure the serum reactive oxygen species （ROS） levels. Western blot and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to evaluate the expression of p-IRE1α protein， IRE1α mRNA， CHOP protein， and CHOP mRNA in the sciatic nerve of the rats. ResultCompared with the normal group， the model group showed elevated serum ROS levels （P<0.01）. In contrast， the serum ROS levels were significantly reduced in the treatment groups compared with those in the model group （P<0.01）. The sciatic nerve of the model group showed pathological changes compared with that in the normal group， while the treatment groups exhibited improvement in sciatic nerve pathology compared with the model group. The protein expression of p-IRE1α and CHOP in the sciatic nerve significantly increased in the model group as compared with that in the normal group （P<0.01）. However， the treatment groups showed a significant decrease in the protein expression of p-IRE1α and CHOP in the sciatic nerve compared with the model group （P<0.05， P<0.01）. Furthermore， compared with the normal group， the model group showed upregulated mRNA expression of IRE1α and CHOP in the sciatic nerve （P<0.01）， while the treatment groups exhibited a significant decrease in the mRNA expression of IRE1α and CHOP compared with the model group （P<0.01）. ConclusionMHGW can alleviate endoplasmic reticulum stress-induced cell apoptosis and improve the structure and function of the sciatic nerve in diabetes rats by inhibiting the expression of IRE1α/CHOP pathway-related proteins and mRNA， thereby preventing and treating peripheral neuropathy in diabetes.

ObjectiveTo investigate the effect of modified Huangqi Guizhi Wuwutang （MHGW） on the protein and mRNA expression of B-cell lymphoma-2-associated X protein （Bax） and cysteinyl aspartate specific proteinase-12 （Caspase-12） related to the apoptosis of sciatic nerve cells in diabetes rats to explore the mechanism of MHGW in the treatment of peripheral neuropathy in diabetes. MethodAnimal experiments were conducted. A diabetes model was induced in sixty male sprague-dawley （SD） rats by feeding on a high-sugar and high-fat diet combined with streptozotocin （STZ） intraperitoneal injection. Rats with random blood glucose levels ≥ 16.7 mmol·L^-1 for three consecutive days were considered to have successfully developed diabetes. Forty-eight rats that successfully developed diabetes were randomly divided into a model group， an α-lipoic acid group （0.026 8 g·kg^-1·d^-1）， a high-dose MHGW group （2.5 g·kg^-1·d^-1）， and a low-dose MHGW group （1.25 g·kg^-1·d^-1）， with 12 rats in each group. Another 10 rats were assigned to the normal group. Body weight and random blood glucose levels of the rats were monitored. At the end of a 16-week intervention period， the sciatic nerve conduction velocity of the rats was measured using the Key point electromyography collection system. The protein and mRNA expression of Bax and Caspase-12 in the sciatic nerve cells was detected by Western blot analysis and real-time quantitative polymerase chain reaction （Real-time PCR）， respectively. ResultCompared with the normal group， the model group showed a significant decrease in body weight （P<0.01） and a significant increase in random blood glucose levels （P<0.01）. After a 16-week intervention， compared with the model group， the high-dose MHGW group exhibited a significant increase in body weight （P<0.05）， while there were no statistically significant differences in body weight changes among the other treatment groups. Random blood glucose levels significantly decreased in all treatment groups （P<0.01）. After 16 weeks of intervention， compared with the normal group， the model group had significantly reduced motor and sensory nerve conduction velocities （P<0.01）. Compared with the model group， all treatment groups showed significant increases in motor and sensory nerve conduction velocities （P<0.05， P<0.01）. The expression of Bax and Caspase-12 proteins in the sciatic nerve cells was significantly elevated in the model group compared with that in the normal group （P<0.01）. In contrast， all treatment groups showed significant reductions in the expression of Bax and Caspase-12 proteins in the sciatic nerve cells as compared with that in the model group （P<0.01）. The expression of Bax and Caspase-12 mRNA in the sciatic nerve cells significantly increased in the model group compared with that in the normal group （P<0.01）. Compared with the model group， the α-lipoic acid group and the high-dose MHGW group showed significant reductions in the expression of Bax mRNA in the sciatic nerve cells （P<0.05， P<0.01）， while the low-dose MHGW group showed a decreasing trend in the expression of Bax mRNA. The expression of Caspase-12 mRNA in the sciatic nerve cells significantly decreased in all treatment groups （P<0.01）. ConclusionMHGW may improve and repair sciatic nerve damage in diabetes rats by inhibiting sciatic nerve cell apoptosis.