Objective To establish a quantitative method for Astraloside in Xingnao Zaizao Capsules by HPLC-ELSD. Methods HPLC-ELSD separation was achieved on Phenomenex Gemini C18 column (250 mm?4.60 mm, 5 ?m) and mobile phase consisted of acetonitrile-water (36∶64). The flow rate was 1.0 mL/min, drift tube temperature was 105 ℃, and gas flow rate was 2.7 L/min. Results The calibration curve was linear in the range of 0.486～2.43 ?g (r=0.999 7). The average recovery rate was 100.5% (n=6), RSD was 0.3%. Conclusion The method is rapid, accurate and reliable, and the method can be used as the quality standard for Xingnao Zaizao Capsules.

Objective To observe learning and memory behavior changes of one time cerebral concussion called pure cerebral concussion(PCC)and three times cerebral concussion called multiple cerebral concussion (MCC)after injured 24 days in rats.Methods A metallic pendulum striker device was deployed to duplicate PCC and MCC model in SD rata which were the complete closed head injury model.The animals were divided into PCC and MCC groups at random.One control group was used,each group has eight animals(n=8).One 8-arms radial maze was used to assessed each animal's capabilities,that is,spatial reference memory,working memory,spirited activity and take in food.Results Compared with control group,there were some significance(P＜0.05)in both experiment groups post injury,that was,(1)The food intake decreased,PCC group from the 1st to the 11th day(from 0.00±0.00 to 2.62±1.76)after injury,MCC group from the 1st day to the 24th day(from 0.00±0.00 to 0.75±1.48)after injury.(2)Spirited activity depressed,PCC group on the 1st to the 7th,13rd day(from 4.87±1.24 to 10.0±2.39)after injury,MCC group on the 1st to 8th,22nd day(from 4.25±5.03 to 9.37±4.20)after injury.(3)The spatial reference memory was lower in early then gradually increased,PCC group on the 1 st to 7th day(from 0.50±0.75 to 3.O0±1.06)after injury,MCC group from the 1st to 19th day(from 1.88±2.10 to 2.50±2.44)after injury.(4)The working memory was delaying damaged,PCC group from the 1st to the 6th day and the 10th to the 23rd day(from 0.00±0.00 to 4.25±3.05)after injury,MCC group on the 1～4th,6th,9～13th,15th,16th,19th～22nd day(from 0.25±0.46 to 3.12±2.87)after injury.Conclusion The injured rata'capability of spatial reference memory,working memory,spirited activity and food intake were obviously damaged after CC,and the MCC group's capability of spatial reference memory,spirited activity and food intake was worse than PCC group.

Objective To evaluate the significance of contrast medium (CM) volume and estimated glomerular filtration rate (CM/eGFR) in predicting contrast-induced nephropathy (CIN) after PCI. Methods A total of 307 patients after PCI were enrolled from Nanfang Hospital from May 2014 to October 2015. The patients were divided into the CIN group(n = 29) and the non-CIN group(n = 278) according to whether CIN within 72 hours after PCI. The baseline renal function was assessed by the sCr and CyC, respectively. Results Twenty-nine patients (9.4%, 29/307) developed CIN. There were significant differences in Age, CM、NTpro-BNP、IABP、 Periprocedural Hypotension、Preprocedural sCr/CyC between two groups (P < 0.05, respectively). The result of multivariate logistic regression analysis showed that Age, Cardiac function ≥Ⅲ level, IABP, use CCB, CM/eGFRMDRD, CM/eGFRCyC were independent risk predictors for CIN, respectively. Receiver Operating Characteristic (ROC) curve analysis showed that the area under the curve of CM/eGFRMDRD(AUC = 0.838) was superior to CM/eGFRCyC (AUC = 0.805) without significant difference. The sensitivity and specificity were 79.3%and 76.3%(Cut-off Point = 2.094), respectively. Conclusion Both the CM/eGFRMDRD and CM/eGFRCyC may be good methods to determine maximum CM before PCI and to predict CIN after PCI currently, without significant differences between these two predictors.

Objective To evaluate the prognostic significance of carbonic anhydrase IX (CA IX) expression in patients with clear cell renal cell carcinoma (ccRCC). Methods CA IX expression in a cohort of 120 patients with ccRCC was evaluated by P-V immunohistochemistry with a rabbit CA IX polyclonal antibody. Twenty-five normal kidney tissues were used as a control. The relationship between CA IX expression and prognosis was analyzed by univariate and multiple-factor analysis (Cox regression model). The primary end point was cancer specific survival. Results One hundred and twelve (93.3%) patients were followed up with the median follow-up time of 45 months (range, 6 to 94 months). Seventy-five patients survived without evidence of tumor recurrence, 3 patients survived with tumor recurrence, and 34 patients died, 28 of the 34 died of cancer. CA IX expression was negative in all normal renal tissue. High CA IX expression was observed in 89 (74.2%) patients, among which 82 patients were followed up, and the disease free survival was 75.6% (62/82). Two (2.4%) patients survived with tumor recurrence, and 18 (22.0%) patients died, of which 13 (15.9%) died of cancer. Tumor recurrence and (or) metastasis occurred in 9 (11.0%) patients, with a median survival of 92 months in this high expression group. Low CA IX expression was observed in 31 (25.8%) patients, among which 30 patients were followed up, and the disease free survival was 43.3% (13/30). One (3.3%) patient survived with tumor recurrence, and 16 (53.3%) patients died, of which 15 (50.0%) died of cancer. Tumor recurrence and (or) metastasis occurred in 8 (26.7%) patients with a median survival of 53 months in this low expression group. Cancer specific survival between CA IX high expression group and low expression group was significantly different (P=0.000, χ2=15.950), and tumor relapse and (or) metastasis rates were significantly different (P=0.040, χ2=4.200). The 1, 3, 5 and 7 year cancer specific survival rates were 95.2%, 83.9%, 81.2% and 78.2% respectively in the high CA IX expression group, and 89.5%, 63.9%, 46.8% and 40.1% respectively in the low expression group. Multivariate analysis with Cox regression model showed that CA IX expression was a prognostic factor (RR=0.186). Conclusions High CA IX expression is negatively correlated with postoperative mortality, relapse and (or) metastasis in ccRCC. CA IX expression could be used as a prognostic biomarker in ccRCC.

Objective: To observe the changes of PD-1 expression, mRNA level and cytotoxic activity of CD19 CAR-T cells during the culture process of CAR-T cells. Methods: The peripheral blood T cells of 6 lymphoma patients with high expression of PD-1 and 6 healthy volunteers were the source of CAR-T cells. The expression of PD-1 was analyzed by flow cytometry. The mRNA level of PD-1 was analyzed by PCR. The cell proliferation was analyzed by CCK-8 assay. The cytotoxicity was analyzed by LDH assay. Results: ①The transfection efficiency of high PD-1 expression T cells and healthy volunteer T cells were as the same (P>0.05) . ②The cell proliferation capacity of CD19 CAR-T cells from high PD-1 expression T cells or healthy volunteer T cells, with or without PD-1 inhibitor were as the same (P>0.05) . ③The cytotoxicity to lymphoma cells of high PD-1 expression T cells and CAR-T cells were lower than that of these two T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer T cells (P<0.001) . There was no difference of the cytotoxicity between the CAR-T cells from high PD-1 expression T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer (P>0.05) . ④There was no difference of the expression of PD-1 in all CAR-T cell groups during the culture process (P>0.05) . There was no difference of mRNA level of PD-1 in all groups during the culture process (P>0.05) . ⑤The PD-1 expression of CAR-T cells increased by the time of culture after contacting with lymphoma cells (P<0.001) . The PD-1 inhibitors could antagonize this effect. There was no difference of mRNA level of PD-1 in all groups after contacting with lymphoma cells (P>0.05) . Conclusion The PD-1 expression of CAR-T cells from high PD-1 expression T cells increased by the time of culture after contacting with lymphoma cells. However, the mRNA level of PD-1 of all groups did not change, even if PD-1 inhibitor was applied.

Objective: To Evaluation the effect of PD-1 inhibitor Nivolumab on the proliferation and cytotoxicity of anti-CD19 chimeric antigen receptor T cells (CD19-CAR-T) in vitro. Methods: Five patients with high PD-1 expression in peripheral blood and five healthy volunteers were selected. These peripheral blood mononuclear cells were used as the source of T cells to prepare CD19-CAR-T cells. Different doses (72, 36, 18 μg/ml) of Nivolumab was added on day 8 to the culture medium. Patient T cells incubated with 72 μg/ml Nivolumab and CD19-CAR-T cells of healthy volunteers were used as controls. CCK-8, lactate dehydrogenase (LDH) cytotoxicity assay and ELASA were used to detect the proliferation capacity, the specific cytotoxicity and the inflammatory factor secretion. Results: ①T cells from patients with high expression of PD-1 as the source of CD19-CAR-T cells did not affect transfection rate compared with that of healthy volunteers [(32.80±7.22)% vs (35.10±5.84)%, t=-0.554, P=0.593]. ②Incubation of CD19-CAR-T cells with 72 μg/ml Nivolumab did not affect CD19-CAR-T cell proliferation, but its cytotoxicity was significantly higher than that of CD19-CAR-T cells alone or patients’ T cells +72 μg/ml Nivolumab (all P<0.001), there was no significant difference in the killing activity between the 72 μg/ml and 36 μg/ml Nivolumab treated CD19-CAR-T cells on Pfeiffer cells (P=0.281, 0.267, respectively), and they were all higher than those of 18 μg/ml Nivolumab treated CD19-CAR-T cells (all P<0.001). ③Different doses of PD-1 inhibitor Nivolumab combined with CD19-CAR-T cells does not affect the secretion of IFN-γ and IFN-α (all P>0.05). Conclusion Combination of 36 μg/ml PD-1 inhibitor and CD19-CAR-T cells could reduce the drug toxicity and enhance the cytotoxicity.

Objective:To compare the clinical effect of transpedicular impaction and grafting of allogeneic bone containing enriched bone marrow combined with posterior internal fixation and posterior subtotal vertebrectomy combined with posterior internal fixation in the treatment of stage III Kümmell′s disease.Methods:A retrospective cohort study was made on clinical data of 40 patients with stage III Kümmell′s disease admitted to Zhengzhou Orthopedic Hospital from June 2015 to December 2018. There were 10 males and 30 females, at age range of 57-79 years［(67.7±6.1)years］. A total of 19 patients were treated by transpedicular impaction and grafting of allogeneic bone containing enriched bone marrow combined with posterior internal fixation (impaction bone graft group), and 21 patients by posterior subtotal vertebrectomy combined with posterior internal fixation (subtotal vertebrectomy group). Operation time and intraoperative blood loss were compared between the two groups. Degree of pain, lumbar dysfunction and degree of kyphosis were evaluated by visual analogue scale (VAS), Japanese Orthopedic Association (JOA) score and kyphotic Cobb angle before operation, at 1 week after operation and at the last follow-up. Bone healing time was compared between the two groups. The complications of the two groups were observed.Results:All patients were followed up for 25-64 months［(40.6±10.4)months］. Operation time and intraoperative blood loss were (130.0±10.1)minutes and (284.5±43.5)ml in impaction bone graft group, lower than those in subtotal vertebrectomy group［(253.8±33.2)minutes, (889.1±95.7)ml］(both P<0.01). There were no significant differences in VAS, JOA score or kyphotic Cobb angle between the two groups before operation, at 1 week after operation and at the last follow-up (all P>0.05). Both VAS and JOA score showed significant differences within each group at any time point (all P<0.01). In both groups, the kyphotic Cobb angle reduced significantly at 1 week after operation when compared with that before operation (all P<0.01), and the angle showed a slight increase at the last follow-up, but remained significantly lower than that before operation (all P<0.01). There were no relapse of pain or aggravation of kyphosis. Bone healing time in impaction bone graft group［4.4(4.0, 5.0)months］was significantly shorter than that in subtotal vertebrectomy group［6.4(5.2, 8.1)months］( P<0.01). There were 2 patients with delayed healing of surgical incision in impaction bone graft group, with the complication rate of 11%. There were 2 patients with dural tear and 3 patients with delayed healing of surgical incision in subtotal vertebrectomy group, with the complication rate of 24%. The complication rate was not statistically significant between the two groups ( P>0.05). No loosening or breakage of internal fixation was observed during the follow-up. Conclusions:Transpedicular impaction and grafting of allogeneic bone containing enriched bone marrow combined with posterior internal fixation and posterior subtotal vertebrectomy combined with posterior internal fixation are effective in the treatment of stage III Kümmell disease. However, the former can shorten the operation time, reduce the intraoperative blood loss and accelerate the healing of injured vertebral bone, suggesting a relatively minimally invasive surgical method for reconstruction and maintenance of spinal biomechanical stability.

Objective:To evaluate the safety and efficacy of one-stage posterior-only jumping hemivertebra (HV) resection combined with respective short fusions in the treatment of congenital scoliosis (CS) caused by multiple HVs.Methods:All of 13 consecutive patients with multiple HVs treated surgically from January 2010 to December 2017 were retrospectively reviewed, including 4 males and 9 females with a mean age of 3.7±1.2 years. One child had 4 HVs, and the rest had 2 HVs. The responsible HVs causing local scoliosis/kyphosis deformity or coronal plane deviation were selected as the target of resection. The distal HV was removed firstly and then the proximal one was resected; both of the fixation vertebraes were horizontalized during surgery. The clinical and imaging data of the children before the initial operation, immediately after the operation and at the latest follow-up were collected, and the short-term and long-term complications related to surgery were recorded. The data were evaluated on the whole-standing spine anteroposterior and lateral films, including the corrections of proximal and distal main curves, coronal balance, local kyphosis, and the improvement of spinal growth height (upper and lower internal fixation length, T 1-S 1 length). At the same time, the re-progression of coronal and sagittal deformities of the spine during growth was recorded (coronal decompensation: emerging postoperative curve progression more than 20°; kyphosis progression: kyphosis aggravation between upper and lower internal fixation more than 40°) and internal-fixation-related complications (screw cutting, screw malposition) were recorded. Results:Dual HVs were resected in each child, of which 8 (61.5%) were located on contralateral side of the spine, and 5 (38.5%) were located on ipsilateral side of the spine. The follow-up time was 6.2±3.3 years (range 2.0-10.5 years) after surgery. The Cobb angles of proximal and distal main curves were 36.7°±11.8° and 35.2°±7.8° respectively before surgery and were corrected to 9.7°±6.6° and 6.1°±4.1° respectively after surgery ( F=31.249, F=93.83, P< 0.001) ( t=6.888, t=10.954, P<0.001), and the correction rates was 73.6%±19.6% and 82.7%±11.7%, respectively. They were maintained at 14.3°±5.4° and 8.0°±4.6° at the latest follow-up, showing the correction rates loss of 15.8%±26.9% and 6.9%±7%, respectively. The coronal balance improved from 17.2±14.8 mm pre-operatively to -0.2±15.7 mm postoperatively ( t=2.703, P=0.008), and it remained at 0±18.4 mm at the final follow-up ( F=4.137, P=0.024). The T 1-S 1 length was corrected to 273.8±27.3 mm postoperatively, slightly increased compared with pre-operation 256.3±24.0 mm, ( t=0.680, P=0.527), and significantly increased to 333.2±33.4 mm at the latest follow-up ( t=2.986, P<0.001; F=6.704, P=0.003). Seven patients had local kyphosis before operation, which was significantly improved from 32.2°±13.6° to 6.1°±9.8° with a correction rate of 93.4%±27.0% after surgery ( t=3.355, P=0.004), which showed no significant loss of correction at the latest follow-up (5.4°±10.4°) ( F=11.187, P=0.002). Six patients (46.2%) developed coronal decompensation (Curve magnitude >20 °), with an average of 21.7°±1.9°. Two cases (15.4%) had progressive kyphosis between the thoracic regional internal fixations at 3 months after surgery, which were 68° and 58° respectively. After bracing, both coronal decompensation and sagittal kyphosis were improved. At the last follow-up, the coronal decompensation was improved to 14.7±8.9° and the kyphosis was alleviated to 55° and 46°, respectively. Conclusion:Posterior-only skipping hemivertebra resection and short fusion is a safe, effective procedure yielding significantly improvement of the growth imbalance and reginal spinal deformities of CS with multiple HVs. The mid-term follow-up results showed that the progress of the scoliosis was common during the growth period, which could be further controlled by supplementary brace treatment.

BACKGROUND:Stage Ⅲ Kümmell's disease is characterized by a high degree of vertebral compression and posterior wall defects.Most of the patients are elderly people with severe osteoporosis and various medical diseases.Clinically,some surgical methods are often at high risk and are controversial. OBJECTIVE:To investigate the clinical efficacy of screw placement combined with transpedicular impaction bone grafting in the treatment of stage Ⅲ Kümmell's disease. METHODS:The clinical data of injured vertebral screw placement combined with transpedicular impaction bone grafting in treatment of stage Ⅲ Kummell's disease from May 2016 to August 2021 were retrospectively analyzed.Visual analog scale score,Oswestry disability index,anterior vertebral heights,kyphotic Cobb angle and American Spinal Injury Association(ASIA)impairment scale were used to evaluate the effects of surgery.The operation time,intraoperative blood loss and complications were recorded.CT scans were used to evaluate the healing of injured vertebrae at the final follow-up visit. RESULTS AND CONCLUSION:(1)A total of 26 patients were included,with 7 males and 19 females,at the age range of 62-81 years[mean(69.7±4.8)years].The follow-up time was 18-60 months[mean(35.1±8.9)months].The average operative duration was 133.5 minutes(100-165 minutes),and the average intraoperative blood loss was 285.3 mL(210-350 mL).(2)Visual analog scale and Oswestry disability index scores 1 week after surgery were significantly lower than those before surgery.(3)At 1 week after surgery,the anterior vertebral height corrections and the Cobb angle were(9.0±0.7)mm and(16.2±1.0)°,respectively.During the follow-up period,the loss of vertebral height and kyphosis correction were(5.1±0.3)mm and(8.0±0.4)°,respectively.(4)14 patients(54%)had ASIA grade D before operation,which recovered to grade E at the last follow-up.CT scan showed that all patients achieved good osseous union.(5)Complications occurred in seven patients(27%),including hypostatic pneumonia in two cases,postoperative superficial wound tissue liquefaction in two cases,and adjacent vertebral compression fractures in three cases.(6)It is concluded that screw placement of the injured vertebra combined with transpedicular impaction bone grafting can rapidly rebuild spinal stability,effectively relieve pain and improve neurological function in the treatment of stage Ⅲ Kümmell's disease.This technique is an effective and relatively minimally invasive surgical option.

Objective:To compare the efficacies of posterior long segment instrumentation combined with transpedicular impaction bone grafting or with bone cement augmentation in treating stage III Kümmell disease.Methods:A retrospective cohort study was conducted to analyze the clinical data of 38 patients with stage III Kümmell disease who were admitted to Zhengzhou Orthopedic Hospital between January 2016 and December 2020. The study included 8 male and 30 female patients, with ages ranging from 59 to 81 years [(68.9±4.9)years]. The vertebral fractures occurred at T 8 in 1 patient, T 11 in 9 patients, T 12 in 10 patients, and L 2 in 10 patients. Seventeen patients underwent posterior long segment instrumentation combined with transpedicular impaction bone grafting (impaction bone grafting group), and 21 patients underwent posterior long segment instrumentation combined with bone cement augmentation (bone cement group). The surgical duration, intraoperative blood loss, and incidences of postoperative complications were compared between the two groups. Additionally, the visual analogue score (VAS), Japanese orthopedic association (JOA) score, and Cobb angle were compared before the operation, at 1 week and 3 months post-operation, and at the final follow-up for both groups. The study also compared bone healing at the last follow-up and postoperative complication rates between the two groups. Results:All the patients were followed up for 24-35 months [(28.7±2.9)months]. The impaction bone grafting group had a surgical duration of (150.7±25.4)minutes and intraoperative blood loss of (285.3±48.6)ml, significantly different from those in the bone cement group [(132.0±21.1)minutes, (251.4±44.8)ml] (all P<0.05). Before the operation, there were no significant differences in the VAS, JOA score, or Cobb angle between the two groups (all P>0.05).The VAS was (3.2±0.8)points, (2.7±0.5)points and (2.2±0.7)points in the impaction bone grafting group and was (2.7±0.6)points, (2.6±0.7)points and (2.4±0.8)points in the bone cement group at 1 week and 3 months post-operation and at the final follow-up, respectively. The VAS in the impaction bone grafting group was significantly higher than that in the bone cement group at 1 week post-operation ( P<0.05); however, no significant differences were found at 3 months post-operation or at the last follow-up (all P>0.05). There was no significant difference in the JOA score between the two groups at 1 week or 3 months post-operation, or at the final follow-up (all P>0.05). The Cobb angle in the impaction bone grafting group was (5.1±1.3)°, (5.9±1.8)° and (6.5±2.5)° at 1 week and 3 months post-operation, and at the final follow-up, significantly lower than that in the bone cement group [(8.4±1.6)°, (12.6±2.1)°, and (14.5±3.3)°] (all P<0.01). All the patients in the impaction bone grafting group achieved bone healing at the last follow-up. One patient in the impaction bone grafting group experienced delayed incision healing, whereas two patients in the bone cement group had poor bone healing. The complication rate was 5.9% (1/17) in the impaction bone grafting group and 9.5% (2/21) in the bone cement group ( P>0.05). Conclusions:Posterior long segment instrumentation combined with transpedicular impaction bone grafting or with bone cement augmentation are both effective in alleviating pain and improving the spinal function for stage III Kümmell disease. The former procedure is associated with longer surgical duration and increased intraoperative blood loss, but it can provide superior correction and maintenance of kyphosis deformity, promoting the healing of the injured vertebrae.