Parkinson's disease is a progressive neurodegenerative disorder in central nervous system caused by the loss of midbrain dopaminergic neurons. At present, dopaminergic neurons differentiated both in vivo and in vitro from neural stem cells act as an important cell source for the cell-replacement therapy of Parkinson's disease. This paper reviewed the major molecular mechanism, signaling pathway and important environment factors involved in differentiation of dopaminergic neurons from neural stem cells, based on the research findings in recent 10 years.

Objective:To investigate the effect of steroid hormones on distribution of uterine natural killer(uNK)cells in the mouse uterus.Methods:A unique uNK cell marker, Dolichos biflorus agglutinin(DBA)lectin was used to localize uNK cells in the ovariectomized and ovarian steroid hormone-treated mouse uterus by immunohistochemical staining.Results: After estrogen(E_2)was administered in the ovariectomized mice, uNK cells were distributed in the stroma of uterine mesometrial pole,as round, immature and small lymphocyte-like cells.With the progesterone(P_4)administered, the immunostaining results showed that DAB-lectin staining were mainly distributed in the vascular endothelial cells.With the combination of E_2 and P_4, DAB-lectin staining was distributed in the matrix of the uterus,seen as a number of small round uNK cells or some as vascular endothelial cells.The effects could be completely abolished by specific antagonists of their nuclear receptors(estrogen and progesterone receptor).Conclusion: The distribution of uNK cells in mouse uteri is collaboratively regulated by estrogen and progesterone.The endometrial uNK cells may be involved in the mechanism of the fetal protective immune reaction during pregnancy.

Severe acute respiratory syndrome is an infectious disease caused by a new type of coronavirus. It belongs to the seasonal febrile diseases in traditional Chinese medicine. The prevention and treatment of severe acute respiratory syndrome (SARS) can be under the guidance of the doctrines for treating febrile diseases of traditional Chinese medicine, treatment based on syndrome differentiation, such as syndrome differentiation of triple energizer, syndrome differentiation according to defensive phase, qi phase, nutrient phase and blood phase. During April and May of 2003, 8 cases of SARS were diagnosed in Shanghai, and 6 patients accepted complementary therapy of traditional Chinese medicine, without death case. The only one patient who didn't take glucocorticoid therapy was complementarily treated with traditional Chinese herbs through the whole treating procedure. Upon the successful treatment of the eight cases of SARS in Shanghai, it is demonstrated that the triple-energizer syndrome differentiation and defensive-qi-nutrient-blood syndrome differentiation in traditional Chinese medicine are of high value in treating SARS patients.

Objective To screen the proteins with decreased expression in the synovial tissues of rheumatoid arthritis (RA) patients by comparing their expression profiles with that of osteoarthritis (OA) and ankylosing spondylitis (AS) patients by a proteomic approach,and to explore the association of reduced expression with disease susceptibility by a single nucleotide polymorphism (SNP) analysis.Methods Proteins extracted from the synovial membranes (n=10 for each disease) were separated by 2-D electrophoresis.The proteins with significantly decreased expression in the RA samples were subjected to MALDI-TOF-MS.The results were verified using Western blotting.Tag SNPs located in the targeted gene were assessed using the Taqman assay in a cohort of 267 Chinese patients with RA and 160 healthy controls.The genotyping results were confirmed in a large cohort of 389 patients with RA and 371 healthy controls.SPSS 11.5 software package was used for one way ANOVA and Fisher's exact test.Results The expression of vitamin D-binding protein (VDBP) in the synovial membranes from patients with RA was significantly decreased when examined by proteomic approach.This result was confirmed by Western blotting analysis.The rs2282679 was significantly associated with RA (P=0.026 794).This result was confirmed in a large cohort of RA( OR=0.678 639,95％CI 0.54l 113-0.851 118,P=0.000 776).Conclusion Compared with samples from patients with OA and AS,RA patients' synovial tissues have low VDBP expression when examined by the proteomic method.The tag SNP rs2282679 located in VDBP is significantly associated with RA.The decreased expression and the genetic effect of VDBP in RA suggest that a novel pathogenic pathway,in which vitamin D contributes,may be involved in the arthritis process of RA.

OBJECTIVE:To study the rational use of antibacterials in inpatients.METHODS:A discussion was made on the use of antibacterials for 4 948 inpatients in 2004 and 5 476 inpatients in 2005 in our hospital,managed in accordance with the requirements for hospital management by level and the Guidelines for Clinical Use of Antibacterials.RESULTS & CONCL- USIONS:The inpatient use of antibacterials in our hospital has been made basically rational through implementation of systematic management,enactment of antibacterials management by level,monitoring on usage of antibacterials and enforcement of warning system for overusing.

Objective To explore the optimal experiment conditions of CCK-8 and MTS for cell proliferation assays in human amniotic epithelial cells and to evaluate the cytotoxicity of these reagents. Methods Human amniotic epithelial cells (hAECs) in logarithm growth stages were prepared in different cell concentrations with DMEM/F12 and 10% FBS. The sensitivity and optimal wavelengths was determined based on the optical density (OD) measured at 450 nm and 492 nm. The optimal time was determined under the conditions of the same cell concentration and defined OD values. HAECs were treated with DMSO, CCK-8 and MTS for 1 h, 2 h, 3 h, and 4 h, respectively. 24 h later, cytotoxicity of the CCK-8 and MTS was evaluated by determination of cell proliferation and Trypan Blue staining. Results The optimal detection wavelength was 450 nm for CCK-8, and 492 nm for MTS. The sensitivity of CCK-8 was slightly lower then that of MTS. The optimal time for incubation hAECs with CCK-8 was 4 h within 1~4 h. The inhibitory on cell proliferation and cytotoxicity of CCK-8 were weaker then those of MTS. Conclusion CCK-8 is a convenient reagent with low cytotoxicity for detection of the proliferation of hAECs.

Objective:To explored the effect of 78c in treating collagen-induced arthritis (CIA) mice and to investigate its mechanism of effects.Methods:CIA mice model and CD38 +NK cells were treated with 78c. Cytokine concentrations and lymphocyte subtypes were measured in the mice peripheral blood and culture medium using flow cytometry. Mikenyi cell isolation kit was used to isolate CD4 + T cells and NK cells from peripheral blood mononuclear cells of healthy volunteers. CD38 + NK cells were enriched using the Miltenyi CD38 microbeads from the extracted NK cells. CD38 + NK cells with 78c pretreatment or not were cocultured with CD4 +T cells in transwells. The least significant difference (LSD) method was used for comparison between the two groups, and one-way analysis of variance was used for multi-group significance. Pearson correlation analysis was used for correlation analysis. Results:78c treatment significantly suppressed joint inflammation, inhibited the toe thickness of CIA mice, and reduced the number of while cell, neutrophils, platelets, and concentrations of IFN-γ, IL-6 and TNF-α ( t=6.10, P<0.001; t=4.00, P=0.002; t=3.09, P=0.012; t=2.31, P=0.043; t=3.58, P=0.005; t=2.68, P=0.002) in the CIA mice. The proportion of CD38 +NK cells decreased from (3.9±0.9)% to (2.4±0.3)% ( t=2.49, P=0.032), the proportion of regulatory T cell (Treg) increased from (0.81±0.33)% to (1.41±0.26)% ( t=2.74, P=0.021), and the concentration of IL-10 also increased from (99±37) pg/ml to (199±9) pg/ml( t=2.76 , P=0.020). The proportion of Treg in CD4 +T cells cocultured with 78c-pretreated CD38 +NK cells increased from (0.52±0.04)% to (0.69±0.08)% ( t=3.33, P=0.029) , the T helper cells (Th)17/Treg ratio decreased from (4.44±0.26) to (2.59±0.64) ( t=4.76 , P=0.009), and the Th1/Th2 ratio decreased from (14.8±1.6) to (8.1±1.3)( t=5.70 , P=0.005). Conclusion:78c can reduce the proportion of CD38 +NK cells, thereby reducing the inhibition of CD38 +NK cells on CD4 +T cell differentiation into Treg cells, leading to the restoration of immune balance. The results of this study suggest that 78c is a potential therapeutic agent for rheumatoid arthritis.

Objective:To investigate cases of delayed lactation initiation in women with transvaginal delivery and the influencing factors, in order to provide a basis for effective control of delayed lactation initiation and promotion of breastfeeding.Methods:Inpatients who were admitted to the obstetric ward of Affiliated Hospital of Jining Medical College from November 6, 2020 to January 16, 2021 were selected for the study using convenience sampling method and investigated by general information questionnaire and Chindbirth Experience Questionnaire (CEQ). Binary Logistic regression analysis was used to determine the factors influencing delayed lactation initiation.Results:The incidence of delayed lactation initiation in 622 women with transvaginal delivery was 38.75% (241/622). Binary Logistic regression analysis showed that age 20-35 years, full-term delivery, labor and delivery, use of labor analgesia, and good experience of transvaginal delivery were protective factors for delayed lactation initiation ( OR values were 0.012 to 0.868, all P<0.05); age >35 years, excessive weight gain during pregnancy, presence of pregnancy complications, use of induction of labor during delivery, long labor process, and damage to perineal skin after delivery were risk factors for delayed lactation initiation ( OR values were 1.097 to 13.235, all P<0.05). Conclusions:The high incidence of delayed lactation initiation in women with transvaginal delivery is influenced by a number of factors, which reminds the clinic that lactation in women after transvaginal delivery also needs to be taken into account, with priority assessment and prevention for those who are elderly (age≥35 years), primiparous, have other diseases during pregnancy, have gained too much weight during pregnancy, have preterm delivery, have a long duration of labor, have not received labor analgesia, have had a single or combined induction of labor, have had an episiotomy or perineal laceration during labor, and have a poor transvaginal delivery experience.

Objective:To evaluate the feasibility of 8E5 cells and CD19-CAR-Jurkat cells used as reference cells in the detection of lentiviral vector integration sites with different methods.Methods:Single clones of 8E5 cells and CD19-CAR-Jurkat cells were selected using limiting dilution method. Digital PCR was established to detect the copy number of HIV-1 in 8E5 cells and the copy number of CAR in CD19-CAR-Jurkat cells. High-throughput sequencing techniques (whole-genome resequencing, modified genome sequencing and probe hybridization capture) were used to detect integration sites in 8E5 cells and CD19-CAR-Jurkat cells, and optical genome mapping (OGM) technology was used for further confirmation.Results:Three clones of 8E5-D8 cells and six clones of CD19-CAR-Jurkat 2-6 cells were selected using the limiting dilution method. 8E5-D8 and CD19-CAR-Jurkat 2-6 were chosen as candidate cells based on their gene copy numbers detected by digital PCR and flow cytometry. These cells were then expanded and cryopreserved. Digital PCR showed that 8E5-D8 cells contained approximately 1 copy per cell, while CD19-CAR-Jurkat 2-6 cells contained approximately 13 copies per cell. High-throughput sequencing revealed one integration site in 8E5 cells and 13 integration sites in CD19-CAR-Jurkat cells, which matched the copy number detection results. All these integration sites were further confirmed at the submicroscopic level of chromosomes using OGM.Conclusions:Based on the insertion copy numbers and integration sites, 8E5-D8 cells and CD19-CAR-Jurkat 2-6 cells could be used as reference cells in further development of methods for detecting integration sites in CAR-T cell lentiviral vectors.