Objectives:To explore the impact of Gli1(gioma-associated oncogene homolog)on nucleus pulposus cells(NPCs)and its role in the fibrotic changes of the nucleus within intervertebral discs.Methods:A rat tail intervertebral disc puncture model was established.Histological staining was used to evaluate the levels of intervertebral disc degeneration and fibrotic changes in the untreated(negative control,NC)group and the puncture(puncture-induced intervertebral disc degeneration,PIDD)group.Western blot was used to assess the expression levels of Gli1 protein in the tissues.Gli1 protein was engineered to overexpress by lentivirus in NPCs and verified.The experiment was divided into three groups:control(Blank),Gli1 overexpression(Lv-Gli1),and virus control(lv-Ctrl).Immunofluorescence and Western blot were used to detect the expression levels of fibrotic markers FAP and FSP-1.Adding Gli1 inhibitor GANT61,cellular experiment was grouped into control(blank),Gli1 overexpression(Lv-Gli1),Gli1 overexpression+GANT61(Lv-Gli1+GANT61),and Gli1 overexpression+DMSO(Lv-Gli1+DMSO).Western blot was used to detect the expression levels of FAP and FSP-1 proteins.In vivo animal experiments were conducted in groups of puncture(PIDD),puncture treatment(PIDD+GANT61),and treatment control(PIDD+DMSO).Histological staining was used to evaluate the levels of intervertebral disc degeneration and fibrotic changes.Results:Histological staining results showed the histological score of degenerated discs significantly increased(P＜0.01).Western blot results showed a significant increase in Gli1 protein expression in the degenerated group(P＜0.05),which was consistent with the conclusions of the cellular experiments.NPCs overexpressing Gli1 protein were constructed,and immunofluorescence staining results showed that the expression of FAP and FSP-1 in the Gli1 overexpression group was significantly upregulated compared to the control and virus control groups.Western blot results further showed a significant increase in the expression of fibrotic markers in the Gli1 overexpression group(P＜0.05).In the Gli1 overexpression+GANT61 group of the cellular experiment,compared to the control and Gli1 overexpression groups,Western blot indicated that the expressions of FAP and FSP-1 proteins were downregulated(P＜0.05).In the in vivo experiment,the PIDD+GANT61 group showed a significant improvement in histological scores and collagen composition(P＜0.05).Conclusions:The expression of Gli1 protein is significantly upregulated in degenerated nucleus pulposus tissue and NPCs;Inhibition of Gli1 gene expression downregulates the expressions of FAP and FSP-1 proteins.The mechanism of intervertebral disc fibrosis and degeneration may be associated with the activation of Gli1.

Objectives:To explore the impact of Gli1(gioma-associated oncogene homolog)on nucleus pulposus cells(NPCs)and its role in the fibrotic changes of the nucleus within intervertebral discs.Methods:A rat tail intervertebral disc puncture model was established.Histological staining was used to evaluate the levels of intervertebral disc degeneration and fibrotic changes in the untreated(negative control,NC)group and the puncture(puncture-induced intervertebral disc degeneration,PIDD)group.Western blot was used to assess the expression levels of Gli1 protein in the tissues.Gli1 protein was engineered to overexpress by lentivirus in NPCs and verified.The experiment was divided into three groups:control(Blank),Gli1 overexpression(Lv-Gli1),and virus control(lv-Ctrl).Immunofluorescence and Western blot were used to detect the expression levels of fibrotic markers FAP and FSP-1.Adding Gli1 inhibitor GANT61,cellular experiment was grouped into control(blank),Gli1 overexpression(Lv-Gli1),Gli1 overexpression+GANT61(Lv-Gli1+GANT61),and Gli1 overexpression+DMSO(Lv-Gli1+DMSO).Western blot was used to detect the expression levels of FAP and FSP-1 proteins.In vivo animal experiments were conducted in groups of puncture(PIDD),puncture treatment(PIDD+GANT61),and treatment control(PIDD+DMSO).Histological staining was used to evaluate the levels of intervertebral disc degeneration and fibrotic changes.Results:Histological staining results showed the histological score of degenerated discs significantly increased(P＜0.01).Western blot results showed a significant increase in Gli1 protein expression in the degenerated group(P＜0.05),which was consistent with the conclusions of the cellular experiments.NPCs overexpressing Gli1 protein were constructed,and immunofluorescence staining results showed that the expression of FAP and FSP-1 in the Gli1 overexpression group was significantly upregulated compared to the control and virus control groups.Western blot results further showed a significant increase in the expression of fibrotic markers in the Gli1 overexpression group(P＜0.05).In the Gli1 overexpression+GANT61 group of the cellular experiment,compared to the control and Gli1 overexpression groups,Western blot indicated that the expressions of FAP and FSP-1 proteins were downregulated(P＜0.05).In the in vivo experiment,the PIDD+GANT61 group showed a significant improvement in histological scores and collagen composition(P＜0.05).Conclusions:The expression of Gli1 protein is significantly upregulated in degenerated nucleus pulposus tissue and NPCs;Inhibition of Gli1 gene expression downregulates the expressions of FAP and FSP-1 proteins.The mechanism of intervertebral disc fibrosis and degeneration may be associated with the activation of Gli1.

Objective To analyze the factors associated with pain after arthroscopic rotator cuff bridge suture.Methods According to the inclusion and exclusion criteria,the data of 112 patients with unilateral rotator cuff injury who received arthroscopic bridge suture in our department were collected and were investigated in the form of telephone follow-up.In this study,SPSS 23.0 was used to input data and conduct statistical analysis.Logistic regression analysis was used to analyze the correlation between the above influencing factors and postoperative pain.Results A total of 112 patients were included for statistical analysis,single factor analysis revealed,including course of disease,smoking history,preoperative University of California,Los Angeles(UCLA)score,Constant score,numeric rating scale(NRS),size of rotator cuff tear,whether it was full-thickness tear and degree of tendon retraction might be related to postoperative pain(P<0.05).The age,gender,body mass index(BMI),drinking history,diabetes and hypertension were not related to postoperative pain(P>0.05).Multiple linear regression analysis concluded that there were four factors related to postoperative pain,and the correlation degree was preoperative NRS,preoperative UCLA score,tear size and smoking history.Conclusion The causes of postoperative pain after arthroscopic rotator cauff repair are complex and diverse.Analyzing the cause of postoperative pain can effectively reduce the pain of patients and promote the recovery of shoulder joint function.

Objectives:To compare the short-term effectiveness and safety between robot-assisted cortical bone trajectory screw(CBTS)and traditional pedicle screw(TPS)fixations in posterior lumbar interbody fusion(PLIF)for lumbar spinal stenosis with osteoporosis.Methods:We retrospectively analyzed the clinical data of 113 patients who underwent robot-assisted PLIF and internal fixation in our hospital between September,2018 and June,2021.The patients were divided into two groups according to the internal fixation method.55 pa-tients fixed with CBTS were included in the CBTS group,and 58 patients fixed with TPS were included in the TPS group.The general data[age,sex ratio,height,weight,body mass index(BMI),hip bone mineral densi-ty(BMD)],surgical related data(operative time,incision length,intraoperative blood loss),perioperative indicators(postoperative length of hospital stay,surgical complications,screw loosening rate,adjacent facet joint violation rate,preoperative and postoperative blood glucose),and visual analogue scale(VAS)score of pain and Oswestry disability index(ODI)before and after surgery were recorded and compared.Results:There was no significant difference in age,sex ratio,height,weight,BMI and hip BMD between the two groups(P＞0.05).There was no significant difference in operative time,postoperative length of hospital stay,surgical complications,preoperative VAS score and ODI,and ODI at 1 month and 6 months after operation between the two groups(P＞0.05).The intraoperative blood loss,length of incision and VAS score at 6 months after operation in the CBTS group were significantly lower than those in the TPS group(500mL vs 600mL,P=0.007;8cm vs 10cm,P=0.006;2 vs 3,P＜0.001).There was no significant difference in blood glucose before operation and 1 day after operation between the two groups(P＞0.05),while at 1 week after operation,the CBTS group was significantly lower than the TPS group(4.75mmol/L vs 5.57mmol/L,P=0.041).At the final follow-up,the rates of screw loosening and adjacent facet joint violation in CBTS group were significantly lower than those in TPS group(1.68％vs 4.52％,P=0.044 and 13.83％vs 26.53％,P=0.029).Conclusions:For patients with lumbar spinal stenosis combined with osteoporosis,PLIF with robot-assisted CBTS fixation is less in surgical trauma,faster in early postoperative recovery,and fewer in screw-related complications than with TPS fixation.

Objective To investigate the protective effect of silymarin extract(SME)and its complex preparation on ethanol liver injury.Methods An ethanol liver injury model was established by gavage of 12 mL·kg-1 50％ethanol.Male mice were divided into blank group(distilled water),model group(ethanol liver injury model),SME-L,-H groups(6,20 mg·mL-1 SME),SME+Ganoderma lucidum extract(GLE)-L,-H groups(10,30 mg·mL-1 SME+GLE,SME∶GLE=1∶1),Jian An Shi Silymarin Pueraria Mirifica and Tansy tablets(JAS)-L,-H groups(68,204 mg·mL-1 JAS),there were 12 mice in each group.The serum levels of glutamic-oxaloacetic transaminase(GOT)in mice were measured by fully automated biochemical analyzer assay;the serum levels of interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)in mice were measured by enzyme-linked immunosorbent assay(ELASA);the hepatic tissue of oxidative stress indexes[catalase(CAT)and total superoxide dismutase(T-SOD)]were measured by ultraviolet spectrophotometer.Results The T-SOD activity in the blank group,model group,SME-L,SME-H,SME+GLE-L,SME+GLE-H,JAS-L and JAS-H groups were(192.54±49.00),(141.65±34.72),(205.83±32.77),(191.68±25.83),(192.31±28.79),(177.82±32.61),(218.58±74.80)and(210.24±31.65)U·mg·prot-1;CAT activity were(37.78±5.73),(28.92±8.44),(44.12±11.52),(41.41±9.15),(47.01±10.48),(41.63±8.95),(47.14±8.91)and(48.29±10.06)U·mg-1;GPT levels were(47.61±13.00),(97.84±26.00),(62.33±18.92),(51.84±17.91),(70.77±28.00),(58.00±21.27),(52.28±18.78)and(45.55±9.27)U·L-1;IL-6 levels were(21.03±1.52),(28.43±5.75),(21.90±3.24),(21.23±1.55),(22.26±2.58),(21.24±2.91),(22.17±4.14)and(21.14±3.02)pg·mL-1.Comparing the above indexes in the model group with the blank group,and comparing the above indexes in the SME-L,SME-H,SME+GLE-L,SME+GLE-H,JAS-L,JAS-H groups with the model group,the differences were statistically significant(all P＜0.01).The TNF-α levels in blank,model,SME-L,SME-H,SME+GLE-L,SME+GLE-H,JAS-L and JAS-H groups were were(28.07±7.72),(69.02±16.34),(40.29±8.94),(48.84±10.17),(41.91±14.96),(40.07±12.75),(50.72±11.44)and(45.05±11.34)pg·mL-1.Comparing the model group with the blank group,the SME,SME+GLE-L,-M and JAS,-M groups with the model group,the differences were statistically significant(all P＜0.01).Conclusion Silybum marianum extract and its compound preparation can increase the antioxidant level and reduce the inflammation of mouse liver,and have a certain improvement effect on liver injury caused by acute ethanol poisoning.

Objectives:To compare the short-term effectiveness and safety between robot-assisted cortical bone trajectory screw(CBTS)and traditional pedicle screw(TPS)fixations in posterior lumbar interbody fusion(PLIF)for lumbar spinal stenosis with osteoporosis.Methods:We retrospectively analyzed the clinical data of 113 patients who underwent robot-assisted PLIF and internal fixation in our hospital between September,2018 and June,2021.The patients were divided into two groups according to the internal fixation method.55 pa-tients fixed with CBTS were included in the CBTS group,and 58 patients fixed with TPS were included in the TPS group.The general data[age,sex ratio,height,weight,body mass index(BMI),hip bone mineral densi-ty(BMD)],surgical related data(operative time,incision length,intraoperative blood loss),perioperative indicators(postoperative length of hospital stay,surgical complications,screw loosening rate,adjacent facet joint violation rate,preoperative and postoperative blood glucose),and visual analogue scale(VAS)score of pain and Oswestry disability index(ODI)before and after surgery were recorded and compared.Results:There was no significant difference in age,sex ratio,height,weight,BMI and hip BMD between the two groups(P＞0.05).There was no significant difference in operative time,postoperative length of hospital stay,surgical complications,preoperative VAS score and ODI,and ODI at 1 month and 6 months after operation between the two groups(P＞0.05).The intraoperative blood loss,length of incision and VAS score at 6 months after operation in the CBTS group were significantly lower than those in the TPS group(500mL vs 600mL,P=0.007;8cm vs 10cm,P=0.006;2 vs 3,P＜0.001).There was no significant difference in blood glucose before operation and 1 day after operation between the two groups(P＞0.05),while at 1 week after operation,the CBTS group was significantly lower than the TPS group(4.75mmol/L vs 5.57mmol/L,P=0.041).At the final follow-up,the rates of screw loosening and adjacent facet joint violation in CBTS group were significantly lower than those in TPS group(1.68％vs 4.52％,P=0.044 and 13.83％vs 26.53％,P=0.029).Conclusions:For patients with lumbar spinal stenosis combined with osteoporosis,PLIF with robot-assisted CBTS fixation is less in surgical trauma,faster in early postoperative recovery,and fewer in screw-related complications than with TPS fixation.

To revise GBZ 188 Technical Specification for Occupational Health Surveillance based on national laws, regulations, standards, specifications and legal documents of occupational disease, and combination with the actual situation in China. The main modifications are as follows: the occupational health surveillance for workers exposed to toluene (xylene may implement by reference), bromopropane, methyl iodide, ethylene oxide, chloroacetic acid, indium and its compounds, coal tar, coal tarasphalt, asphalt, β-naphthylamine, dust of metal and its compounds(tin, iron, antimony, barium and its compounds), hard metal dust, erionite dust, low temperature, laser, tick-borne encephalitis virus, Borrelia burgdorferi, and human immunodeficiency virus, for scraper or grind operators, and underground workers using squatting or kneeling position, crawling position, side-lying position, or shoulder position for a long period of time are included. The emergency health screening for workers exposed to arsenic, fluorine and its inorganic compounds, and acrylamide are included. The occupational medical examination (OME) for workers exposed to amino and nitro compounds of benzene, phosgene, monomethylamine, organic fluorine and dimethyl sulfate has been adjusted and made mandatory, with corresponding assessments required upon leaving the job. The special occupational health surveillance for workers exposed to mycobacterium tuberculosis and hepatitis virus is removed. The OME conclusion of reexamination is removed, and standardize recheck/additional inspection requirements. The optional items in OME performed before, during and after leaving post are removed, but the optional items in emergency medical examination are retained. Additional OME items are added. The Guideline for OME Summary Reports is added as informative appendix, and so on. The revised GBZ 188 Technical Specification for Occupational Health Surveillance is more scientific and practical.

Objective To investigate the effect and underlying mechanism of SULT2B1 in the de-velopment of atherosclerosis(AS).Methods Twelve 8-week-old apolipoprotein E-knockout(apoE-/-)male mice were subjected and fed with a high-fat diet for 12 weeks,and then randomly divided into adeno-associated virus(AAV)-GFP and AAV-shSULT2B1 groups,with 6 animals in each group.In 4 weeks after AAV injection via tail vein,the mice were sacrificed for assessing aortic and aortic root plaque formation by oil red O staining and detecting serum levels of inflam-matory factors and blood lipids.RAW264.7 cells were transfected with adenovirus(Ad)-GFP and Ad-SULT2B1,respectively(n=3).RNA sequencing was performed to detect downstream RNA changes.Then LncRNA gga3-204 was selected for downstream study.After RAW264.7 cells were divided into si-NC group,si-SULT2B1 group,si-Lncgga3-204 group and si-SULT2B1+si-Lncg-ga3-204 group(n=3),and the IL-1β and IL-6 levels were detected in these transfected cells.Results There was no statistically difference in body weight in the mice from the AAV-GFP and AAV-shSULT2B1 groups after high-fat feeding(P＞0.05).Significantly lower serum levels of TC,TG and LDL-C,reduced aortic plaque area[(8.38±1.33)％vs(11.83±1.04)％,P=0.000],and decreased TG content within the aortic root plaque[(12.29±1.54)％vs(17.67±1.53)％,P=0.000]were observed in the AAV-shSULT2B1 group than those in the AAV-GFP group.Ser-um IL-1β and IL-6 levels in the mice of the AAV-shSULT2B1 group than those in the AAV-GFP group(P＜0.01).The AAV-shSULT2B1 group also had obviously lower serum levels of I L-1 βand IL-6 than the AAV-GFP group(P＜0.01).In the RAW264.7 cells from the si-SULT2B1 group,the mRNA levels of IL-1β and IL-6 were notably lower than those in the si-NC group(P＜0.01).LncRNA gga3-204 expression was significantly higher in the Ad-shSULT2B1 group than the Ad-GFP group(P＜0.01).While,the si-SULT2B1 group had statistically higher Lncgga3-204 level than the si-NC group(2.32±0.60 vs 1.19±0.21,P=0.036).The si-Lncgga3-204 group had significantly higher IL-1β and IL-6 mRNA levels than the si-NC group(P＜0.01).The si-SULT2B1+si-Lncgga3-204 group had significantly higher IL-1β and IL-6 mRNA levels than the si-SULT2B1 group(P＜0.05).Conclusion SULT2B1 affects the macrophage inflammatory response via Lncgga3-204,and then affects the progression of AS.

Biological matrix reference material is a reference material that combines the target material with the biological matrix. The biological matrix reference material has higher consistency with the authentic specimens in forensic toxicology, and its application has a positive effect on improving the accuracy of test results. This paper reviews the research on the matrix reference materials corresponding to three common biological test materials (blood, urine and hair). In order to provide reference for the development and application of biological matrix reference materials in forensic toxicology, this paper mainly introduces the research progress of preparation technology of biological matrix reference materials and some existing products and their parameters evaluation.
Forensic Toxicology/methods* ; Hair ; Body Fluids

This paper reviews the status quo of recent years’ research on autophagy and damage of chondrocytes in Kashin-Beck disease （KBD） and osteoarthritis （OA）. PI3K/AKT signaling pathway and mTOR regulate the autophagy activity of chondrocytes. PI3K/AKT signaling pathway can promote the proliferation of chondrocytes and cause their damage by inhibiting their autophagy activity. This might play an important role in the occurrence and progression of bone and joint diseases such as KBD and OA， and provide scientific basis for revealing the pathogenesis and treatment plan of them.