Objective: To investigate the effect of all trans retinoid acid (ATRA) on bone cell proliferation and differentiation. Methods: Newborn rat calvarial osteoblastic cells were isolated and the metabolism of the osteoblastic cells were determined by MTT, Goldens method and immunocytohistologic method. Results: 10 -5 , 10 -6 , 10 -7 , 10 -8 , 10 -9 mol/L ATRA could increase osteoblastic cells proliferation after 72 h culture; 10 -5 , 10 -6 , 10 -7 mol/L ATRA could increase ALP activity. The expression of cyclin D 1 was decreased. Conclusion: ATRA stimulates the proliferation and differentiation of rat calvarial osteoblastic cells in vitro. Cell cycle relative proteins may play an important role in control of cell proliferation and differentiation induced by retinoic acid and derivatives.

Objective: Adopting quantified index to observe and analyze the effect of acute and chronic pain through double-wavelength diode laser. Methods: Using diode laser with the wavelength of 650nm / 810nm to irradiate the special tissues or aching points of patient, selecting different frequency, intensity and time to make treatment. Results: VAS scoring: P<0.05, the total efficient rate is 96%. Conclusions: Treatment with double-wavelength diode laser to various acute and chronic pain is just a definitely good curative effect, and it can release the effects of patient sleeping, daily life, communication with people and life interests caused by pain, and finally enhancing the quality of life.

The knock out vector pHK was constructed with E coli vector pET 28a and shuttle vector pHY300PL, by using denatured DNA and homologous recombination technique, the kanamycin resistance gene ( Kan r) from integrated alkaline protease gene engineering strain BP071 was knocked out successfully, and the 11 positive clones were obtained The yield of the best positive clone BP0715 was stable as same as BP071 The methods provided the good experience for the industrial microbiology research, and it was foundation for studying on the safety of genetically modified organisms

Wuxistatin, a novel and potent statin, is converted from lovastatin by Amycolatopsis sp. CGMCC1149. In the bioconversion, lovastatin is firstly hydroxylated by a hydroxylase. To obtain the critical hydroxylase, a novel hydroxylase gene was isolated from Amycolatopsis sp. CGMCC1149 by Degenerate PCR and Self-Formed Adaptor PCR and expressed in Escherichia coli. BLAST sequence analysis revealed that the gene belonged to cytochrome P450 gene superfamily and could encode a 403-amino-acid protein with a molecular weight of 44.8 kDa. The secondary structure prediction result showed that this protein contained many typical functional regions of P450, such as oxygen binding site, ion-pair region and heme binding region. Meanwhile, a catalytic function verification system was constructed by NADH, ferredoxin and ferredoxin reductase which could catalyze lovastatin hydroxylation into the target product. These would be helpful for further studies in large-scale production of wuxistatin.
Actinomycetales ; enzymology ; genetics ; Amino Acid Sequence ; Butyrates ; metabolism ; Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Hydroxylation ; Industrial Microbiology ; Lovastatin ; metabolism ; Molecular Sequence Data

Objective To explore the contribution of ultrasonic examination and localization in early breast cancer screening of chinese women with dense breast.Methods From February,2002 to April,2006,the breasts of 5 000 women were examined using ultrasonic examination,and in about 4 000 women a mass was detected,which included 142 cases of breast cancer proved by pathology with diameter ≤2cm occurred in patients with dense breast.In these parients,ultrasonic visualization and molybdenum target mammographic were performed.Results In these with breast cancer and dense breast,there were 47 cases(33.10%) with microcalcification detected by ultrasonography,the sensitivity rate was 74.60% and the accuracy rate was 88.73%.Only 38(26.76%) cases were detected by mammography,the sensitivity rate was(60.32)% and the accuracy was 82.39%.About 44.37% breast cancer with dense brast display as the(microcalcification).There were 110 cases(77.46%) of the small breast cancer detected by ultrasonography,the sensitivity was 88.71% and the accuracy rate was 90.14%.There were 100 cases(70.42%)(detected) by mammography,the sensitivity rate was 80.65% and the accuracy rate was 83.10%.Also,in 12 patients an unpalpation breast lesion was correctly excised by the use of ultrasonic localization.(Conclusions)(1)For the small breast cancer in dense breast the sensitivity and the accuracy of ultrasonography(surpassed) those of mammography.(2)The ultrasonic examination is an effective way for early breast cancer screening in young women with dense breast and flat breast.Ultrasonography can improve the rate of dignosis in early breast cancer,and also improve the breast-conserving surgical rate.Ultrasonography is worth of widespread use spreading.

Objective To investigate the characteristics and failure risk factors of sequential high-flow nasal cannula oxygen therapy (HFNC) after weaning from invasive ventilation. Methods The patients who received sequential HFNC after weaning from invasive ventilation admitted to surgical intensive care unit (ICU) of Peking University People's Hospital from June 1st 2016 to May 31st 2018 were retrospectively analyzed. Clinical variables, respiratory therapy parameters, respiratory variables, cardiac variables and outcomes were reviewed and analyzed. Treatment characteristics of HFNC after weaning was analyzed. Patients were divided into HFNC success group and HFNC failure group according to the failure of HFNC, and the differences between the two groups were compared. The independent risk factors of HFNC treatment failure were analyzed by Logistic regression analysis. The value of predictive treatment failure of risk factors and regression models were analyzed by receiver operating characteristic (ROC) curve. Results A total of 99 patients were included, 61 men, and the median age was 67.0 (57.0, 76.0) years old. The medianinitial HFNC flow was 50 (50, 60) L/min, and inspired oxygen concentration (FiO2) was 0.50 (0.40, 0.60). Eighteen patients experienced HFNC failure (18.2%). Compared with the HFNC success group, the sequential organ failure assessment (SOFA) score in the HFNC failure group was higher [4 (3, 5) vs. 2 (1, 3), P < 0.01], B type natriuretic peptide (BNP) before HFNC therapy were significant higher [ng/L: 647.2 (399.2, 1 331.3) vs. 127.2 (55.2, 369.5), P < 0.01], and respiratory frequency (RR) and heart rate (HR) were significant faster, mean arterial pressure (MAP) was significant higher, oxygen index (PaO2/FiO2) was significant lower after 30 minutes HFNC treatment [RR (times/min): 26 (22, 28) vs. 19 (17, 21), HR (bpm): 105 (97, 107) vs. 85 (77, 90), MAP (mmHg, 1 mmHg = 0.133 kPa): 104.3 (101.7, 110.7) vs. 92.3 (88.3, 97.7), PaO2/FiO2 (mmHg): 207.3 (185.8, 402.8) vs. 320.2 (226.2, 361.5), all P < 0.05]. It was shown by multiple Logistic regression analysis that the SOFA score [odds ratio (OR) = 2.818, P = 0.022, β = 1.036], BNP before HFNC treatment (OR = 1.002, P = 0.033, β = 0.002) and HR after HFNC treatment 30 minutes (OR = 1.140, P = 0.032, β = 0.131) were independent risk factors for HFNC treatment failure. It was shown by ROC curve that the area under the ROC curve (AUC) for the prediction of HFNC failure was 0.840, 0.859, 0.860 and 0.962 for SOFA, BNP before HFNC treatment, HR after HFNC treatment 30 minutes, and regression model, all had good forecast values (all P < 0.01). Conclusions HFNC is one of the commonly used oxygen therapy methods in the ICU, but not all patients who are treated as a sequential therapy after invasive mechanical ventilation weaning can benefit from it. SOFA score, BNP before HFNC treatment and HR after 30 minutes HFNC treatment were independent risk factors of HFNC failure. Each independent risk factor and regression model can predict the success of HFNC treatment.

Candida glycerinogenes WL2002-5 (C.g) is an important industrial strain for glycerol production. To further improve glycerol production, we reconstructed a binary vector pCAM3300-zeocin-CgGPD1, introduced it to Agrobacterium tumefaciens LBA4404 by electroporation, and then transformed the T-DNA harboring the CgGPD1 to Candida glycerinogenes by Agrobacterium tumefaciens-mediated transformation (ATMT). After 96 h fermentation with glucose as the substrate, we screened a transformant named C.g-G8 with high glycerol production. Compared with the wild strain, the glucose consumption rate of C.g-G8 and the glycerol production were 12.97% and 18.06% higher, respectively. During the fermentation, the activity of glycerol-3-phosphate dehydrogenase of C.g-G8 was 27.55% higher than that of the wild strain. The recombinant Candida glycerinogenes with high glycerol production was successful constructed by ATMT method.
Agrobacterium tumefaciens ; enzymology ; genetics ; Candida ; genetics ; metabolism ; Electroporation ; Fermentation ; Glycerol ; analysis ; metabolism ; Glycerolphosphate Dehydrogenase ; genetics ; Recombination, Genetic ; Transformation, Genetic

Objective:To investigate the early diagnostic value of red blood cell distribution width(RDW)in elderly patients with acute ischemic stroke(AIS), and to explore whether RDW can be used as an independent predictor of AIS.Methods:This retrospective study included elderly AIS patients who were hospitalized within the first 24 hours after the onset of symptoms from November 2018 to October 2019 as the AIS group, and elderly patients with various conditions that masqueraded as a stroke who were admitted to hospital during the same period were included as the control group.The demographic data, vascular risk factors, and baseline RDW levels were recorded, and the receiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of RDW for AIS.Results:The median level of RDW in the AIS group was significantly higher than that in the control group(13.0% vs.12.4%, P<0.001). The area under the curve of RDW was 0.758(95% CI: 0.685-0.823). At the cut-off value was 12.7%, and its sensitivity and specificity were 73.2% and 75.3%, respectively.Multivariate Logistric regression analysis showed that after adjusting for covariates, the risk of AIS is 5.342-fold for each 1% increase in RDW( OR: 5.342, 95% CI: 2.520-11.324, P<0.001). Conclusions:The increased level of RDW is related to the risk of AIS, and it is an independent predictor of AIS.Thus, it may be used as a preliminary screening index for the diagnosis of AIS.

ObjectiveTo explore the possible mechanism of Shipi Xiaoji Beverage （实脾消积饮） in treating primary liver cancer from the perspective of mitochondrial dynamics imbalance and ferroptosis. MethodsThe Shipi Xiaoji Beverage-containing serum was prepared， and liver cancer HepG2 cells were cultured in vitro. The blank group， control group， cisplatin group （10 µg/ml）， and 5%， 10%， and 15% medicated serum groups were set， with 4 duplicate holes in each group. After adding the corresponding medicinals to each group， the cell survival rate was calculated using the CCK-8 test after 24-hour culture at 37 ℃ with 5% CO₂. Using transwell invasion assay， the number of cells that reached the upper chamber membrane through penetrating the Matrigel was calculated after culturing for 48 hours. The scratch migration rate was calculated after incubation for 24 h and 48 h in the scratch experiment， and the concentration of the medicated serum was screened for subsequent experiments. The control group， 10% medicated serum group （the screened concentration） and 10% medicated serum+Mdivi-1 group were set up. After culturing for 24 hours， the average fluorescence intensity of mitochondria and reactive oxygen species （ROS）， adenosine triphosphate （ATP） content and level were measured， and the protein expression of cytoplasmic and mitochondrial dynein-related protein 1 （Drp1） and mitochondrial phosphorylated dynamin-related protein 1 （p-Drp1） was detected. The control group， 10% medicated serum group， ferroptosis activator group （Erastin， 10 μmol/L） and 10% medicated serum+ferroptosis inhibitor group （fer-1，10 μmol/L） were set up， and after adding the corresponding medicinals to each group and culture for 24 hours， the intracellular glutathione （GSH）， malondialdehyde （MDA） content and ferrous ion level were detected， as well as the protein expression of solute carrier family 7 member 11 （SLC7A11） and glutathione peroxidase 4 （GPX4） in cells. ResultsCompared to those in the control group， the cell survival rate， invasion number， 24 h and 48 h scratch migration rate in all medicated serum groups and cisplatin group significantly decreased （P＜0.01）； as the concentration of medicated serum increased， the cell survival rate， invasion number and 48h scratch migration rate of each medicated serum group gradually decreased （P＜0.01）， and finally， 10% of Shipi Xiaoji Beverage-containing serum was selected for subsequent experimental concentration. Compared to the control group， the 10% medicated serum group had decreased average fluorescence intensity of mitochondria and ATP content and increased average fluorescence intensity of ROS （P＜0.05）； compared to the 10% medicated serum group， the 10% medicated serum+Mdivi-1 group had higher average fluorescence intensity of mitochondria and ATP content and lower average fluorescence intensity of ROS （P＜0.05 or P＜0.01）. Compared to those in the control group， GSH content and protein expression of SLC7A11 and GPX4 in other groups significantly decreased， while MDA content and ferrous ion level increased （P＜0.05 or P＜0.01）； compared to the ferroptosis activator group， the 10% medicated serum group and the 10% medicated serum+ferroptosis inhibitor group had decreased MDA content and ferrous ion level， and increased protein expression of SLC7A11 and GPX4 （P＜0.05 or P＜0.01）； the 10% medicated serum+ferroptosis inhibitor group had higher GSH level than the 10% medicated serum group （P＜0.01）. ConclusionShipi Xiaoji Beverage may inhibit the proliferation， infestation， migration of HepG2 cells by causing mitochondrial dynamics imbalance and inducing ferroptosis， thereby playing against liver cancer.

Objective:Exploring the efficacy and safety of bridging blinatumomab (BiTE) in combination with chimeric antigen receptor T (CAR-T) cell therapy for the treatment of adult patients with acute B-cell lymphoblastic leukemia (B-ALL) .Methods:Clinical data from 36 adult B-ALL patients treated at the First Affiliated Hospital of Suzhou University from August 2018 to May 2023 were retrospectively analyzed. A total of 36 cases were included: 18 men and 18 women. The median age was 43.5 years (21-72 years). Moreover, 21 cases of Philadelphia chromosome-positive acute lymphoblastic leukemia were reported, and 16 of these cases were relapsed or refractory. Eighteen patients underwent blinatumomab bridging followed by CAR-T cell therapy, and 18 patients received CAR-T cell therapy. This study analyzed the efficacy and safety of treatment in two groups of patients.Results:In the BiTE bridge-to-CAR-T group, 16 patients achieved complete remission (CR) after BiTE immunotherapy, with a CR rate of 88.9%. One month after bridging CAR-T therapy, bone marrow examination showed a CR rate of 100.0%, and the minimal residual disease (MRD) negativity rate was higher than the nonbridging therapy group (94.4% vs. 61.1%, Fisher, P=0.041). The incidence of cytokine release syndrome and other adverse reactions in the BiTE bridge-to-CAR-T group was lower than that in the nonbridging therapy group (11.1% vs. 50.0%, Fisher, P=0.027). The follow-up reveals that 13 patients continued to maintain MRD negativity, and five patients experienced relapse 8.40 months (2.57-10.20 months) after treatment. Two of five patients with relapse achieved CR after receiving the second CAR-T cell therapy. In the nonbridging therapy group, 10 patients maintained continuous MRD negativity, 7 experienced relapse, and 6 died. The 1 year overall survival rate in the BiTE bridge-to-CAR-T group was higher than that in the nonbridging therapy group, with a statistically significant difference at the 0.1 level (88.9%±10.5% vs. 66.7%±10.9%, P=0.091) . Conclusion:BiTE bridging CAR-T cell therapy demonstrates excellent efficacy in adult B-ALL treatment, with a low recent recurrence rate and ongoing assessment of long-term efficacy during follow-up.