OBJECTIVE To establish skin contamination models with sulfur mustard analogue 1-chloro-2-ethylsulfanyl ethane(CEES),and evaluate the therapeutic efficacy of reactive skin decontamination lotion kit(RSDL).METHODS ①Kunming,BALB/c,BALB/c-nu,and C57BL/6N were contaminted with CEES 75 μL·kg-1 using exposed method and covered method for 10 min on the dorsal skin.Wound healing times and areas were assessed to determine the stock of mice and exposure method.② BALB/c mice were exposed to a gradient of CEES at the doses of 75,150,250,350 and 500 μL·kg-1(10 min)using the covered method.Wound healing times,areas and Kaplan-Meier survival curves were used to determine the optimal dose.③ BALB/c mice were exposed to CEES 150 μL·kg-1 for varying durations(5,10 and 20 min)using the covered method.Wound healing times,areas and Kaplan-Meier survival curves were analyzed to determine the optimal exposure time.④ BALB/c mice were divided into four groups:control(exposed with the covered method at 150 μL·kg-1,10 min),treatment(20 μL RSDL treatment after exposure,as the control group),5 min treatment(20 μL RSDL treatment after 5 min of exposure,followed by 5 min of coverage)and immediate-treatment(exposed with the exposed method at 150 μL·kg-1,immediately treated with 20 μL RSDL,followed by 10 min of coverage).The therapeutic efficacy was evaluated based on the wound area,subcutaneous microvesicle count,epidermal thick-ness,and inflammatory cytokine(IL-6 and TNF-α)expressions.RESULTS ① The covered method caused more severe and prolonged wounds than the exposed method.BALB/c mice exhibited a high sensitivity to CEES with delayed wound healing and were therefore selected as the model animal.② Survival rates in BALB/c mice dropped below 50%at doses of 250,350 and 500 μL·kg-1,whereas an 83.3%survival rate was observed at 150 μL·kg-1.③ The mice exposed to CEES(150 μL·kg-1,20 min)died within 3 days.The wound area was consistently smaller in the 5 min covered group than in the 10 min covered group.④CEES-induced skin injury led to epidermal nuclear pyknosis,follicular disrup-tion,inflammatory infiltration,and microvesicle formation in both treatment and poisoned control groups.As more immediate treatment,the wound area significantly decreased.While the IL-6 expres-sion showed no significant intergroup difference,the TNF-α expression was significantly higher in the treatment group.CONCLUSION A CEES-induced skin contamination model has been established in BALB/c mice using the covered method(150 μL·kg-1,10 min covered).However,RSDL should be ad-ministered in 10 min post-contamination.

Dermacentor abaensis(D.abaensis)is a tick species distributed only in Qinghai,Gansu,and other provinces of China.It is more harmful because it can carry pathogens such as Borrelia burgdorferi and Piroplasm.However,morphological descriptions and image data for this tick spe-cies are still insufficiently detailed,and data related to molecular markers such as COX 1 and ITS2 are still missing.The ticks were collected from the body surface of yaks in Diebu County,Gannan Tibetan Autonomous Prefecture,Gansu Province.The morphological characteristics of various parts of the ticks were carefully observed using a stereomicroscope ultra-depth-of-field system.At the same time,the total DNA of the tick body wall was extracted,then the primers for COX 1 and ITS2 genes were designed,and PCR amplification was performed before sequencing.The sequen-cing results were compared with related sequences in GenBank for homology analysis,and the phy-logenetic tree was constructed to provide a basis for accurate identification of this tick species.The results showed that the female ticks were subrounded.The basic capituli were rectangular in shape.The porose area was ovoid in shape,cornua short and blunt,palps thick and short,with a slightly pointed front end.A slightly rounded scutum,and the enamel color was slightly lighter towards the back.The cervial groove was short,and in deeply concave.The festoon was obvious.The peritreme was in comma-shaped.The trailing edge was slightly curved.The genital opening had a ala.The anus was located in the posterior quarter of the middle of the abdomen,and there is a semicircular anal groove surrounded.Male ticks were slightly elongated ovoid,with no porose area.The entire surface was covered with enamel color except the cornua.The genital opening was opposite to the coxall,and there is no ala.The anal groove was deeper,and the external spur of the coxa Ⅳ was narrower and longer.All other body characteristics are similar to those of female ticks.Based on the comparative analysis of COX 1 and ITS2 gene sequences and the construction of phylogenetic tree,it was found that D.abaensis is more closely related to Dermacentor nuttalli and Dermacentor marginatus than other Dermacentor spp.The results suggested that D.abaensis have unique mor-phological and molecular marker characteristics,and the combination of the two characteristics will make it easier to rapidly and accurately identify this tick species.

Dermacentor abaensis(D.abaensis)is a tick species distributed only in Qinghai,Gansu,and other provinces of China.It is more harmful because it can carry pathogens such as Borrelia burgdorferi and Piroplasm.However,morphological descriptions and image data for this tick spe-cies are still insufficiently detailed,and data related to molecular markers such as COX 1 and ITS2 are still missing.The ticks were collected from the body surface of yaks in Diebu County,Gannan Tibetan Autonomous Prefecture,Gansu Province.The morphological characteristics of various parts of the ticks were carefully observed using a stereomicroscope ultra-depth-of-field system.At the same time,the total DNA of the tick body wall was extracted,then the primers for COX 1 and ITS2 genes were designed,and PCR amplification was performed before sequencing.The sequen-cing results were compared with related sequences in GenBank for homology analysis,and the phy-logenetic tree was constructed to provide a basis for accurate identification of this tick species.The results showed that the female ticks were subrounded.The basic capituli were rectangular in shape.The porose area was ovoid in shape,cornua short and blunt,palps thick and short,with a slightly pointed front end.A slightly rounded scutum,and the enamel color was slightly lighter towards the back.The cervial groove was short,and in deeply concave.The festoon was obvious.The peritreme was in comma-shaped.The trailing edge was slightly curved.The genital opening had a ala.The anus was located in the posterior quarter of the middle of the abdomen,and there is a semicircular anal groove surrounded.Male ticks were slightly elongated ovoid,with no porose area.The entire surface was covered with enamel color except the cornua.The genital opening was opposite to the coxall,and there is no ala.The anal groove was deeper,and the external spur of the coxa Ⅳ was narrower and longer.All other body characteristics are similar to those of female ticks.Based on the comparative analysis of COX 1 and ITS2 gene sequences and the construction of phylogenetic tree,it was found that D.abaensis is more closely related to Dermacentor nuttalli and Dermacentor marginatus than other Dermacentor spp.The results suggested that D.abaensis have unique mor-phological and molecular marker characteristics,and the combination of the two characteristics will make it easier to rapidly and accurately identify this tick species.

OBJECTIVE To establish skin contamination models with sulfur mustard analogue 1-chloro-2-ethylsulfanyl ethane(CEES),and evaluate the therapeutic efficacy of reactive skin decontamination lotion kit(RSDL).METHODS ①Kunming,BALB/c,BALB/c-nu,and C57BL/6N were contaminted with CEES 75 μL·kg-1 using exposed method and covered method for 10 min on the dorsal skin.Wound healing times and areas were assessed to determine the stock of mice and exposure method.② BALB/c mice were exposed to a gradient of CEES at the doses of 75,150,250,350 and 500 μL·kg-1(10 min)using the covered method.Wound healing times,areas and Kaplan-Meier survival curves were used to determine the optimal dose.③ BALB/c mice were exposed to CEES 150 μL·kg-1 for varying durations(5,10 and 20 min)using the covered method.Wound healing times,areas and Kaplan-Meier survival curves were analyzed to determine the optimal exposure time.④ BALB/c mice were divided into four groups:control(exposed with the covered method at 150 μL·kg-1,10 min),treatment(20 μL RSDL treatment after exposure,as the control group),5 min treatment(20 μL RSDL treatment after 5 min of exposure,followed by 5 min of coverage)and immediate-treatment(exposed with the exposed method at 150 μL·kg-1,immediately treated with 20 μL RSDL,followed by 10 min of coverage).The therapeutic efficacy was evaluated based on the wound area,subcutaneous microvesicle count,epidermal thick-ness,and inflammatory cytokine(IL-6 and TNF-α)expressions.RESULTS ① The covered method caused more severe and prolonged wounds than the exposed method.BALB/c mice exhibited a high sensitivity to CEES with delayed wound healing and were therefore selected as the model animal.② Survival rates in BALB/c mice dropped below 50%at doses of 250,350 and 500 μL·kg-1,whereas an 83.3%survival rate was observed at 150 μL·kg-1.③ The mice exposed to CEES(150 μL·kg-1,20 min)died within 3 days.The wound area was consistently smaller in the 5 min covered group than in the 10 min covered group.④CEES-induced skin injury led to epidermal nuclear pyknosis,follicular disrup-tion,inflammatory infiltration,and microvesicle formation in both treatment and poisoned control groups.As more immediate treatment,the wound area significantly decreased.While the IL-6 expres-sion showed no significant intergroup difference,the TNF-α expression was significantly higher in the treatment group.CONCLUSION A CEES-induced skin contamination model has been established in BALB/c mice using the covered method(150 μL·kg-1,10 min covered).However,RSDL should be ad-ministered in 10 min post-contamination.

Human beings are inevitably exposed to toxic substances as a result of influences of potential contamination factors in the environment,food and medicines,which poses a threat to human health.In order to effectively screen and prevent the exposure or intake of such substances,it is neces-sary to develop in vitro assays for the detection and toxicity evaluation of toxic substances.High content screening(HCS)has been recognized as an important tool for toxicity testing and risk assessment of compounds due to its high throughput and automation advantages,and has been widely used in in vitro toxicology research.In this review,we described the system components of HCS and its workflow in toxicity screening and toxicity evaluation by focusing on cases of their application in toxicity detection and evaluation studies,including the cytotoxicity,hepatotoxicity,nephrotoxicity,genotoxicity,neurotox-icity,cardiotoxicity,and developmental toxicity.In addition,the applications and developments of machine learning in HCS were explored,especially to the advantages of supervised and unsupervised machine learning strategies for high throughput image screening and data analysis.Finally,the future applications of HCS in toxicity screening and evaluation are outlined,especially in terms of binding new models and gene editing technology.

Objective:To analyze the ideal entry parameters and entry points for C 7 pedicle screws based on three-dimensional CT reconstruction model. Methods:A retrospective case series study was used to analyze the cervical spine CT image data of 50 adult volunteers collected from April 2021 to March 2022 at Taizhou People′s Hospital Affiliated to Nanjing Medical University, including 25 males and 25 females; aged 20-67 years [(43.8±13.5)years]. No significant structural abnormalities were seen in the cervicothorax of all volunteers. First, the 50 CT images were imported into the medical engineering software Mimics 17.0 to reconstruct the three-dimensional C 7 model, and the C 7 pedicle screw diameter (PSD), pedicle screw length (PSL), sagittal angle (SA) and lateral angle (LA) were measured. Then, the anatomical markers, namely the lateral notch and the midpoint of the inferior edge of the C 6 articular process, were used to measure the horizontal distance between the midpoint of the inferior edge of the C 6 articular process and the ideal entry point (line segment A), and the horizontal and vertical distances between the lateral notch and the ideal entry point (line segments B and C). Finally, the lateral notch and the midpoint of the inferior edge of the C 6 articular process were used as markers to observe the distribution of entry points. The values of the above measured parameters were recorded and compared with each other to analyze the differences between different sides and genders. Intra-group correlation coefficients (ICC) were also used to assess intra- and inter-observer agreement. Results:All 100 pedicles from 50 C 7 models were accessed, with the PSD being (6.5±0.7)mm, PSL being (31.8±4.5)mm, SA being (89.8±8.8)°, LA being (31.0±6.7)°, line segment A being 0.9(-0.4, 2.1) mm, line segment B being (5.8±1.7)mm, and line segment C being (3.6±1.5)mm. All ideal entry points were located medial above the lateral notch; moreover, with the midpoint of the inferior edge of the C 6 articular process as a reference, 71 (71%) of the entry points were located laterally and 29 (29%) were located medially. The 12 mm area around the midpoint of the inferior edge of the C 6 articular process was divided into 6 sections, with 47% of the entry points being located within 2 mm lateral to the midpoint, 25% within 2 mm medial to the midpoint, 19% within 2-4 mm lateral to the midpoint, and only 9% within 50% lateral to the midpoint. The measured parameters were not significantly different between the left and right sides (all P>0.05). The PSD, SA, and line segment B in males were (6.8±0.7)mm, (92.0±8.2)°, and (6.3±1.6)mm, which were significantly greater than those in females [(6.2±0.6)mm, (87.5±8.0)°, and (5.3±1.6)mm] (all P<0.01). The remaining parameters were similar between two genders (all P>0.05). All measured parameters had high intra- and inter-observer agreement (ICC: 0.84-0.91), except for line segment A that had moderate intra- and inter-observer agreement (ICC: 0.46-0.63). Conclusions:For C 7 pedicle screw implantation, when the surface of the lateral mass is used as the reference plane, SA is approximately 90°; with reference to the lateral notch, all the ideal entry points are located medially above it; with reference to the midpoint of the inferior edge of the C 6 articular process, approximately 70% of the entry points are located laterally. There are no significant differences in the parameters between the different sides of screw implantation on pedicles. The measured parameters show high intra- and inter-observer agreement, except for line segment A.

Objective: This study’s objective is to assess the efficacy and safety of Pulsed Magnetic Therapy System (PMTS) in improving insomnia disorder. Methods: Participants with insomnia disorder were randomly assigned to receive either PMTS or sham treatment for four weeks (n= 153; PMTS: 76, sham: 77). Primary outcomes are the Insomnia Severity Index (ISI) scores at week 0 (baseline), 1, 2, 3, 4 (treatment), and 5 (follow-up). Secondary outcomes are the Pittsburgh Sleep Quality Index at baseline and week 4, and weekly sleep diary-derived values for sleep latency, sleep efficiency, real sleep time, waking after sleep onset, and sleep duration. Results: The ISI scores of the PMTS group and the sham group were 7.13±0.50, 11.07±0.51 at week 4, respectively. There was a significant group×time interaction for ISI (F3.214, 485.271=24.25, p<0.001, ηp 2=0.138). Only the PMTS group experienced continuous improvement throughout the study; in contrast, the sham group only experienced a modest improvement after the first week of therapy. At the end of the treatment and one week after it, the response of the PMTS group were 69.7% (95% confidence interval [CI]: 58.6%–79.0%), 75.0% (95% CI: 64.1%–83.4%), respectively, which were higher than the response of the sham group (p<0.001). For each of the secondary outcomes, similar group×time interactions were discovered. The effects of the treatment persisted for at least a week. Conclusion PMTS is safe and effective in improving insomnia disorders.

Objective:To explore the effect of hydrogen sulfide (H 2S) on expression of phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) signal pathway in rats with intestinal ischemia/reperfusion (IRI) injury. Methods:Thirty male Wistar rats were divided into sham operation group (Sham group), IRI group, and H 2S precursor sodium hydrosuphide (NaHS) intervention group (IRI+NaHS group) by random number table method, with 10 rats in each group. The animal model of IRI was established by 60 minutes superior mesenteric artery (SMA) blockage with non-invasive vascular clamp and 120 minutes reflow. SMA was dissociated and peritoneum cavity was closed in Sham group. The rats in IRI+NaHS group was received NaHS (100 μmol/kg bolus+1.07 mmol·kg -1·h -1 infusion) 10 minutes prior to the onset of reperfusion, while the rats in IRI group and Sham group were received equal volume of normal sodium. Blood in vena cava was collected. H 2S was detected by sensitive sulfide electrode. Rats were sacrificed after blood collection. Histopathology change was observed by hematoxylin-eosin (HE) staining, ileal pathological score was studied by Chiu score. The protein expressions of phosphated Akt (p-Akt), Akt, PI3K, cleaved caspase-9, mammalian target of rapamycin (mTOR) were determined by Western Blot. Results:Compared with the Sham group, there was intestinal mucosa structure disorder edema and shedding villous fracture in the IRI group. Ileal pathological score in IRI group was significantly increased (4.21±0.15 vs. 0.15±0.03, P < 0.01), while plasma H 2S in IRI group was significantly decreased (μmol/L: 26.72±3.17 vs. 38.34±5.24, P < 0.01). Ileal p-Akt, PI3K, caspase-9 and mTOR protein in IRI group were significantly increased (p-Akt/GAPDH: 2.67±0.12 vs. 0.24±0.05, PI3K/GAPDH: 1.42±0.07 vs. 0.57±0.08, caspase-9/GAPDH: 4.23±0.61 vs. 0.13±0.02, mTOR/GAPDH: 2.17±0.23 vs. 0.23±0.02, all P < 0.01). Compared with the IRI group, pathological changes of intestinal mucosa in the IRI+NaHS group was improved, ileal pathological score was significantly decreased (1.56±0.02 vs. 4.21±0.15, P < 0.01), plasma H 2S was significantly increased (μmol/L: 32.36±2.45 vs. 26.72±3.17, P < 0.01) and ileal p-Akt, PI3K were significantly increased (p-Akt/GAPDH: 5.12±0.08 vs. 2.67±0.12, PI3K/GAPDH: 3.14±0.05 vs. 1.42±0.07, both P < 0.01), while caspase-9, mTOR in IRI+NaHS group were significantly decreased (caspase-9/GAPDH: 2.12±0.24 vs. 4.23±0.61, mTOR/GAPDH: 1.37±0.28 vs. 2.17±0.23, both P < 0.01). Conclusion:H 2S attenuates intestinal injury in IRI rats by up-regulating PI3K/Akt signal pathway and down-regulating caspase-9 and mTOR.

Objective:To investigate the effects of acupoint massage combined with high-frequency repetitive transcranial magnetic stimulation on post-stroke depression and quality of life.Methods:A total of 169 patients with post-stroke depression who were admitted to the Department of Neurological Intensive Care Unit of Zhengzhou University First Affiliated Hospital from June 2017 to June 2019 were selected as the research subjects. The patients were divided into observation group (85 cases) and control group (84 cases) by random number table method. The control group was treated with high frequency repetitive transcranial magnetic stimulation, and the observation group was treated with high frequency repetitive transcranial magnetic stimulation combined with acupoint massage, high frequency repetitive transcranial magnetic stimulation 5 times a week, 28 minutes each time, 8 weeks of intervention, acupoint massage once a day, 1-2 minutes each acupoint, 8 weeks of intervention. The state of depression and the improvement of quality of life were observed, and the adverse reactions were recorded.Results:The total effective rate of depressive symptoms improvement in the observation group was 83.53% (71/85), which was higher than that of the control group 70.24% (59/84), the difference was statistically significant ( χ 2 value was 4.204, P<0.05). After the intervention, the Hamilton Depression (HAMD) scores of both groups decreased, and the trend decreased with time, and the decline of the observation group was higher than that of the control group, the difference was statistically significant ( Ftime value was 888.220, Fgroup value was 191.331, Finteraction value was 14.955, P<0.01 or 0.05). The observation group ′s physiological function, physiological function, energy, social function, general health status, emotional function, and mental health score were (57.25 ± 10.33), (64.41 ± 13.35), (67.26 ± 12.93), (70.43 ± 14.15), (64.64 ± 12.52), (72.71 ± 13.24), (75.24 ± 12.90) points, higher than the control group ′s (51.83 ± 10.22), (55.40 ± 12.90), (62.84 ± 12.81), (66.03 ± 13.94), (59.37 ± 11.26), (67.54 ± 13.07), (70.85 ± 12.66) points, the difference was statistically significant ( t value was 2.036-4.461, P<0.01 or 0.05). There was no significant difference in the incidence of adverse reactions between the two groups ( P > 0.05). Conclusions:Acupoint massage combined with high-frequency repetitive transcranial magnetic stimulation has a effective in the treatment of post-stroke depression, which can significantly improve the depression status and quality of life.

Objective: To investigate the effect of three-dimensional conformal radiotherapy (3D-CRT) combined with PC chemotherapy (paclitaxel + carboplatin) on non-small cell lung cancer (NSCLC) patients and the serum levels of CA125, tissue inhibitor of metalloproteinase-1 (TIMP-1), serum amyloid A (SAA) and T-lymphocyte subsets. Methods: A total of 100 patients with NSCLC treated in Affiliated Hospital of Guangdong Medical University from May 2015 to December 2017 were selected as the study subjects. They were divided into control group and observation group according to random number table method, with 50 cases in each group. The observation group was treated with 3D-CRT combined with PC chemotherapy, while the control group was treated with PC chemotherapy. The two groups were treated for 4 cycles. The therapeutic effect, serum CA125, TIMP-1, SAA, T-lymphocyte subsets and adverse reactions were compared between the two groups. Results: Four cases were lost to follow-up both in the two groups. The overall response rate in the observation group (43.48%, 20/46) was higher than that in the control group (23.91%, 11/46; χ²=3.941, P=0.047). The serum levels of CA125, TIMP-1 and SAA of the two groups had no significant difference before treatment, and the levels of these indexes decreased after treatment. The serum levels of CA125, TIMP-1 and SAA in the observation group after treatment were (12.31±1.13) U/ml, (275.31±13.69) pg/ml and (47.21±7.21) mg/L, which were lower than those in the control group [(30.36±1.98) U/ml, (320.36±17.23) pg/ml, (65.92±8.36) mg/L], with significant differences (t=53.699, P<0.001; t=13.884, P<0.001; t=11.495, P<0.001). The levels of CD3⁺ , CD4⁺ , CD8⁺ and CD4⁺ /CD8⁺ of the two groups had no significant difference before treatment, and the levels of these indexes decreased after treatment. The levels of CD3⁺ , CD4⁺ , CD8⁺ and CD4⁺ /CD8⁺ in the observation group were (35.27±10.31)%, (20.27±6.72)%, (15.89±3.37)% and 0.91±0.37, which were higher than those in the control group [(30.77±9.27)%, (15.27±5.73)%, (12.02±2.69)% and 0.75±0.39], with significant differences (t=2.201, P=0.030; t=3.840, P<0.001; t=6.087, P<0.001; t=2.019, P=0.047). There were no significant differences in the adverse reactions such as nausea and vomiting [63.04% (29/46) vs. 43.48% (20/46); χ²=3.537, P=0.060], phlebitis [6.52% (3/46) vs. 4.35% (2/46); χ²=0.000, P>0.999], abnormal liver function [6.52% (3/46) vs. 2.17% (1/46); χ²=0.261, P=0.609] and myelosuppression [8.70% (4/46) vs. 6.52% (3/46); χ²=0.000, P>0.999] between the observation group and the control group. Conclusion For patients with NSCLC, 3D-CRT combined with PC chemotherapy can improve the overall response rate, decrease the levels of serum CA125, TIMP-1 and SAA, and improve the immune function of patients. The therapeutic effect is remarkable and the safety is good. The therapeutic scheme is suitable for the treatment of NSCLC.