BACKGROUND:Nobiletin has been found to improve lipopolysaccharide-induced abnormal activation of microglia,excessive release of inflammatory factors and redox imbalance.However,the specific mechanism is not fully understood. OBJECTIVE:To investigate the molecular mechanism by which nobiletin can inhibit lipopolysaccharide-induced inflammation in BV2 microglia. METHODS:Passage 3 BV2 microglia were divided into three groups:control group was cultured for 24 hours(without any treatment).Lipopolysaccharide group was treated with 10 μg/mL lipopolysaccharide for 24 hours.Lipopolysaccharide+nobiletin group was treated with 20 μmol/L nobiletin for 6 hours and then 10 μg/mL lipopolysaccharide for 24 hours.After the processing,cell proliferation was detected by CCK-8 assay.The level of intracellular reactive oxygen species was detected by fluorescent probe.The mRNA expression levels of nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β were detected by qRT-PCR.The protein expression levels of nuclear factor κB p65,p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β were detected by western blot assay. RESULTS AND CONCLUSION:(1)Compared with the control group,the proliferation activity of lipopolysaccharide group was decreased(P<0.001).Compared with the lipopolysaccharide group,the cell proliferation activity of lipopolysaccharide+nobiletin group was increased(P<0.001).(2)Compared with the control group,the level of intracellular reactive oxygen species was increased in the lipopolysaccharide group(P<0.001).Compared with the lipopolysaccharide group,the level of intracellular reactive oxygen species was decreased in the lipopolysaccharide+nobiletin group(P<0.01).(3)Compared with the control group,the mRNA expression levels of tumor necrosis factor α and interleukin-1β were increased in the lipopolysaccharide group(P<0.001,P<0.01).Compared with the lipopolysaccharide group,mRNA expression levels of tumor necrosis factor α and interleukin-1β were decreased in the lipopolysaccharide+nobiletin group(P<0.01,P<0.05).(4)Compared with the control group,the protein expression levels of p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β in were increased the lipopolysaccharide group(P<0.001).Compared with the lipopolysaccharide group,the expression of p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β was decreased in the lipopolysaccharide+nobiletin group(P<0.001).(5)These findings suggest that nobiletin attenuates lipopolysaccharide-induced inflammatory response in BV2 microglia by suppressing nuclear factor-κB signaling pathway.

Objective:To evaluate the structural and functional changes of the left atrium in amateur ultra-marathon runners using ultrasound three-dimensional speckle tracking imaging technology.Methods:From June 2023 to June 2024，90 amateur marathon athletes were retrospectively recruited，including 44 amateur ultra-marathon（running distance ≥50 km）athletes（Ultra-marathon，UM group）and 46 amateur classic marathon（running distance<50 km）athletes（Marathon，M group）. Another 39 healthy volunteers who underwent physical examinations in Hangzhou Normal University Affiliated Hospital at the same time were selected as the control group. The distance，running age of amateur marathon runners were recorded. All subjects which underwent conventional echocardiography combined with three-dimensional speckle tracking echocardiography（3D-STE）obtained left atrial volume and functional parameters，including two-dimensional ultrasound indicators such as left atrial anterior posterior diameter（LAd），left ventricular end-diastolic diameter（LVEDd），left ventricular ejection fraction（LVEF），early diastolic peak velocity at the mitral valve orifice（E），and early diastolic peak velocity at the lateral mitral annulus（e'），E/e'，left atrial minimum，maximum，presystolic volumes（LAVmin，LAVmax，LAVpreA），left atrial maximum volume index（LAVImax），left atrial emptying volume（LAEV），overall left atrial ejection fraction，active and passive ejection fraction（LAEF，LAaEF，LApEF），left atrial expansion index（LAEI），left atrial reservoir period，conduit period，systolic long axis and circumferential strain（LASr，LAScd，LASct，LASr-c，LAScd-c，LASct-c）. The differences in left atrial parameters among three groups were compared and the relevant factors affecting LAEF were analyzed.Results:There were statistically significant differences in LAd，LVEDd，LAVmin，LAVmax，LAVpreA，LAVImax，and LAEV among the three groups（all P<0.05），and there were statistically significant differences in pairwise comparisons between the groups（all P<0.05）. Compared with the control group，LAEF，LApEF，LASr，LAScd，LASct，LASr-c，LAScd-c and LASct-c were significantly reduced in the M and UM groups（all P<0.05），while there were no statistically significant differences in these parameters between the UM and M groups（all P>0.05）；There were no statistically significant differences in IVSd，LVEF，E/e'，LAEI，and LAaEF among the 3 groups（all P>0.05）.There were no statistically significant differences in IVSd，LVEF，E/e'，LAaEF and LAEI among the 3 groups（all P>0.05）. Multiple linear regression analysis showed that only weekly running volume was an independent predictor of decreased LAEF in amateur marathon runners. The repeatability test indicated good consistency within and between observers. Conclusions:Amateur marathon runners have varying degrees of damage to their left atrial reservoir function，conduit function，and contraction function，with amateur ultra-marathon runners showing more significant damages，3D-STE technology can evaluate the structural and functional changes in the left atrium in amateur ultra-marathon athletes，and assist clinical understanding of atrial adaptation in amateur ultra- marathon athletes.

Objective To investigate the clinical effect of far lateral approach surgery under unilateral biportal endoscope(UBE)for lumbar foraminal stenosis.Methods From June 2021 to August 2022,39 patients with lumbar foraminal stenosis were treated with far lateral approach surgery under UBE in our department.The patients were placed at the standard prone position,and a longitudinal line was drawn about 2 cm from the lateral edge of the upper and lower pedicles.Transverse incisions were made at the two intersection points of the horizontal line at the midpoint of the lateral edge of the upper and lower pedicles.After inserting the gradually enlarged sleeves to expand the space,the enlargement of the intervertebral foramen and release of the nerve root were completed under the direct view of UBE.The Visual Analogue Scale(VAS)of leg pain and the Oswestry Disability Index(ODI)were compared before and after operation.The effectiveness was evaluated according to the modified MacNab criteria at 12 months after operation.Results The operations were successfully completed in all the 39 cases.The operation time was(105.5±14.8)min,and the blood loss was(26.7±8.8)ml.The tear of adventitial nerve root occurred in 1 case during operation,and there was no nerve injury symptom after operation.One case of suspected infection occurred after operation and recovered after conservative treatment.All the 39 patients got out of bed with waist circumference on the first day after operation.The intervertebral foramen height measured by sagittal CT at 3 d after operation was(1.62±0.26)cm,which was significantly higher than that before operation[(1.51±0.28)cm;t=-4.010,P=0.000].The intervertebral foramen width was significantly greater than preoperation[(1.24±0.17)cm vs.(0.96±0.15)cm;t=-14.811,P=0.000].The area of intervertebral foramen was significantly larger than preoperation[(1.40±0.19)cm2 vs.(1.05±0.22)cm2;t=-12.874,P=0.000].The VAS of leg pain at 1 d,and 1,3,and 6 months after operation were significantly lower than those before operation(all P=0.000),and the VAS scores gradually decreased with the increase of time.The ODI was significantly lower at 1,3,and 6 months after operation than that before operation(all P=0.000),and the ODI decreased gradually with the increase of time.According to the modified MacNab criteria,at 12 months after operation,33 cases were excellent,4 cases were good,and 2 cases were fair,with an excellent and good rate of 94.9％(37/39).Conclusions Far lateral approach surgery under UBE for lumbar foraminal stenosis has significant effect on the relief of lumbago and leg pain and the improvement of lumbar function.It has little damage to lumbar stability and does not need internal fixation,which is worthy of clinical application and promotion.

Objective To investigate the clinical effect of far lateral approach surgery under unilateral biportal endoscope(UBE)for lumbar foraminal stenosis.Methods From June 2021 to August 2022,39 patients with lumbar foraminal stenosis were treated with far lateral approach surgery under UBE in our department.The patients were placed at the standard prone position,and a longitudinal line was drawn about 2 cm from the lateral edge of the upper and lower pedicles.Transverse incisions were made at the two intersection points of the horizontal line at the midpoint of the lateral edge of the upper and lower pedicles.After inserting the gradually enlarged sleeves to expand the space,the enlargement of the intervertebral foramen and release of the nerve root were completed under the direct view of UBE.The Visual Analogue Scale(VAS)of leg pain and the Oswestry Disability Index(ODI)were compared before and after operation.The effectiveness was evaluated according to the modified MacNab criteria at 12 months after operation.Results The operations were successfully completed in all the 39 cases.The operation time was(105.5±14.8)min,and the blood loss was(26.7±8.8)ml.The tear of adventitial nerve root occurred in 1 case during operation,and there was no nerve injury symptom after operation.One case of suspected infection occurred after operation and recovered after conservative treatment.All the 39 patients got out of bed with waist circumference on the first day after operation.The intervertebral foramen height measured by sagittal CT at 3 d after operation was(1.62±0.26)cm,which was significantly higher than that before operation[(1.51±0.28)cm;t=-4.010,P=0.000].The intervertebral foramen width was significantly greater than preoperation[(1.24±0.17)cm vs.(0.96±0.15)cm;t=-14.811,P=0.000].The area of intervertebral foramen was significantly larger than preoperation[(1.40±0.19)cm2 vs.(1.05±0.22)cm2;t=-12.874,P=0.000].The VAS of leg pain at 1 d,and 1,3,and 6 months after operation were significantly lower than those before operation(all P=0.000),and the VAS scores gradually decreased with the increase of time.The ODI was significantly lower at 1,3,and 6 months after operation than that before operation(all P=0.000),and the ODI decreased gradually with the increase of time.According to the modified MacNab criteria,at 12 months after operation,33 cases were excellent,4 cases were good,and 2 cases were fair,with an excellent and good rate of 94.9％(37/39).Conclusions Far lateral approach surgery under UBE for lumbar foraminal stenosis has significant effect on the relief of lumbago and leg pain and the improvement of lumbar function.It has little damage to lumbar stability and does not need internal fixation,which is worthy of clinical application and promotion.

Objective:To evaluate the structural and functional changes of the left atrium in amateur ultra-marathon runners using ultrasound three-dimensional speckle tracking imaging technology.Methods:From June 2023 to June 2024，90 amateur marathon athletes were retrospectively recruited，including 44 amateur ultra-marathon（running distance ≥50 km）athletes（Ultra-marathon，UM group）and 46 amateur classic marathon（running distance<50 km）athletes（Marathon，M group）. Another 39 healthy volunteers who underwent physical examinations in Hangzhou Normal University Affiliated Hospital at the same time were selected as the control group. The distance，running age of amateur marathon runners were recorded. All subjects which underwent conventional echocardiography combined with three-dimensional speckle tracking echocardiography（3D-STE）obtained left atrial volume and functional parameters，including two-dimensional ultrasound indicators such as left atrial anterior posterior diameter（LAd），left ventricular end-diastolic diameter（LVEDd），left ventricular ejection fraction（LVEF），early diastolic peak velocity at the mitral valve orifice（E），and early diastolic peak velocity at the lateral mitral annulus（e'），E/e'，left atrial minimum，maximum，presystolic volumes（LAVmin，LAVmax，LAVpreA），left atrial maximum volume index（LAVImax），left atrial emptying volume（LAEV），overall left atrial ejection fraction，active and passive ejection fraction（LAEF，LAaEF，LApEF），left atrial expansion index（LAEI），left atrial reservoir period，conduit period，systolic long axis and circumferential strain（LASr，LAScd，LASct，LASr-c，LAScd-c，LASct-c）. The differences in left atrial parameters among three groups were compared and the relevant factors affecting LAEF were analyzed.Results:There were statistically significant differences in LAd，LVEDd，LAVmin，LAVmax，LAVpreA，LAVImax，and LAEV among the three groups（all P<0.05），and there were statistically significant differences in pairwise comparisons between the groups（all P<0.05）. Compared with the control group，LAEF，LApEF，LASr，LAScd，LASct，LASr-c，LAScd-c and LASct-c were significantly reduced in the M and UM groups（all P<0.05），while there were no statistically significant differences in these parameters between the UM and M groups（all P>0.05）；There were no statistically significant differences in IVSd，LVEF，E/e'，LAEI，and LAaEF among the 3 groups（all P>0.05）.There were no statistically significant differences in IVSd，LVEF，E/e'，LAaEF and LAEI among the 3 groups（all P>0.05）. Multiple linear regression analysis showed that only weekly running volume was an independent predictor of decreased LAEF in amateur marathon runners. The repeatability test indicated good consistency within and between observers. Conclusions:Amateur marathon runners have varying degrees of damage to their left atrial reservoir function，conduit function，and contraction function，with amateur ultra-marathon runners showing more significant damages，3D-STE technology can evaluate the structural and functional changes in the left atrium in amateur ultra-marathon athletes，and assist clinical understanding of atrial adaptation in amateur ultra- marathon athletes.

BACKGROUND:Intervertebral disc degeneration is the basis of spinal degenerative diseases;however,there is no effective treatment. OBJECTIVE:To investigate whether sinomenine can inhibit interleukin-1β-induced apoptosis in nucleus pulposus cells and its molecular mechanism. METHODS:Rat nucleus pulposus cells were cultured in vitro by trypsin combined with type II collagenase digestion,and the cell growth curve was plotted.An appropriate sinomenine concentration was determined using the cell counting kit-8 kit.Nucleus pulposus cells were divided into control group,sinomenine group,interleukin-1β group,sinomenine+interleukin-1β group,zinc protoporphyrin group,zinc protoporphyrin+sinomenine group,zinc protoporphyrin+interleukin-1β group,and sinomenine+zinc protoporphyrin+interleukin-1β group.Proliferative activity,reactive oxygen species content,apoptosis rate,and heme oxygenase-1 expression in nucleus pulposus cells were detected. RESULTS AND CONCLUSION:The rat nucleus pulposus cells cultured in vitro were polygonal,triangular,and short wedge-shaped,and the cell growth showed an"S"curve.The cells grew slowly in the first 3 days of culture,rapidly in 4-6 days,and slowly again in 7-8 days.The cells then entered the"platform stage"where the number of cells no longer increased.The proliferative activity of myeloid cells showed no significant changes when the concentration of sinomenine was≤80 μmol/L(P>0.05).Interleukin-1β significantly reduced the proliferative activity of nucleus pulposus cells,increased the content of reactive oxygen species and led to apoptosis(P<0.01).Sinomenine intervention not only promoted heme oxygenase-1 expression(P<0.05)but also inhibited interleukin-1β-induced decrease in proliferative activity and increase in reactive oxygen species content and apoptosis rate in nucleus pulposus cells(P<0.05).These effects could be reversed by zinc protoporphyrin(P<0.01).

BACKGROUND:Intervertebral disc degeneration is caused by damage and degeneration of the nucleus pulposus and annulus fibrosus tissues inside the intervertebral disc,resulting in structural and functional changes of the intervertebral disc.However,there is yet no effective drug treatment for intervertebral disc degeneration. OBJECTIVE:To investigate the inhibitory effect of syringin on intervertebral disc degeneration. METHODS:A total of 10 male Sprague-Dawley rats were selected,and the coccygeal intervertebral disc(Co4/Co5)of each rat was set as model group,Co5/Co6 intervertebral disc as syringin group,and Co6/Co7 intervertebral disc as control group.The control group did not receive any treatment.In the model group and syringin group,a miniature puncture needle was used to puncture the annulus fibrosus to establish an intervertebral disc degeneration model.Immediately after modeling,2.5 μL of normal saline and syringin solution(5 μmol/L)were given in the model and syringin groups,respectively.Four weeks after injection,the samples were taken.The degree of intervertebral disc degeneration in rats was observed by hematoxylin-eosin and safranine O-fast green staining.The expressions of type Ⅱ collagen,aggrecan and matrix metalloproteinases 3 and 13 in intervertebral disc tissue were analyzed by immunohistochemical staining. RESULTS AND CONCLUSION:Hematoxylin-eosin staining showed that in the model group,the height of intervertebral disc decreased,the cartilage endplate became thinner and cracked,the fibrous ring structure was disordered and cracked,and the nucleus pulposus disappeared;in the syringin group,the height of intervertebral disc was normal or slightly lower than that in the control group,the degree of cartilage endplate degeneration was lighter than that in the model group,the fiber circle permutation was relatively regular with no cracks,and the nucleus pulposus was partially shrunk.Safranine O-fast green staining showed that in the model group,the cartilage endplate of the intervertebral disc was defective and the calcified layer of cartilage became thinner,showing obvious degeneration.The structure and morphology of intervertebral disc cartilage endplate in the syringin group recovered to some extent.Immunohistochemical staining showed that,compared with the control group,the expressions of type Ⅱ collagen and aggrecan in the intervertebral disc cartilage were decreased in the model group(P<0.000 1),while the expressions of matrix metalloproteinases 3 and 13 increased(P<0.000 1).Compared with the model group,the expressions of type Ⅱ collagen and aggrecan in the intervertebral disc cartilage tissue were increased in the syringin group(P<0.001,P<0.000 1),while the expressions of matrix metalloproteinases 3 and 13 decreased(P<0.001,P<0.000 1).These results showed that syringin could improve the structure and function of intervertebral disc by inhibiting the expression of matrix metalloproteinases 3 and 13 and increasing the expression of type Ⅱ collagen and aggrecan,thus preventing and slowing down the procession of intervertebral disc degeneration.

Objective:To assess the left ventricular(LV) structure and systolic function in amateur marathon runners using real-time three-dimensional speckle tracking echocardiography(3D-STE) and analyze its correlation with the running volume.Methods:A total of 84 amateur marathon runners were recruited between January 2019 and October 2021 in Hangzhou and were divided into short-term (ST) group(≤6 months) and more extended-term(MET) group(>6 months–2 years) based on their time of participating in the marathon. Thirty-nine healthy volunteers were enrolled from the Affiliated Hospital of Hangzhou Normal University during the same period as a control group. The running volume of ST and MET runners were recorded, LV end-diastolic volume(EDV), end-systolic volume(ESV), LV mass(LVM), LV ejection fraction(LVEF), LV global longitudinal strain(GLS), global circumferential strain(GCS), global radial strain(GRS), global area strain (GAS), twist, and torsion were measured by conventional echocardiography and 3D-STE. The differences of those parameters among the three groups were compared. The correlation between 3D-STE parameters and the running volume was further analyzed.Results:In ST amateur marathon runners, LV EDV was higher compared with controls( P<0.05). Compared with ST and control groups, LV EDV, ESV, and LVM of MET runners were increased(all P<0.05). In addition, compared with control group, GLS of MET runners was increased( P<0.05). The LVEF, GCS, GRS, GAS, twist, and torsion showed no statistically significant differences among the three groups(all P>0.05). In amateur marathon runners LV EDV, ESV, LVM had statistically significant positive correlations with an average weekly running volume and total running volume (all P<0.01). Multivariate linear regression analysis showed that the total running volume was an independent correlation factor in LV EDV of amateur marathon runners (β=0.618, P<0.01). Conclusions:Amateur marathon runners participating in a short term marathon mainly show an increase in LV EDV, the longitudinal systolic function of the LV can be enhanced in the early stage of the marathon.

Objective:To assess the left ventricular (LV) systolic function in amateur marathoners by two-dimensional speckle tracking echocardiography.Methods:A total of 59 amateur marathon runners were recruited from January 2019 to June 2020 in Hangzhou and were divided into group A (>2-5 years) and group B (>5 years) based on their time of participating in marathon. Thirty-one healthy volunteers were enrolled from Affiliate Hospital of Hangzhou Normal University during the same period as a control group. Conventional echocardiography combined with two-dimensional speckle tracking imaging were applied to all the subjects to obtain interventricular septum diastolic thickness (IVSd), LV posterior wall thickness(PWd), LV end-diastolic diameter (LVEDd), relative wall thickness(RWT) and LV mass(LVM), LV end-diastolic volume(EDV), LV end-systolic volume(ESV) and stroke volume(SV), LV ejection fraction(LVEF), LV global longitudinal strain (GLS), and global circumferential strain (GCS). Pre-marathon, 1 hour and 4th day post-marathon echocardiography were performed in amateur marathon runners.Results:Compared with the control group, group A amateur marathon runners showed significant increases in IVSd, PWd, LVEDd, RWT and LVM(all P<0.05); In addition, the IVSd, PWd, LVEDd, RWT, LVM, EDV, ESV and SV in group B runners were further increased compared to those of group A runners (all P<0.01). The LVEF and pre-marathon GCS showed no statistically significant differences among the three groups (all P>0.05), while the pre-marathon GLS showed a statistically significant difference among the three groups ( P<0.01). Compared with group A runners, the pre-marathon, 1 hour and 4th day post-marathon GLS and 1 hour post-marathon GCS were significantly decreased in group B runners (all P<0.01); In intra-group comparison, 1 hour post-marathon GLS was significantly decreased in relative to pre-marathon and 4th day post-marathon GLS in both group A and B (all P<0.05). One hour post-marathon GCS was significantly decreased compared with pre-marathon GCS in group B ( P<0.05). Conclusions:Amateur runners who have participated in long time marathon have reduced LV longitudinal strains and transient post-marathon decreases in LV systolic function.

BACKGROUND:Intervertebral disc degeneration is the pathological basis of degenerative spinal diseases. Studies on the influentialfactors of intervertebral disc degeneration contribute to the prevention and treatment of degenerative spinal disease.
OBJECTIVE:To observe the growth and proliferation of nucleus pulposus cels isolated by trypsin plus type II colagenase digestion in complete medium with different glucose concentrations, exploring the optimal glucose concentration for growth of nucleus pulposus cels.
METHODS:Nucleus pulposus cels isolated and cultured by trypsin plus type II colagenase digestion method were observed under an inverted microscope, and thecelnumber was counted. Morphology of nucleus pulposus cels was observed afterhematoxylin-eosinstaining and toluidine blue staining. Colagen type II immunoreactivity was detected by immunohistochemical staining combined with immunofluorescent staining.Nucleus pulposuscels were incubated in complete medium containing various glucose concentrations (0, 6.25, 12.5, 17.5, and 25 mmol/L) for 24 hours, and then cel proliferation and apoptosis were determined.
RESULTS AND CONCLUSION:The stained nucleus pulposus cels showed polygonal and short spindle, with one or two nuclei. Celularpseudopod appeared gradualy and then became slim with increased passage numbers. The isolated and cultured nucleus pulposus cels positively expressed colagen type II and aggrecanProliferative activity of nucleus pulposus cels cultured in medium with 17.5 mmol/L glucose was significantly higher than that in medium with 0 and 25 mmol/L glucose (P< 0.05 orP< 0.01). There wasno significant differencein cel apoptosis between these groups except for 0 mmol/L glucose (P<0.05). These results confirm that a large number of nucleus pulposus celscan beharvested by trypsin plustype II colagenase digestion and the optimal glucose concentration is 17.5 mmol/L.