Objective:To investigate the effect of PD-1/PD-L1 inhibitor BMS-1 on LPS-induced inflammation in mouse microg-lia cells(BV-2 cells).Methods:Bv-2 cells were divided into Control group,LPS group and LPS+BMS-1 group.Bv-2 cells in Control group were cultured in DMEM medium for 78 hours,cells in LPS group were stimulated with 100 ng/ml LPS for 6 hours after 72 hours of normal culture,Bv-2 cells in LPS+BMS-1 group were treated with 50 nmol/ml BMS-1 for 72 hours and then stimulated with 100 ng/ml LPS for 6 hours.Expressions of PD-1 and iNOS mRNA in each group were detected by RT-qPCR,and expressions of PD-1 and iNOS protein in microglia were detected by Western blot.Flow cytometry was used to detect cell apoptosis in each group.Levels of inflamma-tory cytokines IL-1β,IL-6,TNF-α and IL-10 were detected by ELISA.Results:RT-qPCR and Western blot results showed that com-pared with Control group,LPS group had significantly increased expression of PD-1 and iNOS(P<0.05).Compared with LPS group,LPS+BMS-1 group had significantly decreased expression of PD-1(P<0.05)and significantly increased expression of iNOS(P<0.05).Flow cytometry showed that compared with Control group,LPS group had a significantly increased in apoptosis of microglia(P<0.000 1).Compared with LPS group,LPS+BMS-1 group had a significantly increased in apoptosis of microglia(P<0.000 1).ELISA results showed that compared with Control group,LPS group had no significantly increased in pro-inflammatory factors IL-1β and IL-6(P>0.05),while significantly increased in TNF-α(P<0.000 1)and anti-inflammatory factor IL-10(P<0.000 1).Pro-inflammatory cyto-kine IL-1β in LPS+BMS-1 group was significantly higher than that in LPS group(P=0.000 1),IL-6 and TNF-α were also significantly higher than those in LPS group(P<0.000 1),while anti-inflammatory cytokine IL-10 in LPS+BMS-1 group was significantly lower than that in LPS group(P<0.000 1).Conclusion:BMS-1 can promote LPS-induced inflammatory response or impede the recovery of inflammation,and increase apoptosis of microglia.PD-1/PD-L1 pathway may be a potential therapeutic target for neuroinflammation.

Objective:To explore the effect of Pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) with chemotherapy on breast cancer patients who got agranulocytosis of 3 to 4 degree, agranulocytosis with fever (FN) and the influential factors of relative dose intensity (RDI) chemotherapy scheme. Meantime, the value of CD34 + and CD45 in peripheral blood on predicting agranulocytosis of 3-4 degree were investigated.Methods:A total of 104 women with breast cancer were treated at Huzhou Maternal and Child Health Hospital and Cancer Hospital of Zhejiang Province from Jan. 2022 to Sep. 2023. All subjects received primary prevention with PEG-rhG-CSF during chemotherapy. The clinical risk factors of agranulocytosis, FN and RDI were analyzed. The levels of CD34 + and CD45 in peripheral blood samples were analyzed by flow cytometry. Then the predictive value of the receiver characteristic curve (ROC) for Grade 3 to 4 agranulocytosis after primary prophylaxis with PEG-rhG-CSF for breast cancer chemotherapy was evaluated.Results:Among 104 breast cancer patients who received primary prevention of PEG-rhG-CSF during chemotherapy, 28 patients had agranulocytosis of 3 to 4 grade, 10 patients got FN, and 12 patients developed RDI<85%. The results of single factor analysis showed that CD34 +, CD45 and chemotherapy scheme were the influential factors of agranulocytosis of 3 to 4 degree, and low RDI of chemotherapy scheme ( OR=0.584, OR=0.999, OR=2.299, OR=0.100, OR=0.999, OR=3.088, P<0.05) . It also showed that CD34 + and chemotherapy scheme were the influential factors of FN ( OR=0.099, OR=2.667, P<0.05) . Multivariate Logistic regression analysis showed that CD34 +, CD45 and intensive chemotherapy were the independent risk factors of agranulocytosis of 3 to 4 degree after primary prevention with PEG-rhG-CSF ( OR=0.602, OR=0.999, OR=20.174, P<0.05) . CD34 + and intensive chemotherapy scheme were the independent influential factors of FN and RDI of chemotherapy scheme after primary prevention with PEG-RHG-CSF ( OR=0.072, OR=33.934, OR=0.086, OR=54.788, P<0.05) . The area under the curve (AUC) of CD34 + were 0.767 (95% CI:0.659-0.876) , AUC of CD45 were 0.743 (95% CI:0.644-0.842) , and the AUC of combined two indexes was 0.825 (95% CI:0.730-0.920) , which was higher than that of single index. So AUC of CD34 + and CD45 can be used for predicting agranulocytosis of grade 3 to 4 in breast cancer patients receiving primary prophylaxis with PEG-rhG-CSF. Conclusions:The levels of CD34 + and CD45 in peripheral blood of breast cancer patients with agranulocytosis of grade 3 to 4 receiving primary prevention with PEG-rhG-CSF during chemotherapy are lower. Combined detection of CD34 + and CD45 in peripheral blood can predict the occurrence of agranulocytosis of grade 3 to 4 in breast cancer patients after primary prevention with PEG-rhG-CSF. Also it can provide a reliable basis for assessing the risk of grade 3 to 4 agranulocytosis.

Objective:To explore the effect of Pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) with chemotherapy on breast cancer patients who got agranulocytosis of 3 to 4 degree, agranulocytosis with fever (FN) and the influential factors of relative dose intensity (RDI) chemotherapy scheme. Meantime, the value of CD34 + and CD45 in peripheral blood on predicting agranulocytosis of 3-4 degree were investigated.Methods:A total of 104 women with breast cancer were treated at Huzhou Maternal and Child Health Hospital and Cancer Hospital of Zhejiang Province from Jan. 2022 to Sep. 2023. All subjects received primary prevention with PEG-rhG-CSF during chemotherapy. The clinical risk factors of agranulocytosis, FN and RDI were analyzed. The levels of CD34 + and CD45 in peripheral blood samples were analyzed by flow cytometry. Then the predictive value of the receiver characteristic curve (ROC) for Grade 3 to 4 agranulocytosis after primary prophylaxis with PEG-rhG-CSF for breast cancer chemotherapy was evaluated.Results:Among 104 breast cancer patients who received primary prevention of PEG-rhG-CSF during chemotherapy, 28 patients had agranulocytosis of 3 to 4 grade, 10 patients got FN, and 12 patients developed RDI<85%. The results of single factor analysis showed that CD34 +, CD45 and chemotherapy scheme were the influential factors of agranulocytosis of 3 to 4 degree, and low RDI of chemotherapy scheme ( OR=0.584, OR=0.999, OR=2.299, OR=0.100, OR=0.999, OR=3.088, P<0.05) . It also showed that CD34 + and chemotherapy scheme were the influential factors of FN ( OR=0.099, OR=2.667, P<0.05) . Multivariate Logistic regression analysis showed that CD34 +, CD45 and intensive chemotherapy were the independent risk factors of agranulocytosis of 3 to 4 degree after primary prevention with PEG-rhG-CSF ( OR=0.602, OR=0.999, OR=20.174, P<0.05) . CD34 + and intensive chemotherapy scheme were the independent influential factors of FN and RDI of chemotherapy scheme after primary prevention with PEG-RHG-CSF ( OR=0.072, OR=33.934, OR=0.086, OR=54.788, P<0.05) . The area under the curve (AUC) of CD34 + were 0.767 (95% CI:0.659-0.876) , AUC of CD45 were 0.743 (95% CI:0.644-0.842) , and the AUC of combined two indexes was 0.825 (95% CI:0.730-0.920) , which was higher than that of single index. So AUC of CD34 + and CD45 can be used for predicting agranulocytosis of grade 3 to 4 in breast cancer patients receiving primary prophylaxis with PEG-rhG-CSF. Conclusions:The levels of CD34 + and CD45 in peripheral blood of breast cancer patients with agranulocytosis of grade 3 to 4 receiving primary prevention with PEG-rhG-CSF during chemotherapy are lower. Combined detection of CD34 + and CD45 in peripheral blood can predict the occurrence of agranulocytosis of grade 3 to 4 in breast cancer patients after primary prevention with PEG-rhG-CSF. Also it can provide a reliable basis for assessing the risk of grade 3 to 4 agranulocytosis.

Objective:To investigate the effect of PD-1/PD-L1 inhibitor BMS-1 on LPS-induced inflammation in mouse microg-lia cells(BV-2 cells).Methods:Bv-2 cells were divided into Control group,LPS group and LPS+BMS-1 group.Bv-2 cells in Control group were cultured in DMEM medium for 78 hours,cells in LPS group were stimulated with 100 ng/ml LPS for 6 hours after 72 hours of normal culture,Bv-2 cells in LPS+BMS-1 group were treated with 50 nmol/ml BMS-1 for 72 hours and then stimulated with 100 ng/ml LPS for 6 hours.Expressions of PD-1 and iNOS mRNA in each group were detected by RT-qPCR,and expressions of PD-1 and iNOS protein in microglia were detected by Western blot.Flow cytometry was used to detect cell apoptosis in each group.Levels of inflamma-tory cytokines IL-1β,IL-6,TNF-α and IL-10 were detected by ELISA.Results:RT-qPCR and Western blot results showed that com-pared with Control group,LPS group had significantly increased expression of PD-1 and iNOS(P<0.05).Compared with LPS group,LPS+BMS-1 group had significantly decreased expression of PD-1(P<0.05)and significantly increased expression of iNOS(P<0.05).Flow cytometry showed that compared with Control group,LPS group had a significantly increased in apoptosis of microglia(P<0.000 1).Compared with LPS group,LPS+BMS-1 group had a significantly increased in apoptosis of microglia(P<0.000 1).ELISA results showed that compared with Control group,LPS group had no significantly increased in pro-inflammatory factors IL-1β and IL-6(P>0.05),while significantly increased in TNF-α(P<0.000 1)and anti-inflammatory factor IL-10(P<0.000 1).Pro-inflammatory cyto-kine IL-1β in LPS+BMS-1 group was significantly higher than that in LPS group(P=0.000 1),IL-6 and TNF-α were also significantly higher than those in LPS group(P<0.000 1),while anti-inflammatory cytokine IL-10 in LPS+BMS-1 group was significantly lower than that in LPS group(P<0.000 1).Conclusion:BMS-1 can promote LPS-induced inflammatory response or impede the recovery of inflammation,and increase apoptosis of microglia.PD-1/PD-L1 pathway may be a potential therapeutic target for neuroinflammation.

Objective To investigate the effect of dioscin on uric acid(UA)-induced oxidative stress injury of human renal tubular epithelial cells(HK-2)and its molecular mechanism.Methods HK-2 cells were cultured and divided into four groups:blank group(normal group),model group(uric acid-stimulation modeling),condition control group(UA+DMSO)and dioscin group(UA+dioscin).Oxidative stress injury model was induced by UA in HK-2 cells.Cells viability was detected by CCK-8.ROS level was detected by flow cytometry.Real-time PCR was used to detect the expressions of glycogen synthase kinase 3β(GSK3β),nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase 1(HO-1)at mRNA level,and Western Blot was used to detect the expressions of phosphorylated glycogen synthesis kinase 3β(p-GSK3β),GSK3β,Nrf2 and HO-1 at protein level.Results After stimulation by UA,HK-2 cells viability was obviously decreased,and ROS level was significantly increased(all P＜0.001).When treated with dioscin,HK-2 cells viability was obviously increased,and the ROS level of HK-2 cells was significantly decreased(all P＜0.001).The expressions of Nrf2 and HO-1 decreased at the protein and mRNA levels after stimulation with UA.But the expressions of Nrf2 and HO-1 significantly increased after treated with dioscin(all P＜0.001).Compared with the blank group,the p-GSK3β/GSK3β ratio in the model group decreased significantly at the protein level,but the p-GSK3β/GSK3β ratio increased after treated with dioscin(all P＜0.001).Conclusion Dioscin can alleviate UA-induced oxidative stress injury in HK-2 cells.The mechanism might be that dioscin can promote phosphorylation of GSK3β,and activate Nrf2/HO-1 pathway.

OBJECTIVE: To analyze the early effectiveness of unilateral biportal endoscopy (UBE) laminectomy in the treatment of two-level lumbar spinal stenosis (LSS). METHODS: The clinical data of 98 patients with two-level LSS treated with UBE between September 2020 and December 2021 were retrospectively analyzed. There were 53 males and 45 females with an average age of 59.9 years (range, 32-79 years). Among them, there were 56 cases of mixed spinal stenosis, 23 cases of central spinal canal stenosis, and 19 cases of nerve root canal stenosis. The duration of symptoms was 1.5- 10 years, with an average of 5.4 years. The operative segments were L _{2, 3} and L _{3, 4} in 2 cases, L _{3, 4} and L _{4, 5} in 29 cases, L _{4, 5} and L _5, S ₁ in 67 cases. All patients had different degrees of low back pain, among of which 76 cases were with unilateral lower extremity symptoms and 22 cases were with bilateral lower extremity symptoms. There were 29 cases of bilateral decompression in both segments, 63 cases of unilateral decompression in both segments, and 6 cases of unilateral decompression and bilateral decompression of each segment. The operation time, intraoperative blood loss, total incision length, hospitalization stay, ambulation time, and related complications were recorded. Visual analogue scale (VAS) score was used to assess the low back and leg pain before operation and at 3 days, 3 months after operation, and at last follow-up. The Oswestry disability index (ODI) was used to evaluate the functional recovery of lumbar spine before operation and at 3 months and last follow-up after operation. Modified MacNab criteria was used to evaluate clinical outcomes at last follow-up. Imaging examinations were performed before and after operation to measure the preservation rate of articular process, modified Pfirrmann scale, disc height (DH), lumbar lordosis angle (LLA), and cross-sectional area of the canal (CAC), and the CAC improvement rate was calculated. RESULTS: All patients underwent surgery successfully. The operation time was (106.7±25.1) minutes, the intraoperative blood loss was (67.7±14.2) mL, and the total incision length was (3.2±0.4) cm. The hospitalization stay was 8 (7, 9) days, and the ambulation time was 3 (3, 4) days. All the wounds healed by first intention. Dural tear occurred in 1 case during operation, and mild headache occurred in 1 case after operation. All patients were followed up 13-28 months with an average of 19.3 months, and there was no recurrence or reoperation during the follow-up. At last follow-up, the preservation rate of articular process was 84.7%±7.3%. The modified Pfirrmann scale and DH were significantly different from those before operation ( P<0.05), while the LLA was not significantly different from that before operation ( P=0.050). The CAC significantly improved ( P<0.05), and the CAC improvement rate was 108.1%±17.8%. The VAS scores of low back pain and leg pain and ODI at each time point after operation significantly improved when compared with those before operation, and the differences between each time points were significant ( P<0.05). According to the modified MacNab criteria, 63 cases were excellent, 25 cases were good, and 10 cases were fair, with an excellent and good rate of 89.8%. CONCLUSION UBE laminectomy is a safe and effective technique with little trauma and fast recovery for two-level LSS and the early effectiveness is satisfactory.
Male ; Female ; Humans ; Middle Aged ; Laminectomy ; Spinal Stenosis/surgery* ; Constriction, Pathologic/surgery* ; Low Back Pain ; Retrospective Studies ; Blood Loss, Surgical ; Endoscopy ; Lumbar Vertebrae/surgery* ; Spinal Fusion/methods* ; Decompression, Surgical ; Surgical Wound/surgery* ; Treatment Outcome

Objective:To examine the expression of core clock genes in the peripheral blood mononuclear cells (PBMCs) and the level of circadian disturbance-related proteins in the serum of chronic pancreatitis (CP) patients with pancreatic exocrine insufficiency (PEI), and explore their potential diagnostic value in clinical practice.Methods:The peripheral blood samples and related clinical data from 68 patients diagnosed with CP in Shanghai General Hospital from Jan 2015 to Jan 2022 were collected. Peripheral blood samples from 30 healthy individuals were used for control. The M-ANNHEIM classification system was used to stratify the clinical stages of patients with CP. The mRNA expression of the core clock genes, including Clock, Bmal1, Per1/2/3 and Cry1/2 in PBMCs was analyzed using realtime qPCR, and the expression of circadian disturbance-related proteins like TrkB, CD 36 and Rbp in serum was measured with ELISA. The receiver operating characteristic curve(ROC) and the area under curve (AUC) was used to test the efficiency for diagnozing PEI. Results:The mRNA expression of Per1 in CP patients was significantly decreased (0.76 vs 1, P<0.05), and the AUC for diagnozing PEI was 0.744 (95% CI 0.628-0.860), with a cut-off value of 0.72; and the sensitivity and specificity was 84.8% and 57.1%, respectively. The protein abundance of serum CD 36 was significantly increased in CP patients (33.85±19.74ng/ml vs 24.71±11.53 ng/ml, P<0.05); the AUC for diagnozing PEI was 0.834 (95% CI 0.735-0.932), with a cut-off value of 29.75 pg/ml; and the sensitivity and specificity was 74.3% and 84.8%, respectively. The expression of CD 36 was increased with the increase of CP clinical stage, and there were statistically significant differences between either two stages (all P value <0.05). The mRNA expression of Per1 in patients with CP in Stage Ⅰ was significantly higher than that in patients with CP in Stage Ⅱ or Ⅲ, and the differences were statistically significant ( P<0.05), but no statistical difference was found between Stage Ⅱ and Stage Ⅲ. Conclusions:The decreased expression of Per1 mRNA in PBMCs and increased level of CD 36 in serum are significantly related to the occurrence of PEI in CP, suggesting that they may have potential value for diagnozing PEI and guiding the clinical practice.

Objective To explore the therapeutic effect of radioactive 125I seeds combined with chemotherapy on the treatment of inoperative pancoast tumor by CT-guided percutaneous implantation of treatment planning system (TPS).Methods From December 2002 to December 2010,36 patients with pancoast tumor were confirmed by imaging and pathology.Among them,26 cases suffered from squamous cell carcinoma and 10 cases with adenocarcinoma.At 1 week after radioactive 125I implantation,chemotherapy was administered as intravenous 1 000 mg / m2 gemcitabine at 1 and 8 d and intravenous cisplatin 75 mg/ m2 at 1 d for 4 consecutive cycles.The prescribed dose (PD) was 120 Gy,and the median activity of the seeds was 0.7 mCi (2.59 × 107 Bq) with the range of 0.68-0.82 mCi (2.52 × 107-3.03 × 107 Bq).The patients were followed-up and the median follow-up time was 48 months.The survival rate was observed.Results The mPD of the target tumor was (118.7 ± 7.2) Gy,D90(126 ± 4.7) Gy,D90 ＞ mPD.The rate of CR (11 cases) was 63.6％,and the effective rate (CR + PR) was 83.4％.The follow-up last till December 1st,2016.1-,3-and 5-year local control rates were 92％,83％,and 67％,respectively.1-,3-and 5-year cumulative survival rate were 84.1％,56.7％,and 36.8％,respectively.Median survival was 38 months.Conclusions Chemotherapy combined with tissue radioactive 125I seed implantation is a minimally invasive and effective method for the treatment of pancoast tumor.

Objective To compare the planned radiation dose and the actual dose received after 125I radioactive seeds implantation supported by coplanar template (CPT) in lung cancer patients with mediastinal node metastases 4R,and to evaluate the clinical efficacy.Methods Totally 32 patients with lung cancer with mediastinal lymph node metastases 4R who had been diagnosed via cellular pathology studies were selected from January 2008 to December 2014.The mediastinal lymph node metastases were treated by CPT-assisted 125I radioactive seed implantation.Digital imaging and communications in medicine (DICOM) data were acquired by chest CT scan before implantation,brachytherapy radiation treatment planning system (BTPS) was introduced to carry out the plan,and the prescribed dose(PD) was 120 Gy.CPT was used to control the precision of needle penetration and implantation of radioactive seeds.Computer tomography (CT) was used to ensure the correct position of needles and radioactive seeds.Dose evaluation was performed immediately after implantation and was compared with planned dose using paired t-test.The 6-months postoperative chest CT was conducted to evaluate treatment efficacy according to response evaluation criteria in solid tumors(RECIST Version 1.1).Results All patients went through implantation procedure successfully.Dose evaluation after implantation was as followed.The average dose received (231.9 ±29.6)Gy,the dose received by 90％ of the target(D90) (150.8 ± 16.6) Gy,the dose received by 100％ of the target(D100) (100.4 ± 12.6)Gy,the volume of 100％PD covering the target(V100)(94.1 ± 2.6) ％,the volume of 200％ PD covering the target(V200) (33.0 ± 5.7) ％,the conformal index (CI) 0.75 ±0.06,the external index(EI) (22.7 ± 5.8)％,the average dose received by the superior vena cava (19.3 ± 7.2)Gy,and the average dose received by aorta (12.1 ± 5.1)Gy.Efficacy was followed for 6 months after implantation and the effective rate was 84.37％.There was no serious complications (such as radioactive lung injury,major vascular injury,bleeding,and et al.) occurred in follow-up period.Conclusions CPT assisted CT guided 125I radioactive seed imnplantation in treating mediastinal node metastases 4R can achieve preoperative BTPS,minimize major vascular or organ injury.It is an accurate,effective and safe treatment approach and may be of great value to standardize the procedure of radioactive seed implantation in mediastinal metastases.

Objective To investigate the feasibility of low dose dual phase contrast-enhanced chest CT with iterative mod el reconstruction (IMR) technique.Methods Totally 130 patients with suspected pulmonary occupying lesions underwent dual phase contrast-enhanced chest CT,who were randomly assigned into 2 groups (group A and group B,each n=65).Patients in group A were scanned with 100 kV,DoseRight technique with dose right index 10,and images were reconstructed with the hybrid iterative reconstruction (iDose4).While patients in group B were scanned with 80 kV,DoseRight technique with dose right index 8,and images were reconstructed with iterative model reconstruction (IMR).The objective image quality,subjective image scores and the excellence rate of vascular visualization were compared in both pulmonary artery (PA) and bronchial artery (BA) phases.The radiation dose was also calculated.Results The effective dose was (3.30 ±0.89)mSv in group A and (1.27 ±0.19)mSv in group B.Compared to group A,the effective dose reduced 61.52％ in group B (P＜0.001).Lower image noise and greater CNR were obtained in group B compared to group A in both PA and BA phases (all P＜0.001).No significant difference was found in subjective image scores of lung and mediastinal setting and the excellence rate of vascular visualization in both groups (all P＞0.05).Conclusion Using IMR,dual phase contrast-enhanced chest CT allows for a radiation dose reduction up to 61.52％,meanwhile,ensures the image quality and meets the diagnostic requirements.