Engineering of reaction specificity, enantioselectivity, and catalytic activity of nitrilase for highly efficient synthesis of pregabalin precursor.

Simultaneous evolution of multiple enzyme properties remains challenging in protein engineering. A chimeric nitrilase (BaNITM0 ) with high activity towards isobutylsuccinonitrile (IBSN) was previously constructed for biosynthesis of pregabalin precursor (S)-3-cyano-5-methylhexanoic acid ((S)-CMHA). However, BaNITM0 also catalyzed the hydration of IBSN to produce by-product (S)-3-cyano-5-methylhexanoic amide. To obtain industrial nitrilase with vintage performance, we carried out engineering of BaNITM0 for simultaneous evolution of reaction specificity, enantioselectivity, and catalytic activity. The best variant V82L/M127I/C237S (BaNITM2 ) displayed higher enantioselectivity (E = 515), increased enzyme activity (5.4-fold) and reduced amide formation (from 15.8% to 1.9%) compared with BaNITM0 . Structure analysis and molecular dynamics simulations indicated that mutation M127I and C237S restricted the movement of E66 in the catalytic triad, resulting in decreased amide formation. Mutation V82L was incorporated to induce the reconstruction of the substrate binding region in the enzyme catalytic pocket, engendering the improvement of stereoselectivity. Enantio- and regio-selective hydrolysis of 150 g/L IBSN using 1.5 g/L Escherichia coli cells harboring BaNITM2 as biocatalyst afforded (S)-CMHA with >99.0% ee and 45.9% conversion, which highlighted the robustness of BaNITM2 for efficient manufacturing of pregabalin.

Ginsenoside Re enhances the survival of H9c2 cardiac muscle cells through regulation of autophagy.

We examined the effect of ginsenoside Re (G-Re) on autophagy in H9c2 cardiomyocytes cultured in glucose deprivation (GD). Levels of the membrane-bound autophagy-related microtubule-associated protein 1A/1B-light chain 3 (LC3) B-2 were measured via immunoblotting and immunofluorescence was conducted to assess autophagosome formation. GD H9c2 cells were treated with 100 µmol/l G-Re. Cell viability was determined in culture medium. Phosphorylated 5' AMP-activated protein kinase (AMPK)-α and mammalian target of rapamycin (mTOR) levels were measured to explore the mechanisms underlying the effects of G-Re on autophagy in GD cells. G-Re treatment inhibited autophagosome formation and may be beneficial to GD cardiomyocytes.

[Study on quality standard of gypenosides extract and Gypenosides Tablets].

In response to no national standard for Gynostemma pentaphyllum, a market survey was carried out, and 17 batches of gypenosides extract and 29 batches of Gypenosides Tablets on the market were collected. With gypenoside A as an index, the TLC qualitative identification and HPLC quantitative evaluation method of gypenosides extract and tablets was established. Based on the determination results of 17 batches of gypenosides extract and 29 batches of Gypenosides Tablets, the quality standards of gypenosides extract and tablets were formulated respectively, so as to give suggestions for improving the quality standards of gypenosides extract and tablets. Compared with the existing ministerial standards, the qualitative identification and quantitative detection of specific components were added, in order to provide scientific basis and suggestions for the revision of the quality standard of gypenosides extract and tablet preparation.

[Comparative of functional components, antioxidant and α-glucosidase inhibition activities between Choerospondias axillaris fruit peel vinegar and apple vinegar].

Based on the idea of plant metabolomics, ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used to compare the chemical composition between 6 batches of fruit vinegar brewed from Choerospondias axillaris fruit peel and 6 batches of apple vinegar purchased from 3 companies. Antioxidant and α-glucosidase inhibition activities were also tested in vitro. A total of 43 compounds were identified by reference substance, liquid chromatography-mass spectrometry(LC-MS/MS) fragmentation information or literature data. A total of 40 compounds were identified in the C. axillaris fruit peel vinegar. A total of 16 compounds were identified in apple vinegar. There were 13 common ingredients including organic acids and esters such as citric acid, 2-isopropyl malic acid, and triethyl citrate. The results of partial leastsquares-discriminant analysis(PLS-DA) indicated that they had 33 significantly different compounds such as proanthocyanidin oligomer, quercetin-3-O-rhamnoside and heptadecanoic acid. The proanthocyanidins and flavonoid glycosides in C. axillaris peel vinegar were more abundant than apple vinegar, so it had better health function than ordinary fruit vinegar. The results showed that C. axillaris fruit peel vinegar had stronger antioxidant and α-glucosidase inhibition activities in vitro. The vinegar brewed from waste C. axillaris fruit peel had more chemical ingredients than the apple vinegar. C. axillaris fruit peel vinegar had better biological activity and health function, so it had good development prospect. This study provided the scientific evidence for exploiting the C. axillaris fruit peel into high value-added products. It also provided ideas for the comprehensive development and utilization of similar Chinese medicine waste.

[Visual analysis of coronavirus disease 2019 (COVID-19) studies based on bibliometrics].

To analyze the development of coronavirus disease 2019(COVID-19), this study systematically retrieved relevant Chinese and English literatures from both CNKI and Web of Science database platforms by bibliometric research method and CiteSpace 5.5.R2 software to obtain information and visualize relevant literatures. A total of 695 Chinese and 446 English literatures were included in this paper. Statistics showed that China had published most of the literatures and established close cooperation with the United States and the United Kingdom. Through the analysis, Tongji Medical College of Huazhong University of Science and Technology and its affiliated hospitals published the largest number of the publications. Moreover, the highly productive journals including Journal of Traditional Chinese Medicine and The Lancet covered eight major fields, such as medicine, medical virology, radiation medicine, infectious disease, and traditional Chinese medicine. Besides, a total of 35 special COVID-19 funds were recently established to subsidize these studies. The key words and themes analysis indicated that protein structure of COVID-19, receptor targets and mechanisms of action, integration of traditional Chinese and Western medicine, screening and development of antiviral drugs from traditional Chinese medicine and Western medicine, vaccine research as well as epidemiological characteristics and prediction are current study hotspots. This study provides a reference for researchers to rapidly master main study directions of COVID-19 and screen out relevant literatures.

[Comparison of Different Pretreatment Methods for DNA Extraction from Teeth].

OBJECTIVE: To compare the concentration of teeth DNA extracted by three different pretreatment methods and to explore a simple, economical and practical pretreatment method with high concentration of extracted DNA from teeth. METHODS: A total number of 21 molars were collected from 7 corpses. The pretreatment of 3 molars from each individual was randomly performed by tooth crumb method, ball-milling method and liquid nitrogen milling method and 50 mg tooth crumb was weight and DNA was extracted by AutoMate Express forensic DNA extraction system. Subsequently, the concentration of DNA and corresponding STR genotyping of three methods were compared. RESULTS: The DNA concentration extracted by tooth crumb method, ball-milling method and liquid nitrogen milling method was 0.055 6-1.989 1 ng/µL, 0.036 6-1.175 6 ng/µL and 0.037 8-1.249 0 ng/µL, respectively. The DNA concentration obtained by tooth crumb method was higher (P < 0.05) and the success rate of STR genotyping was high. CONCLUSION: Combined with AutoMate Express forensic DNA extraction system, tooth crumb method is an efficient and feasible method to extract DNA from teeth, which can be applied in forensic practice.

[Stability analysis of allelopathic effects of Panax notoginseng on main crops by AMMI model].

This paper is aimed to study the differences of allelopathic effects of Panax notoginseng under different allelopathic chemicals resources and selection of appropriate rotation crops. The additive main effects and multiplicative interaction ( AMMI) model had been used to evaluate the stability of allelopathic effects of P. notoginseng on the varieties of corn, wheat and rice properly. The model could use not only to evaluate the stability of non-regional trial data but also explore the interaction between the rotation crop genotypes and donor substances more efficiently. Meanwhile, correspondence analysis can be used in the AMMI to evaluate genotype stability and donor substances. Ejingza No. 1 (g6) had stronger allelopathic effects with high stability, but Yunrui No. 1 (g9) which was appropriate rotation crop genotype, had weaker allelopathic effects with high stability. These findings will aid in choosing appropriate rotation crops and establishing proper rotation system.

[Allelopathic effects of aqueous extracts from Panax notoginseng on three maize varieties (Zea mays)].

It has been showed that there were obvious obstacle effects of Panax notoginseng replanting. Crop rotation was the main effective technique to overcome the obstacle. To find a reasonable crop rotation system for P. notoginseng, aqueous extracts from root, stem and leaf of P. notoginseng were analyzed for allelopathic effect on three maize varieties (which are often grown in regions where P. notoginseng grown). The main results were as follows: (1) Allelopathic effect of P. notoginseng stem and leaf extracts on the three other tested plants was stronger than that of root extracts; (2) Corn was more vulnerable to the effects of allelochemicals at seedling stage than at germination stage, and the corn root was more sensitive than aerial part to allelochemicals; (3) Lusan No. 3 and Yunrui No. 1 showed resistance to P. notoginseng allelopathy, with respective comprehensive sensitivity indexes (M3) of - 0.089 3 and -0.159 2, while Bainuo No. 1 is sensitive at M3 = -0.261 0. It then can be concluded that Lusan No. 3 and Yunrui No. 1 may be an alternative rotation plants for overcoming P. notoginseng continuous cropping obstacle.

Dysfunctional circadian clock accelerates cancer metastasis by intestinal microbiota triggering accumulation of myeloid-derived suppressor cells.

Circadian homeostasis in mammals is a key intrinsic mechanism for responding to the external environment. However, the interplay between circadian rhythms and the tumor microenvironment (TME) and its influence on metastasis are still unclear. Here, in patients with colorectal cancer (CRC), disturbances of circadian rhythm and the accumulation of monocytes and granulocytes were closely related to metastasis. Moreover, dysregulation of circadian rhythm promoted lung metastasis of CRC by inducing the accumulation of myeloid-derived suppressor cells (MDSCs) and dysfunctional CD8+ T cells in the lungs of mice. Also, gut microbiota and its derived metabolite taurocholic acid (TCA) contributed to lung metastasis of CRC by triggering the accumulation of MDSCs in mice. Mechanistically, TCA promoted glycolysis of MDSCs epigenetically by enhancing mono-methylation of H3K4 of target genes and inhibited CHIP-mediated ubiquitination of PDL1. Our study links the biological clock with MDSCs in the TME through gut microbiota/metabolites in controlling the metastatic spread of CRC, uncovering a systemic mechanism for cancer metastasis.

Genetic polymorphism of 17 Y-STR loci in Han Chinese living in Lanzhou.

The genetic polymorphism across 17 Y-STR loci in a population of Han Chinese in Lanzhou was investigated. Haplotypes and allele frequencies for the 17 Y-chromosomal STRs loci DYS456, DYS389I, DYS390, DYS389II, DYS458, DYS19, DYS385a/b, DYS393, DYS391, DYS439, DYS635, DYS392, Y GATA H4, DYS437, DYS438 and DYS448 were determined in 500 healthy unrelated autochthonous males from Lanzhou. The results showed that no shared haplotypes were observed. Gene diversity values ranged from 0.3987 (DYS391) to 0.9740 (DYS385a,b). It was concluded that these loci will be very useful for human identification in forensic cases and paternity tests within the Han Chinese population inhabiting Lanzhou.

[Influence of diet behaviors on height among children and adolescents in China: a multiple level analysis].

OBJECTIVE: To categorize growth and development stages among children and adolescents based on height, and to explore the influences of diet behaviors on height during different growth and development stages. METHODS: Children and adolescents (7 to 18 years) with normal weights were selected using "Reference Norm for Screening Overweight and Obesity in Chinese Child and Adolescent" set up by WGOC in 2003 and "Reference Norm for Screening Underweight in Chinese Child and Adolescent" from the "2010 National Physical Fitness and Health Surveillance" data, and the variables of height and diet behaviors sorted. The growth and development stages were categorized using the hierachical cluster analysis, and the multilevel model was used to analyze influences of diet behaviors on height at different growth and development stages. RESULTS: Basis on height, there were 4 growth and development stages among the children and adolescents. In the boys, growth and development stages included Pre-GSS (growth spurt stage, GSS) including 7 to 10 years, GSS (11 to 12 years), Post-GSS (13 to 14 years), and growth stability stage (15 to 18 years); in the girls, the stages included the fast velocity GS (growth stage, GS) including 7 to 10 years, GS (11 to 12 years), Post-GS (13 to 15 years), and growth stability stage (16 to 18 years). The results of the multilevel model showed that the students' height in the urban areas were higher than in the rural areas (P<0.01), and the trend of difference between the urban and rural areas with the different growth stages was a parabola shape, the highest differences were 3.36 cm and 3.23 cm in the GSS and the fast velocity GS, respectively. There were significant influences of breakfast on height during the Pre-GSS and the fast velocity GS (P<0.01), and increased 0.40 cm and 0.57 cm, respectively. Excluding the stablility growth stage in the girls, drinking milk increased significantly height during the different growth stages (P<0.01), and the increases were gradual downtrend with the growth stages, the highest increase was 0.91 cm and 0.94 cm in Pre-GSS and the fast velocity GS, respectively. Eating eggs increased significantly height during all the growth stages (P<0.01), and the increases were the gradual uptrend with different growth stages. There were interaction effects among breakfast, drinking milk and eating eggs during the different growth stages, which was the gradual downtrend. CONCLUSION: Breakfast and drinking milk are conducive to growth during early adolescence, and the effect of eating eggs on growth is gradual uptrend with different growth stages.

[Association between early onset of menarche and anthropometry measurements among adolescent girls in China].

OBJECTIVE: To explore the association between early menarche with anthropometry measurements among adolescent girls in China. METHODS: Research material was selected from the data of 2010 Chinese National Surveys on Students Constitution and Health. Probability unit regression method was used to calculate the age of 10th percentile (P 10) at menarche and menarche age before the P 10 was defined as early menarche(9.0-11.6 years old). A total of 1072 girls with early menarche were screened. Each girl with early menarche was frequency matched with two girls who hadn't achieved menarche and with the age difference less than 0.1 yr and from the same urban or rural locations. A total of 2144 girls without menarche were screened. Participants' data of height, weight, sitting height, chest circumference, body mass index(BMI) and height and sitting height index were analyzed. t test and wilcoxon test were used to analyze the anthropometry measurements differences between the two groups, Chi square test was used to analyze the differences of overweight and obesity between the two groups. Multilevel model was used to explore the association between early menarche with anthropometry measurements and overweight and obesity. RESULTS: A total of 1072 girls with early menarche and 2144 girls without menarche. Early menarche girls' height, sitting height, chest circumference, weight and BMI were (151.42 ± 7.46) cm, (80.86 ± 4.21) cm, (73.88 ± 7.72) cm, (44.32 ± 9.35) kg, and (19.18 ± 3.03) kg/m(2), while they were (144.86 ± 7.55) cm, (76.96 ± 4.05) cm, (67.25 ± 6.94) cm, (36.07 ± 7.88) kg and (16.64 ± 2.48) kg/m(2) in girls without menarche. The difference between two groups were significant(Z values were -22.20, -23.69, -24.38, -23.12, -20.17, -6.33 respectively with all P values < 0.01). Multilevel analysis results showed that in 9.0-11.6 years old girls early menarche was associated with anthropometry measurements(all P values < 0.05). Compared with girls without menarche, girls with early menarche had a relative higher height, sitting height, weight, chest circumference, with increments of 5.28 cm, 3.37 cm, 5.53 cm, 6.37 kg, 1.79 kg/m(2). Within subgroup analysis, there were parabolic trends with age in the height, weight and sitting height differences and "U" trend in chest circumference difference between girls with early menarche and girls without menarche.In 9.0-10.8 years old, 10.9-11.0 years old and 11.1-11.2 years old groups, the risk of overweight and obesity among girls with early menarche were 2.98 (95%CI:1.92-4.63) times, 6.76 (95%CI:2.79-16.39) times, 2.99 (95%CI:1.40-6.40) times of girls without menarche. CONCLUSION: The early onset of menarche is related with height, sitting height, weight and chest circumference, and it is closely associated with overweight and obesity among adolescent girls in China.

Ginsenoside Rg1 inhibits autophagy in H9c2 cardiomyocytes exposed to hypoxia/reoxygenation.

Ginsenoside Rg1 promotes antioxidative protection and intracellular calcium homeostasis in cardiomyocytes hypoxia/reoxygenation (H/R) model. However, the pharmacological effects of G-Rg1 on autophagy in cardiomyocytes have not been reported. In this study, we employed H9c2 cardiomyocytes as a model to investigate the effects of G-Rg1 on autophagy in cardiomyocytes under H/R stress. Our results showed that H/R induced increased level of LC3B-2, an autophagy marker, in a time-dependent manner in association with decreased cell viability and cellular ATP content. H/R-induced autophagy and apoptosis were further confirmed by morphological examination. 100 µmol/l Rg1-inhibited H/R induced autophagy and apoptosis, and this was associated with the increase of cellular ATP content and the relief of oxidative stress in the cells. Mechanistically, we found that Rg1 inhibited the activation of AMPKα, promoted the activation of mTOR, and decreased the levels of LC3B-2 and Beclin-1. In conclusion, our data suggest that H/R induces autophagy in H9c2 cells leading to cell injury. Rg1 inhibits autophagosomal formation and apoptosis in the cells, which may be beneficial to the survival of cardiomyocytes under H/R.

Isolation and identification of the phenolic compounds from the roots of Sanguisorba officinalis L. and their antioxidant activities.

Four phenolic compounds were isolated from the roots of Sanguisorba officinalis L. by silica gel column chromatography and preparative HPLC. On the basis of chemical and spectroscopic methods, their structures were identified as methyl 4-O-ß-D-glucopyranosy-5-hydroxy-3-methoxylbenzoate (1), 3,3',4'-tri-O-methylellagic acid (2), fisetinidol-(4α-8)-catechin (3), and (+)-catechin (4). Compound 1 is a new phenolic glycoside and compounds 2 and 3 were isolated from the Sanguisorba genus for the first time. Compounds 1­4 were also assayed for their antioxidant activities using the DPPH free radical assay.

Terpene glycosides from the roots of Sanguisorba officinalis L. and their hemostatic activities.

Guided by a hemostasis bioassay, seven terpene glycosides were isolated from the roots of Sanguisorba officinalis L. by silica gel column chromatography and preparative HPLC. On the grounds of chemical and spectroscopic methods, their structures were identified as citronellol-1-O-α-L-arabinofuranosyl-(1→6)-ß-D-glucopyranoside (1), geraniol-1-O-α-L-arabinofuranosyl-(1→6)-ß-D-glucopyranoside (2), geraniol-1-O-α-Larabinopyranosyl-(1→6)-ß-D-glucopyranoside (3), 3ß-[(α-L-arabinopyranosyl)oxy]-19α-hydroxyolean-12-en-28-oic acid 28-ß-D-glucopyranoside (4), 3ß-[(α-L-arabinopyranosyl)-oxy]-19α-hydroxyurs-12-en-28-oic acid 28-ß-D-glucopyranoside (ziyu-glycoside I, 5), 3ß,19α-hydroxyolean-12-en-28-oic acid 28-ß-D-glucopyranoside (6) and 3ß,19α-dihydroxyurs-12-en-28-oic acid 28-ß-D-glucopyranoside (7). Compound 1 is a new mono-terpene glycoside and compounds 2, 3 and 5 were isolated from the Sanguisorba genus for the first time. Compounds 1­7 were assayed for their hemostatic activities with a Goat Anti-Human α2-plasmin inhibitor ELISA kit, and ziyu-glycoside I (5) showed the strongest hemostatic activity among the seven terpene glycosides. This is the first report that ziyu-glycoside Ι has strong hemostatic activity.

[Effects of overweight and obesity on physical function among children and adolescents: a multilevel analysis].

OBJECTIVE: To analyze the influence of overweight and obesity on body function among children and adolescents using a multilevel model, and to provide the basis for understanding their influence. METHODS: According to the "Reference Norms for Screening Overweight and Obesity in Chinese Children and Adolescents" set up by Working Group Obesity in China (WGOC) in 2003, we screened out individuals with overweight and obesity from the data obtained from the body and health survey of Chinese children and adolescents in 2005. The samples of the non-overweight group and the overweight group were randomly selected from the screened data by 1:1 match with samples of the obesity group by gender and city/rural area, then underwent the multilevel model to explore the influence of overweight and obesity on body function. RESULTS: The single dependent variable model showed that the overweight group had significant increases in systolic blood pressure (SBP) and diastolic blood pressure (DBP) by 5.25 mmHg (1 mmHg=0.133 kPa) and 3.44 mmHg, respectively (P<0.01), when compared with the non-overweight group. There were no significant differences between the overweight and non-overweight groups in pulse and vital capacity (VC) (P>0.05), and the obesity group had significant increases in SBP, DBP, pulse and VC by 10.15 mmHg, 6.34 mmHg, 1.44 times/min and 390.04 mL, respectively (P<0.01), when compared with the non-overweight group. The multiple dependent variables model showed that the SBP, DBP and pulse of the overweight group and obesity group were significantly greater than those of non-overweight group (P<0.01). There was a significant difference between the overweight group and non-overweight group in VC (P<0.05), yet the same result was not found between the obesity group and non-overweight group (P>0.05). CONCLUSION: Overweight and obesity may cause a decline in body function among children and adolescents, and obesity has a more obvious influence on body function.

Vitrectomy with residual internal limiting membrane covering and autologous blood for a secondary macular hole: A case report.

BACKGROUND: Myopic foveoschisis (MF) is a common complication of pathological myopia. A macular hole (MH) usually results from the natural progression of MF and is a common complication of vitrectomy. Vitrectomy combined with residual internal limiting membrane (ILM) covering and autologous blood was effective for closing a secondary MH. CASE SUMMARY: A 52-year-old woman presented to our clinic with a complaint of blurred vision in the right eye for 7 years. Her best corrected visual acuity (BCVA) was 20/100, axial length was 25.79 mm and standard equivalent refractive error was -10.5 dioptres. Preoperative optical coherence tomography revealed foveoschisis in the right eye. Vitrectomy with fovea-sparing ILM peeling was performed. An MH developed and gradually expanded 5 mo after the initial vitrectomy. Vitrectomy with residual ILM covering and autologous blood was performed. The MH closed 3 wk after the second vitrectomy. CONCLUSION: Fovea-sparing ILM peeling can provide residual ILM for the treatment of MH secondary to vitrectomy for MF. Vitrectomy combined with residual ILM covering and autologous blood is effective for closing secondary MH and improving BCVA.

A comprehensive review of traditional uses, phytochemistry and pharmacology of Reynoutria genus.

OBJECTIVES: The genus Reynoutria belonging to the family Polygonaceae is widely distributed in the north temperate zone and used in folk medicine. It is administered as a sedative, tonic and digestive, also as a treatment for canities and alopecia. Herein, we reported a review on traditional uses, phytochemistry and pharmacology reported from 1985 up to early 2022. All the information and studies concerning Reynoutria plants were summarized from the library and digital databases (e.g. ScienceDirect, SciFinder, Medline PubMed, Google Scholar, and CNKI). KEY FINDINGS: A total of 185 articles on the genus Reynoutria have been collected. The phytochemical investigations of Reynoutria species revealed the presence of more than 277 chemical components, including stilbenoids, quinones, flavonoids, phenylpropanoids, phospholipids, lactones, phenolics and phenolic acids. Moreover, the compounds isolated from the genus Reynoutria possess a wide spectrum of pharmacology such as anti-atherosclerosis, anti-inflammatory, antioxidative, anticancer, neuroprotective, anti-virus and heart protection. SUMMARY: In this paper, the traditional uses, phytochemistry and pharmacology of genus Reynoutria were reviewed. As a source of traditional folk medicine, the Reynoutria genus have high medicinal value and they are widely used in medicine. Therefore, we hope our review can help genus Reynoutria get better development and utilization.

[A case-control study of minimally invasive transforaminal interbody fusion with the assistance of robot and traditional fluoroscopy in the treatment of single-space lumbar disc herniation].

OBJECTIVE: To compare the safety and nail placement accuracy of fluoroscopy-assisted and robot-assisted minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) in the treatment of single-space lumbar disc herniation. METHODS: The clinical data of 52 patients with single-space lumbar disc herniation treated by MIS-TLIF from March 2019 to February 2020 were retrospectively analyzed. Among them, 24 patients were treated by robot-assisted MIS-TLIF(group A) and 28 patients were treated by fluoroscopy-assisted MIS-TLIF (group B). The intraoperative blood loss, operation time, intraoperative fluoroscopy times, preoperative and postoperative visual analogue scale(VAS), Japanese Orthopaedic Association(JOA) scores and operation-related complications were recorded in two groups. Gertzbein-Robbins grade according to CT scan was used to evaluate the nail placement after operation. Grade A and B were evaluated as satisfactory nail placement, and grade C, D, and E were evaluated as error placement. Babu's method was used to evaluate the screw's invasion to the superior articular process. RESULTS: The operation time, intraoperative blood loss and intraoperative fluoroscopy times in group A were less than those in group B(P<0.05).VAS and JOA scores of all patients at the final follow-up were significantly improved compared with those before operation(P<0.05), but there was no statistically significant difference between the groups(P>0.05). There were 96 and 112 screws in group A and group B, respectively. Three days after operation, according to the Gertzbein-Robbins grade to evaluate the nail placement accuracy, there were 90 screws of grade A, 5 of grade B, 1 of grade C, no grade D and E in group A;there were 84 screws of grade A, 16 of grade B, 8 of grade C, 4 of grade D, no grade E in group B;the difference between two groups was statistically significant(Z=-3.709, P=0.000). The satisfactory rate of screw placement in group A was 98.96% (95/96), and that of group B was 89.29% (100/112), the difference between two groups was statistically significant (χ2=8.254, P=0.004). Three days after operation, the invasion of superior facet joints by pedicle screws was evaluated according to Babu's method, including 90 screws in grade 0, 4 in grade 1, 2 in grade 2, and 0 in grade 3 in group A;86 in grade 0, 12 in grade 1, 10 in grade 2 and 4 in grade 3 in group B, and the difference was statistically significant(Z=-3.433, P=0.001). There were no serious spinal cord, nerve and vascular injuries and other operation-related complications caused by screw implantation failure in both groups. All patients were followed up from 6 to 12(9.06±1.60) months. The neurological symptoms improved well after operation. During the follow-up period, there was no recurrence of symptoms, loosening or breakage of the internal fixation. CONCLUSION: Compared with the traditional fluoroscopy-assisted MIS-TLIF, the spinal robot-assisted MIS-TLIF not only has more minimally invasive and safer, but also has higher accuracy in nail placement, lower incidence of upper articular process invasion, and more accurate decompression targets, which can be used for minimally invasive treatment of single-space lumbar disc herniation.

Blockade of adenosine A2A receptor alleviates cognitive dysfunction after chronic exposure to intermittent hypoxia in mice.

Obstructive sleep apnea-hypopnea syndrome (OSAHS) is widely known for its multiple systems damage, especially neurocognitive deficits in children. Since their discovery, adenosine A2A receptors (A2ARs) have been considered as key elements in signaling pathways mediating neurodegenerative diseases such as Huntington's and Alzheimer's, as well as cognitive function regulation. Herein, we investigated A2AR role in cognitive impairment induced by chronic intermittent hypoxia (CIH). Mice were exposed to CIH 7 h every day for 4 weeks, and intraperitoneally injected with A2AR agonist CGS21680 or A2AR antagonist SCH58261 half an hour before IH exposure daily. The 8-arm radial arm maze was utilized to assess spatial memory after CIH exposures.To validate findings using pharmacology, the impact of intermittent hypoxia was investigated in A2AR knockout mice. CIH-induced memory dysfunction was manifested by increased error rates in the radial arm maze test. The behavioral changes were associated with hippocampal pathology, neuronal apoptosis, and synaptic plasticity impairment. The stimulation of adenosine A2AR exacerbated memory impairment with more serious neuropathological damage, attenuated long-term potentiation (LTP), syntaxin down-regulation, and increased BDNF protein. Moreover, apoptosis-promoting protein cleaved caspase-3 was upregulated while anti-apoptotic protein Bcl-2 was downregulated. Consistent with these findings, A2AR inhibition with SCH58261 and A2AR deletion exhibited the opposite result. Overall, these findings suggest that A2AR plays a critical role in CIH-induced impairment of learning and memory by accelerating hippocampal neuronal apoptosis and reducing synaptic plasticity. Blockade of adenosine A2A receptor alleviates cognitive dysfunction after chronic exposure to intermittent hypoxia in mice.