Objective:To study the effects of Zuogui Pills on rats with kidney-yin deficiency syndrome of premature ovarian insufficiency.Methods:Totally 40 SD female unmated rats were randomly divided into blank group, model group, Zuogui Pills group and Bujiale group, with 10 rats in each group. Except for blank group, rats in other groups were subcutaneously injected with ZP3 and gavaged with levothyroxine sodium to induce kidney-yin deficiency syndrome model of premature ovarian insufficiency. At the same time of modeling, Zuogui Pills group and Bujiale group received corresponding drugs for gavage, and the other groups received corresponding solvent for gavage, once a day, for consecutive 21 days. On day 0, 7, 14 and 21, ear temperature and body weight of rats were measured, and the ovarian index, uterus index and thyroid index were calculated. Serum levels of adenosine cyclic phosphate (cAMP), cyclic guanosine phosphate (cGMP), Cortisol, follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E 2) were detected by ELISA. Pathological changes of ovary was observed with HE staining. Results:On day 14 and 21, compared with model group, the body weight of rats in Zuogui Pills group increased ( P<0.05), and the ear temperature decreased ( P<0.05); compared with model group, the ovarian index, uterine index and thyroid index of rats in Zuogui Pills group decreased ( P<0.05), the levels of serum cAMP/cGMP, cortisol, FSH and LH decreased ( P<0.05), and the level of E 2 increased ( P<0.05). Conclusion:Zuogui Pills have certain improvement effect on rats with kidney-yin deficiency syndrome induced by levothyroxine sodium tablets combined with ZP3.

Objective:To study the performance of immune reconstitution in patients with chimeric antigen receptor (CAR)-T cell immunotherapy bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:A total of 61 patients with acute B lymphocytic leukemia (B-ALL) who received CAR-T cell bridging allo-HSCT in Beijing Lu Daopei Hospital from August 2018 to December 2021 were enrolled, and the clinical medical records of the above patients were retrospectively analyzed. The average age was 14 (7, 30) years old, including 39 males and 22 females. 32 patients were treated with CAR-T cell immunotherapy(CAR-T Group) and 29 didn't with CAR-T cell immunotherapy(non-CAR-T group). The follow-up period was 561 (235,784) days. Multicolor flow cytometry was used to detect the peripheral blood lymphocyte subsets, i.e. total lymphocytes, T lymphocytes, helper T cells, cytotoxic T cells, B lymphocytes, NK cells, and Treg cell counts before transplantation and 1, 2, 3, 6, 8, 10, and 12 months after transplantation, to evaluate the immune reconstitution performance post allo-HSCT.Results:Serum globulin before transplantation: The IgA level in the CAR-T group was 0.18 (0.06, 0.49) g/L, which was lower than that of 1.03 (0.63, 1.56) g/L in the non-CAR-T group ( U=103.5, P<0.001). The IgG level in the CAR-T group was 5.54 (4.04, 7.09) g/L, lower than that of 6.78 (5.27, 9.26) g/L in the non-CAR-T group, ( U=1 298.5, P=0.017), and the IgM level in the CAR-T group was 0.18 (0.05, 0.30) g/L, lower than that of 0.40 (0.26, 0.71) g/L in the non-CAR-T group ( U=166.0, P<0.001). In the CAR-T group before transplantation, the absolute count of total lymphocyte in peripheral blood was 833.00 (335.00, 1 727.50) /μl, lower than that of 1 052.00 (545.75, 1 812.50) /μl in the non-CAR-T group ( U=404.0, P<0.001). The absolute count of T lymphocyte in the CAR-T group before transplantation was 686.00 (233.00, 1 307.00)/μl, lower than that of 860.00 (391.00, 1 419.75) /μl in the non-CAR-T group ( U=406.0, P<0.001). The absolute count of helper T lymphocytes in the CAR-T group was 146.00 (40.50, 327.50) /μl, lower than that of 162.50 (66.00, 384.75) /μl in the non-CAR-T group ( U=494.0, P=0.002). The absolute count of cytotoxic T lymphocytes in the CAR-T group was 343.00 (56.50, 924.00) /μl, lower than that of 478.00 (143.50, 992.25) /μl in the non-CAR-T group ( U=483.5, P=0.001). The absolute count of B lymphocytes in CAR-T group was 22.00 (6.00, 186.00) /μl, lower than that of 33.00 (8.00, 220.00) /μl in the non-CAR-T group ( U=498.0, P=0.002). And when two groups of patients were monitored after transplantation, there was no statistical difference in absolute cell counts of each immune cell subpopulation( P>0.05). Comparing the clinical features of the two groups, the pre-transplant history of the CAR-T group was 981.00 (368.50, 1 514.75) d, longer than that of 323.00 (167.50, 450.50) d in the non-CAR-T group ( U=263.0, P=0.004). The dose of rabbit anti-human thymic immunoglobulin (ATG) in the pretreatment protocol of patients in the CAR-T group was 5.00 (5.00, 7.50) mg/Kg, lower than that of 7.00 (5.00, 7.50) mg/kg in the non-CAR-T group ( U=288.5, P=0.018). The infusion dose of CD34 +cells in the CAR-T group was 5.91 (4.23, 6.02) ×10 6/kg, higher than that of 4.51 (4.00, 5.93)×10 6/kg in the non-CAR-T group ( U=291.0, P=0.012). The duration of the application of cyclosporine after transplantation in the CAR-T group was 167.00 (119.25, 299.50) d, which was shorter than that of 197.00 (102.50, 450.50) d in the non-CAR-T group ( U=421.0, P=0.001). Conclusions:For patients in CAR-T group with low immune function before transplantation, it may be possible to make them comparable to non-CAR-T group in immune reconstitution state by reducing the dose of pretreatment ATG, increasing the counts of CD34 + cells infusion in the graft, and discontinuing cyclosporine as soon as possible after transplantation.

Objective:To investigate the effects of different concentrations of hydrogen peroxide (H 2O 2) on the expression of histone deacetylase-2(HDAC-2) and mitogen activated protein kinases phosphatases-1(MKP-1) in A549 cells. Methods:The experimental models of A549 cells were established by intervening of different concentrations of H 2O 2(0, 100, 200, 400, 600, 800 μmol/L)for 2 h, 4 h, 8 h, 12 h and 24 h, respectively.The survival rate of A549 cells was detected by thiazolyl blue(MTT) assay, and the apoptosis of A549 cells was tested by lactic dehydrogenase(LDH) assay.The expression of HDAC-2 and MKP-1 protein was detected by Western blot. Results:(1)MTT results showed that compared with the control group (H 2O 2 was 0 μmol/L), the survival rate of A549 cells was decreased with the increase of H 2O 2 concentration at 2 h, 4 h, 8 h, 12 h, 24 h and 48 h. (2)LDH results showed that, compared with the control group, the inhibition rate of A549 cells increased with the increase of H 2O 2 concentration at 2 h, 4 h, 8 h, 12 h, 24 h and 48 h. (3)The expression of HDAC-2 protein decreased ( F=14.588, P<0.01), and the expression of MKP-1 protein increased ( F=64.297, P<0.01) in a concentration-dependent manner with the increase of H 2O 2 concentration at 12 h after H 2O 2 treatment. Conclusions:H 2O 2 participates in oxidative stress injury of lung cells by regulating the expression of HDAC-2 and MKP-1.

Objective:To explore the effectiveness of autologous fat grafting in the treatment of undesirable skin expansion.Methods:Patients' data were reviewed from 2011 to 2016, including the expanded regions with early signs of skin complications in face and neck. The effects of fat grafting group and control group were evaluated by follow-up records of expansion volume, skin thickness, skin texture and local capillary reaction.Results:Fat grafting could increase the thickness of expanded skin. It also improved the texture of expanded skin, with 0.83± 0.71 points before treatment and 1.30±0.66 points after treatment ( P=0.04). The local capillary reaction was also improved from 1.06±0.54 points before treatment and 1.45±0.51 points after treatment ( P=0.03). The expansion in the fat grafting group was 2.21±0.57 times before treatment and 2.94±0.83 times after treatment. In the control group, the expansion was 2.19 times when it showed early signs, and no obvious changes were observed during the follow-up period. Conclusions:Autologous fat grafting can effectively treat complications of skin expansion, prolong expansion process and promote tissue regeneration.

Bronchopulmonary dysplasia (BPD) is a common chronic respiratory complication in preterm infants without fully understand the mechanism or effective treatment, which could significantly affect the survival rate and prognosis of these infants. Studies have confirmed that epigenetic mechanisms, including histone modification, non-coding RNA and DNA methylation may play an essential role in the onset and development of BPD. And most related epigenetic changes are reversible, which might serve as a potential target for BPD treatment. Therefore, further studies on epigenetics will shed light on a better understanding of the pathogenesis, prevention, and treatment of BPD.

Objective:The purpose of this study is to establish the trajectory of targeted grafting for facial fat compartment based on anatomical research, and then bring it to clinical practice.Methods:The boundary of fat compartment and the relationship of adjacent vessel and nerve were clarified through autopsy. The recommended injection points and trajectory for targeted fat grafting were established on the anatomical findings. Retrospective clinical data of facial rejuvenation of 46 patients through targeted fat grafting were collected from June 2017 to June 2019 in Shanghai Ninth People’s Hospital. The result of 3D scanning were analyzed to evaluate the survival rate of fat grafts.Results:There were subcutaneous superficial fat compartments in the frontal region, and there were both deep and superficial fat compartments in the temporal and middle face. According to the anatomical characteristics, a targeted fat grafting technique was established with the frontal hairline and the oral commissure corner mucosa as the entry points. In the clinical study, 46 patients were evaluated by 3D scanning 6 months after the last fat grafting. The amount of fat grafts in the temporal region was (17.84±8.47) ml and (11.2±2.44) ml was left after operation, and the survival rate was 63%. The amount of fat grafts in mid-face was (26.81±10.36) ml and (16.09±4.48) ml was left after operation, and the survival rate was 60%. Overall satisfaction rate of patients was 93% (43/46).Conclusions:Compartment-based targeted fat grafting is an accurate injection method, which meets the requirement of physiological augmentation. The trajectory of targeted fat grafting will further improve the efficacy and safety of injection.

Objective:The purpose of this study is to establish the trajectory of targeted grafting for facial fat compartment based on anatomical research, and then bring it to clinical practice.Methods:The boundary of fat compartment and the relationship of adjacent vessel and nerve were clarified through autopsy. The recommended injection points and trajectory for targeted fat grafting were established on the anatomical findings. Retrospective clinical data of facial rejuvenation of 46 patients through targeted fat grafting were collected from June 2017 to June 2019 in Shanghai Ninth People’s Hospital. The result of 3D scanning were analyzed to evaluate the survival rate of fat grafts.Results:There were subcutaneous superficial fat compartments in the frontal region, and there were both deep and superficial fat compartments in the temporal and middle face. According to the anatomical characteristics, a targeted fat grafting technique was established with the frontal hairline and the oral commissure corner mucosa as the entry points. In the clinical study, 46 patients were evaluated by 3D scanning 6 months after the last fat grafting. The amount of fat grafts in the temporal region was (17.84±8.47) ml and (11.2±2.44) ml was left after operation, and the survival rate was 63%. The amount of fat grafts in mid-face was (26.81±10.36) ml and (16.09±4.48) ml was left after operation, and the survival rate was 60%. Overall satisfaction rate of patients was 93% (43/46).Conclusions:Compartment-based targeted fat grafting is an accurate injection method, which meets the requirement of physiological augmentation. The trajectory of targeted fat grafting will further improve the efficacy and safety of injection.

In order to enrich the library of domestic research about new red fluorescent marker in lactic acid bacteria (LAB), we described a new fusion expression system in Lactobacillus plantarum WCFS1 based on the pSIP vector. This system contained red fluorescent protein mCherry as a marker and bile salt hydrolase gene (bsh) as a reporter gene. Moreover, in this study, four different promoters (PsppA, PldhL, P32 and PslpA) were used to regulate the expression of the fusion protein mCherry-BSH, completing the inducible and constitutive expression in lactic acid bacteria. The recombinant protein mCherry-BSH presented activity of red fluorescence and bile salt hydrolase (BSH). The successful construction of the fusion expression system in LAB using a red fluorescent protein mCherry provides favorable conditions for the distribution, intestinal colonization and survival rate of lactic acid bacteria, so as to reveal the function mechanism of its probiotic characteristics; and the system also could lay the foundation for researches on protein expression, cellular localization and properties identification of active protein in lactic acid bacteria.
Lactobacillus plantarum ; Luminescent Proteins ; Probiotics

Objective To investigate the effect of NS-398 combined with docosahexaenoic acid (DHA) on apoptosis of cholangiocarcinoma QBC939 cells and its mechanism.Methods In vitro,cultured cholangiocarcinoma QBC939 cells were exposed to 0,25,50,100,150 and 200 μmol/L NS-398 with 0,15,30,45,60 and 75 μg/ml DHA respectively.The absorbance of the QBC939 cells were measured by CCK8 and its growth inhibition ratio were calculated.Flow cytometry was applied to detect cell apoptosis.The level of β-catenin and c-myc mRNA and protein were measured by real-time PCR,Western blot and enzyme-linked immunoabsordent assay respectively.Results Exposure to NS-398 combined with DHA suppressed the growth of QBC939 cells.When NS-398 was at 100 μmol/L and DHA at 45 pμg/ml,the relative growth inhibition rate of QBC939 cells was 90％ (F =5.85,P ＜ 0.05).NS-398 combined with DHA promoted QBC939 cells apoptosis at the early stage (F =8.16,P ＜ 0.01).Real-time PCR could detect low β-catenin and c-myc expression in QBC939 cells disposed by NS-398 combined with DHA (F =7.61,P ＜0.01),(F =7.92,P ＜0.01).NS-398 combined with DHA decreased β-catenin (F =7.75,P ＜ 0.01),(F=8.17,P＜0.01) and c-myc (F=8.76,P＜0.01),(F=8.12,P＜0.01) protein expression in QBC939 cells.Conclusion NS-398 combined with DHA promoted apoptosis and inhibit proliferation of cholangiocarcinoma cells QBC939 in vitro possibly through downregulated mRNA and protein expression of β-catenin and c-myc.