Objective To establish the quality standard of Fufang Tianma Gouteng Orally Disintegrating Tablets. Methods Gastrodiae Rhizoma, Uncariae Ramulus Cum Uncis, Angelicae Sinensis Radix and Paeoniae Radix Alba in the preparation were identified by TLC, the content of gastrodin in gastrodia was determined by HPLC. Results The herbs of preparation could be identified by TLC without interference. Gastrodin showed a good linear relationship in the range of 0.191 2-1.147 2 μg (r=0.999 9). The average recovery was 97.72% (RSD=1.40%, n=6). Conclusion The methods are available with a good reproducibility, and can control the quality of the preparation effectively.

Objective To investigate the clinical characteristics,surgical experience and mid-term outcome of coronary artery bypass graft surgery in patients aged ≤ 40 years.Methods From Jan.,2009 to Dec.,2015,12 patients with median age of (37.9 ± 2.5) years (range 32-40 years,10 males) were diagnosed with coronary artery disease and underwent coronary artery bypass graft surgery at Zhongshan Hospital,Fudan University.The clinical characteristics included five cases with hypertension,three cases with diabetes mellitus,four cases with hyperlipemia,eight cases with smoking history,seven cases with myocardial infarction.Six cases had at least one branch totally occluded.Two cases once underwent percutaneous coronary intervention surgery and implanted with five and three stents respectively.The perioperative data and follow-up results were retrospectively analysed.Results There was no in-hospital death and no death during follow-up period.Surgical techniques included bilateral internal mammary arteries combined with radial artery were operated in six cases,bilateral internal mammary arteries combined with great saphenous vein in two cases,left internal mammary arteries combined with great saphenous vein in four cases,off-pump in nine cases and on-pump in three cases.The mean follow-up time was (47.8 ± 24.3) months.During the follow-up one case suffered with saphenous vein graft restenosis 5 years after the surgery and underwent percutaneous coronary intervention.Other cases were all alive and assessed New York Heart Association's function class Ⅰ-Ⅱ.Conclusions Younger patients usually suffer with severe coronary artery disease when diagnosed because they often have excellent compensative capacity and the pathogenesis is concealed.The mid-term results of coronary artery bypass graft surgery in patients aged ≤40 years are satisfactory.

Objective To investigate the role of holoenzyme containing Protein Phosphatase 2A B56βin regulating CdCl2 induced cytotoxicity. Method CdCl2-induced cytotoxicity in normal human cell line L-02,AFB1-transformed hepatic cell line L-02 RT-AFB1 and tumor cell line Bel7402 was measured by modified MTT assay. Stable cell lines L-02 SHAKT, L-02 SHB56β, L-02 RT-AFB1-B56β and Bel7402-B56β were generated by infecting L-02 cells or Bel7402 cells with retroviral vectors encoding lentiviral AKT shRNA, lentiviral B56βshRNA and B56β. The relative cell viability was measured in normal human cell line AFB1-transformed hepatic cell line and tumor cell line when treated by CdCl2(0,20,40,80, 160 μmol/L). After treated by wortmannin (2.5,5.0 μmol/L) combined with 40 μmol/L CdCl2,Western blot was applied to measure the expression of associated protein in L-02.Western blot was applied to measure the expression of B56β, MT (metallothionein), AKT, and p-AKT in these cell lines treated by CdCl2. Results The levels of MT were 0.12 ± 0.02, 0.06 ± 0.06 in L-02 RT-AFB1 and Bel7402,which were lower than L02 (0.92 ± 0.14) (F=1 148.16,P<0.001)when treated by 40μmol/L CdCl2.When treated by 40μmol/L CdCl2,the expression of p-AKT in L-02 SHAKT-1 and L-02 SHAKT-2 were 0.08 ± 0.02, 0.08 ± 0.05,which levels were lower than L-02 SHGFP(0.18 ± 0.15) (F=724.70,P<0.001);and the expression of MT were both 0.62 ± 0.16 in L-02 SHAKT-1 and L-02 SHAKT-2,which levels were higher than L-02 SHGFP (0.22 ± 0.14) (F=94.73,P<0.001).After treated by wortmannin (2.5,5.0 μmol/L) combined with 40 μmol/L CdCl2, the expression of p-AKT in L-02 were 0.28±0.07, 0.15±0.11,which levels were lower than wortmannin untreated cells (0.52± 0.11) (F=578.57,P<0.001);and the expreesion of MT were 1.62 ± 0.80, 1.08 ± 0.15,which levels were higher than wortmannin untreated cells (0.69 ± 0.18) (F=12.34,P<0.001).When treated by 40 μmol/L CdCl2,the levels of p-AKT in L-02 SHB56β-1 and L-02 SHB56β-2 were 0.57±0.13, 0.59±0.02,which were higher than L-02 SHGFP(0.32 ± 0.02) (F=87.16,P<0.001); and the levels of MT were 0.35 ± 0.07, 0.20 ± 0.03 in L-02 SHB56β-1 and L-02 SHB56β-2,which were lower than L-02 SHGFP (1.51 ± 0.13) (F=2 457.10,P<0.001). After treated by 40μmol/L CdCl2, the expression of p-AKT in L-02 RT-AFB1-B56βand Bel7402-B56βwere 0.10 ± 0.11, 0.09 ± 0.01,which were lower than L-02 RT-AFB1 (0.36 ± 0.01) and Bel7402 (0.43 ± 0.11) (F=877.62,P<0.001); and the levels of MT were 0.92 ± 0.13, 0.95 ± 0.08 in L-02 RT-AFB1-B56β and Bel7402-B56β,which were higher than L-02 RT-AFB1 (0.44 ± 0.12) and Bel7402 (0.77 ± 0.06) (F=51.97,P<0.001). Conclusion Protein phosphatase 2A complexes containing B56βparticipated in the regulation of MT expression through direct dephosphorylation of AKT, finally affected the cytotoxicity responding to CdCl2. Our study revealed a key signaling pathways of PP2A involved in heavy metals induced cytotoxicity.

Objective To investigate the role of holoenzyme containing Protein Phosphatase 2A B56βin regulating CdCl2 induced cytotoxicity. Method CdCl2-induced cytotoxicity in normal human cell line L-02,AFB1-transformed hepatic cell line L-02 RT-AFB1 and tumor cell line Bel7402 was measured by modified MTT assay. Stable cell lines L-02 SHAKT, L-02 SHB56β, L-02 RT-AFB1-B56β and Bel7402-B56β were generated by infecting L-02 cells or Bel7402 cells with retroviral vectors encoding lentiviral AKT shRNA, lentiviral B56βshRNA and B56β. The relative cell viability was measured in normal human cell line AFB1-transformed hepatic cell line and tumor cell line when treated by CdCl2(0,20,40,80, 160 μmol/L). After treated by wortmannin (2.5,5.0 μmol/L) combined with 40 μmol/L CdCl2,Western blot was applied to measure the expression of associated protein in L-02.Western blot was applied to measure the expression of B56β, MT (metallothionein), AKT, and p-AKT in these cell lines treated by CdCl2. Results The levels of MT were 0.12 ± 0.02, 0.06 ± 0.06 in L-02 RT-AFB1 and Bel7402,which were lower than L02 (0.92 ± 0.14) (F=1 148.16,P<0.001)when treated by 40μmol/L CdCl2.When treated by 40μmol/L CdCl2,the expression of p-AKT in L-02 SHAKT-1 and L-02 SHAKT-2 were 0.08 ± 0.02, 0.08 ± 0.05,which levels were lower than L-02 SHGFP(0.18 ± 0.15) (F=724.70,P<0.001);and the expression of MT were both 0.62 ± 0.16 in L-02 SHAKT-1 and L-02 SHAKT-2,which levels were higher than L-02 SHGFP (0.22 ± 0.14) (F=94.73,P<0.001).After treated by wortmannin (2.5,5.0 μmol/L) combined with 40 μmol/L CdCl2, the expression of p-AKT in L-02 were 0.28±0.07, 0.15±0.11,which levels were lower than wortmannin untreated cells (0.52± 0.11) (F=578.57,P<0.001);and the expreesion of MT were 1.62 ± 0.80, 1.08 ± 0.15,which levels were higher than wortmannin untreated cells (0.69 ± 0.18) (F=12.34,P<0.001).When treated by 40 μmol/L CdCl2,the levels of p-AKT in L-02 SHB56β-1 and L-02 SHB56β-2 were 0.57±0.13, 0.59±0.02,which were higher than L-02 SHGFP(0.32 ± 0.02) (F=87.16,P<0.001); and the levels of MT were 0.35 ± 0.07, 0.20 ± 0.03 in L-02 SHB56β-1 and L-02 SHB56β-2,which were lower than L-02 SHGFP (1.51 ± 0.13) (F=2 457.10,P<0.001). After treated by 40μmol/L CdCl2, the expression of p-AKT in L-02 RT-AFB1-B56βand Bel7402-B56βwere 0.10 ± 0.11, 0.09 ± 0.01,which were lower than L-02 RT-AFB1 (0.36 ± 0.01) and Bel7402 (0.43 ± 0.11) (F=877.62,P<0.001); and the levels of MT were 0.92 ± 0.13, 0.95 ± 0.08 in L-02 RT-AFB1-B56β and Bel7402-B56β,which were higher than L-02 RT-AFB1 (0.44 ± 0.12) and Bel7402 (0.77 ± 0.06) (F=51.97,P<0.001). Conclusion Protein phosphatase 2A complexes containing B56βparticipated in the regulation of MT expression through direct dephosphorylation of AKT, finally affected the cytotoxicity responding to CdCl2. Our study revealed a key signaling pathways of PP2A involved in heavy metals induced cytotoxicity.

Objective To investigate the effect of injection volume on clinical efficacy after minimally invasive puncture and drainage in the treatment of moderate basal ganglia cerebral hemorrhage underlying controlling the dosage of drugs in the elderly.Methods After successfully inserting the puncture needle into the center point in the largest layer long axis,the liquid hematoma was aspirated,and 40,000 units urokinase was injected into the hematoma cavity twice a day(2 mL in the small volume Group(SG),and 4 mL in the large volume group(LG),respectively).The needle were then adjusted and removed according to the hematoma clearance.The clearance rate of hematoma,intracranial pressure,edema volume,mortality,incidence of complications,neurological deficit score(NDS)and activities of daily living(ADL)were compared and analyzed.Results There were no significant differences in the first hematoma clearance rate,postoperative ICP and mortality between the two groups(P>0.05).Comparing to the SG,there was a relative higher postoperative hematoma clearance rate and lower volume of perihematoma edema in the LG(P<0.05).Meanwhile,the needle retention time,urokinase usage and cumulative complications at 28 days after operation were lower than in the SG(P<0.05).For two parameters representing long-term outcome,the NDS and ADL scores were much better in LG than in SG(P<0.05).Conclusion Large volume injection can effectively drain the hematoma,shorten the needle retention time,reduce the usage of urokinase,decrease the incidence of postoperative complications and improve the prognosis of patients,thereby providing effective and safe method in treating moderate basal ganglia cerebral hemorrhage in the elderly.

Objective:To investigate the efficacy and safety of separated R-CHOP in older patients with newly diagnosed diffuse large B-cell lymphoma(DLBCL).Methods:A total of 137 patients aged 65-80 years newly diagnosed with DLBCL between April 2013 and September 2022 at The First Affiliated Hospital of Zhengzhou University were enrolled.The patients were assigned into separated R-CHOP,full-dose R-CHOP,and reduced R-CHOP-like groups based on their different chemotherapy regimens.All individuals were treated in 21-day cycles for 4-8 cycles.The short-term and long-term efficacies and adverse reactions of the treatments were compared among the three groups,and factors influencing progression-free survival(PFS)and overall survival(OS)were analyzed.Results:The overall response rates(ORR)of patients in the separated R-CHOP,full-dose R-CHOP,and reduced R-CHOP-like groups were 89.7%,90.3%,and 86.1%,respectively,with no significant differences among them.The complete respond rate(CRR)of the separated R-CHOP group(64.1%)was significantly higher than that of the reduced R-CHOP-like group(33.3%)(P=0.008)but not significantly different from that of the full-dose R-CHOP group(66.1%).Survival curve analysis revealed no significant differences in PFS and OS between the separated and full-dose R-CHOP groups.Although the separated R-CHOP group showed improved PFS compared with the reduced R-CHOP-like group(P=0.036),there was no statistical difference in OS between these two groups.Multivariate analysis revealed that the international prognostic index(IPI)and separated R-CHOP had significant effects on PFS in patients with DLBCL(all P<0.05),whereas only IPI had a significant effect on OS(P<0.001).The incidence of leukopenia and grade 3-4 leukopenia in the separated R-CHOP group was significantly lower than that in the full-dose R-CHOP group(P=0.007,P=0.012),but there was no significant difference with the reduced R-CHOP-like group in this regard.Conclusions:In older patients with newly diagnosed DLBCL,separated R-CHOP showed good efficacy both in the short and long terms and had acceptable safety and tolerability profiles.

Objective:To improve the therapeutic regimen for mantle cell lymphoma,we investigated the efficacy and safety of adding a BTK inhibitor to a regimen including rituximab,bendamustine,cytarabine,and prednisone to treat patients with mantle cell lymphoma(MCL).Methods:Twenty-six patients newly diagnosed with MCL who were admitted to The First Affiliated Hospital of Zhengzhou University from March 2021 to November 2023 were treated with a regimen of rituximab,bendamustine,cytarabine and prednisone combined with a BTK inhibitor,and the efficacy and adverse effects of this regiment were retrospectively analyzed.Results:The median age of the 26 newly dia-gnosed MCL patients was 59(41-72)years.The cohort included 22 males and 4 females,and the median follow-up time was 12(3-28)months.The overall response rate(ORR)was 92.3%and the complete response rate(CRR)was 88.5%.Median progression-free survival(PFS)and median overall survival(OS)endpoints were not achieved,with a 1-year PFS rate of 81.25%and a 1-year OS rate of 92.3%.A bet-ter PFS was achieved in the low mantle cell lymphoma International Prognostic Index(MIPI)score(0-3 points)group than in the high MIPI score(4-11 points)group(P=0.020).PFS was better in the group without B symptoms than in the group with B symptoms(P=0.002).PFS was better in the classical group than in the pleomorphic-blastoid subtype group(P=0.009).The main adverse effects were lymphopenia and thrombocytopenia.No treatment-related serious adverse events were observed during the follow-up period.Conclusions:The regimen of rituximab,bendamustine,cytarabine,and prednisone in combination with BTK inhibitors is safe and effective for the treatment of newly dia-gnosed patients with MCL.

ObjectiveTo observe the possible mechanism of Dihuang Yinzi Decoction （地黄饮子） for improving cognitive dysfunction in Alzheimer's disease （AD） from the perspective of retina. MethodsForty-five APP/PS1 mice （AD model mice） were randomly divided into model group， Dihuang Yinzi Decoction group， and memantine group， with 15 mice in each group， while 15 wild-type C57BL/6J mice from the same litter were used as blank group. Mice in Dihuang Yinzi Decoction group were given Dihuang Yinzi Decoction 30.03 g/（kg·d） by gavage， mice in the memantine group were given memantine hydrochloride 6.1 mg/（kg·d） by gavage， and mice in the blank group and the model group were given normal saline 2 ml/（kg·d） by gavage for 4 consecutive weeks. Fasting blood glucose was measured weekly. After 4 weeks of intervention， oral glucose tolerance test （OGTT） and insulin tolerance test （ITT） were performed； Morris water maze was used to detect the changes in spatial memory ability of mice； glucose oxidase method was used to detect retinal glucose content of mice； enzyme-linked immunosorbent assay （ELISA） was used to detect serum and retinal insulin content of mice， and Homeostatic Model Assessment of insulin resistance （HOMA-IR） was calculated. Hematoxylin-eosin （HE） staining was performed to observe the histopathological changes in the retina， and the retinal thickness and ganglion cell number were counted； protein immunoblotting was performed to detect the retinal pathway-associated proteins ［insulin receptor substrate 1 （IRS1）， phosphorylated insulin receptor substrate 1 （pIRS1）， phosphatidylinositol-3-hydroxykinase （PI3K）， protein kinase B （Akt1）， phosphorylated protein kinase B （pAkt1）］ expression； retinal glucose transporter protein 4 （GLUT4） expression was detected by immunohistochemistry. ResultsCompared with the blank group， fasting blood glucose of mice in the model group at weeks 1， 2， 3， and 4， blood glucose and area under the curve （AUC） at different time point of OGTT and ITT test， fasting serum insulin， and HOMA-IR increased （P＜0.05， P＜0.01）； in the Morris water maze experiment， the escape latency increased from day 3 to day 5， and the number of crossing platforms， the percentage of target quadrant distance， and the percentage of target quadrant time decreased （P＜0.05， P＜0.01）； the outer nuclear layer of the retina became sparse， thinner， and the number of ganglion cells decreases （P＜0.01）； the expression level of retinal glucose increased， while the expression levels of insulin， pIRS1/IRS1， PI3K/β-Actin， pAkt1/Akt1， and GLUT4 proteins decreased （P＜0.01）. Compared with the model group， fasting blood glucose at week 4， blood glucose at each time point of the OGTT and ITT tests AUC decreased （P＜0.05， P＜0.01）， and fasting serum insulin and HOMA-IR decreased （P＜0.05） in Dihuang Yinzi Decoction group； In the Morris water maze test， the escape latency shortened on day 4 and day 5， number of platform crossings， target quadrant distance as a proportion of total distance， and target quadrant movement time as a proportion of total time decreased （P＜0.05， P＜0.01）； retinal pathological changes alleviated， and retinal thickness and ganglion cell number increased （P＜0.01）； retinal glucose content decreased， and retinal pIRS1/IRS1， PI3K/β-Actin， and GLUT4 protein expression elevated （P＜0.05 or P＜0.01）. ConclusionsDihuang Yinzi Decoction can improve cognitive dysfunction of Alzheimer's disease， which may be related to regulating retinal insulin content and insulin signaling pathway.

OBJECTIVETo investigate the role of holoenzyme containing Protein Phosphatase 2A B56β in regulating CdCl2 induced cytotoxicity.

METHODCdCl2-induced cytotoxicity in normal human cell line L-02, AFB1-transformed hepatic cell line L-02 RT-AFB1 and tumor cell line Bel7402 was measured by modified MTT assay. Stable cell lines L-02 SHAKT, L-02 SHB56β, L-02 RT-AFB1-B56β and Bel7402-B56β were generated by infecting L-02 cells or Bel7402 cells with retroviral vectors encoding lentiviral AKT shRNA, lentiviral B56β shRNA and B56β. The relative cell viability was measured in normal human cell line AFB1-transformed hepatic cell line and tumor cell line when treated by CdCl2 (0, 20, 40, 80, 160 µmol/L). After treated by wortmannin (2.5, 5.0 µmol/L) combined with 40 µmol/L CdCl2, Western blot was applied to measure the expression of associated protein in L-02.Western blot was applied to measure the expression of B56β, MT (metallothionein), AKT, and p-AKT in these cell lines treated by CdCl2.

RESULTSThe levels of MT were 0.12 ± 0.02, 0.06 ± 0.06 in L-02 RT-AFB1 and Bel7402, which were lower than L02 (0.92 ± 0.14) (F = 1 148.16, P < 0.001) when treated by 40 µmol/L CdCl2. When treated by 40 µmol/L CdCl2, the expression of p-AKT in L-02 SHAKT-1 and L-02 SHAKT-2 were 0.08 ± 0.02, 0.08 ± 0.05, which levels were lower than L-02 SHGFP (0.18 ± 0.15) (F = 724.70, P < 0.001); and the expression of MT were both 0.62 ± 0.16 in L-02 SHAKT-1 and L-02 SHAKT-2, which levels were higher than L-02 SHGFP (0.22 ± 0.14) (F = 94.73, P < 0.001). After treated by wortmannin (2.5, 5.0 µmol/L) combined with 40 µmol/L CdCl2, the expression of p-AKT in L-02 were 0.28 ± 0.07, 0.15 ± 0.11, which levels were lower than wortmannin untreated cells (0.52 ± 0.11) (F = 578.57, P < 0.001); and the expreesion of MT were 1.62 ± 0.80, 1.08 ± 0.15, which levels were higher than wortmannin untreated cells (0.69 ± 0.18) (F = 12.34, P < 0.001). When treated by 40 µmol/L CdCl2, the levels of p-AKT in L-02 SHB56β-1 and L-02 SHB56β-2 were 0.57 ± 0.13, 0.59 ± 0.02, which were higher than L-02 SHGFP (0.32 ± 0.02) (F = 87.16, P < 0.001); and the levels of MT were 0.35 ± 0.07, 0.20 ± 0.03 in L-02 SHB56β-1 and L-02 SHB56β-2, which were lower than L-02 SHGFP (1.51 ± 0.13) (F = 2 457.10, P < 0.001). After treated by 40 µmol/L CdCl2, the expression of p-AKT in L-02 RT-AFB1-B56β and Bel7402-B56β were 0.10 ± 0.11, 0.09 ± 0.01, which were lower than L-02 RT-AFB1 (0.36 ± 0.01) and Bel7402 (0.43 ± 0.11) (F = 877.62, P < 0.001); and the levels of MT were 0.92 ± 0.13, 0.95 ± 0.08 in L-02 RT-AFB1-B56β and Bel7402-B56β,which were higher than L-02 RT-AFB1 (0.44 ± 0.12) and Bel7402 (0.77 ± 0.06) (F = 51.97, P < 0.001).

CONCLUSIONProtein phosphatase 2A complexes containing B56β participated in the regulation of MT expression through direct dephosphorylation of AKT, finally affected the cytotoxicity responding to CdCl2. Our study revealed a key signaling pathways of PP2A involved in heavy metals induced cytotoxicity.

Cadmium Chloride ; Cell Line ; Cell Line, Tumor ; Cell Survival ; Holoenzymes ; Humans ; Liver ; Metallothionein ; Metals, Heavy ; Protein Phosphatase 2 ; Signal Transduction