Objective: To determine the effect of a recombinant lentivirus containing human stem cell
leukemia (SCL) gene on the expression of c-kit protein in damaged interstitial cells of Cajal (ICC)
under high glucose condition.
Methods: Aft er isolation of ICC, the cells were cultured for 24 hours until the cells were adherent.
Aft er identifi cation by inverted microscope and immunofl uorescence, ICC cells were divided into two groups: A control group and a high glucose group. The control group was added with a medium
containing 5 mmol/L of glucose. The high glucose group was added with a medium containing
20 mmol/L of glucose. After 48 h of continuous cultivation, the high glucose group was divided
into 3 subgroups: A blank group, an empty lentivirus group, and an experimental group. The blank
group, the empty lentivirus group, and the experimental group were added a medium containing
PBS solution, empty lentivirus, and a recombinant lentivirus containing the SCL gene with a
glucose concentration of 5 mmol/L, respectively. The cultures were incubated for 24 and 48 h. The
expression of c-kit protein in ICC in each group was detected by Western blot.
Results: After 24 or 48 h, the expression of c-kit protein in ICC was significantly lower in the blank
group and the lentivirus group than that in the control group, and the expression of c-kit protein in
ICC was significantly higher in the experimental group than that in the blank group and the empty
lentivirus group, but it was still lower than that in the control group (all P<0.05).
Conclusion: The recombinant lentivirus of SCL gene can up-regulate the expression of c-kit
protein in functionally impaired ICC under high glucose condition.
Objective: To clone human mitogen-activated protein kinase kinase 6 (MKK6) gene promoter and
explore its transcription activity by ubiquitin specifi c peptidase 22 (USP22).
Methods: MKK6 gene promoter was amplified by PCR and two bases mutation within USP22
binding site was subsequently introduced. The wild type and mutant MKK6 promoter were
inserted into the luciferase report vector pGL3-Basic, respectively. Recombinant plasmids were cotransfected
with plasmid pRL-TK into HeLa cells, and the luciferase activities were measured by
dual luciferase reporter system. Furthermore, the direct interaction between USP22 and MKK6 promoter was detected by chromatin immunoprecipitation (ChIP) assay. Finally, the MKK6
transcription activity was measured after knockdown of USP22.
Results: The recombinant luciferase report vectors containing wild or mutant type of MKK6
promoter were successfully constructed. Mutation of USP22 binding site resulted in decrease of
MKK6 promoter-driven luciferase activity in HeLa cells (P<0.05). USP22 could interact directly
with MKK6 promoter. Down-regulation of USP22 led to the decreased MKK6 mRNA expression
Conclusion: USP22 could regulate the transcription activity of MKK6 gene in HeLa cells.
Objective: To investigate the effect of prophylactic aucubin (AU) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice.
Methods: Male BABL/c mice were randomly divided into a control group, an ALI group, and
an AU treatment group, 16 mice in each group. ALI mice were injected with LPS (5 mg/kg,
intratracheal injection), and AU (10 mg/kg) was injected intraperitoneally 30 min ahead. After LPS
injection for 6 hours mice were sacrificed, the morphological changes of lung tissues were detected
by HE staining and the lung injury score was obtained. The mRNA expression of tumor necrosis
factor-α (TNF-α) and interleukin 10 (IL-10) in lung tissue was detected by real-time PCR. The
total protein and lactate dehydrogenase (LDH) activity, the cell count, and the protein content of
TNF-α and IL-10 in the mouse bronchoalveolar lavage fluid (BALF) were detected.
Results: Compared with ALI mice, the pathological damage score of lung tissue was significantly
reduced in the AU group, the total number of BALF cells, neutrophils, and macrophages were
significantly decreased, LDH activity and the total protein content were also significantly decreased
(all P<0.01). In addition, AU can reduce the mRNA and protein expression of TNF-α in lung of
ALI mice, and increase the mRNA and protein expression of IL-10 (all P<0.01).
Conclusion: AU can reduce LPS-induced ALI in mice.
Objective: To investigate the effects of airway epithelial cells on macrophages chemotaxis and
infl ammatory cytokine expression under hypoxic conditions.
Methods: Human bronchial epithelial cells (HBE) treated with diff erent concentrations (0, 100, 200, 400, 800 μmol/L) of CoCl2 or transfected with HIF-1α siRNA were co-cultured with THP-
1-derived M1 macrophages or M2 macrophages. The chemotactic effects on macrophages were
analyzed by Transwell assay. The levels of TNF-α, IFN-γ, IL-4, IL-13 and IL-10 in the supernatants
of macrophages were detected by ELISA, and HIF-1α or Cav-1 mRNA expression in HBE or
macrophages was detected by RT-qPCR.
Results: HBE cells promoted macrophages chemotaxis in a time- and concentration-dependent
manner. Compared to un-transfected group, the chemotactic ability of HBE transfected with HIF-
1α siRNA was significantly weakened (P<0.01). Under the same culture conditions, the chemotaxis
of M2 macrophages was greater than that in THP1-derived M1 macrophages. The concentrations of
TNF-α, IFN-γ, IL-4, IL-13 and IL-10 in the supernatants of macrophages were increased in a timeand
concentration-dependent manner. The concentrations of TNF-α and IFN-γ were increased
further after co-culturing for 8 and 12 h; while IL-4, IL-13 and IL-10 concentrations were increased
further during 24 h of co-culture. The levels of cytokines in the supernatants of macrophages cocultured
with HBE and transfected with HIF-1α siRNA were significantly lower than those in untransfected
cells (P<0.05 or P<0.01). The reduction of TNF-α or IFN-γ was more obvious. The
expression of HIF-1α or Cav-1 mRNA in HBE or macrophages was increased in a concentrationdependent
manner after 8 or 12 h co-culture, which was significantly reduced when HBE was
transfected with HIF-1α siRNA.
Conclusion: Airway epithelial cells can enhance macrophages chemotaxis and pro-inflammatory
cytokines expressions under hypoxic condition. HIF-1α and Cav-1 may be the important mediators
in these processes.
Objective: To investigate the eff ects of chemerin on helper T cells 9 (Th 9)/regulatory T cells (Treg)
in patients with psoriasis and the potential molecular mechanisms.
Methods: Twenty-fi ve patients with psoriasis and twenty healthy volunteers were selected for this
study. CD4+ T cells were isolated from peripheral blood of samples by magnetic bead separation.
Th e levels of chemerin and its receptor chemR23 were detected by real-time RT-PCR and ELISA.
CD4+ T cells isolated from the healthy volunteers were treated with different concentrations of
chemerin (50, 100, 150, 200 ng/mL), then cell viability was detected by MTT assay. Th e expression
of infl ammatory molecules and Th 9/Treg were detected by ELISA and fl ow cytometry, respectively.
Results: The expressions of chemerin and chemR23 in peripheral blood from patients with psoriasis were higher than those in healthy control (both P<0.05). The Th9/Treg was higher in
patients with psoriasis than that in healthy control (P<0.05). After treating CD4+ T cells with
150 ng/mL of chemerin, the levels of IL-6, IL-9 and IL-17 were increased significantly (all P<0.05).
Additionally, Th9/Treg was increased (P<0.05) and the cell balance was disrupt. However, the
effects of chemerin on CD4+ T cells were reversed by silencing of chemR23 (all P<0.05).
Conclusion: Chemerin may regulate the immune balance for Th9/Treg in CD4+ T cells from
patients of psoriasis.
Objective: To investigate the eff ect of adipose-derived stem cells (ADSCs) on radiation-induced skin injury in SD rats.
Methods: Radioactive particles 192Ir were used to irradiate the left medial thigh skin of SD rats,
and the irradiation dose was at 90 Gy. Then, the rats were randomly allocated into a control group
and a treatment group (each n=9). After the irradiation, the control group was injected with
60 μL PBS and the treatment group was injected with 60 μL ADSCs in irradiated skin. The progress
of skin damage and healing was observed and photographed every day. Twenty-eighth days
after the irradiation, the irradiated skin tissue was taken from the left thigh, and then fixed with
formaldehyde fixative solution. At the same time, the skin tissue of the corresponding part of the
normal group (n=9) that was not irradiated was also taken. After sampling, embedding and slicing,
immunohistochemical staining was used to compare the levels of α-smooth muscle actin (α-SMA),
and HE staining was used to compare pathological features of the skin.
Results: Radioactive particle 192Ir caused the development of III or IV radioactive skin damage.
The score of the treatment group was significantly lower than that of the control group. The wounds
of the treatment group were basically healed at 28 days, while the ulcer of the control group was
unhealed. So, the healing time was shorter in the treatment group. The expression of α-SMA in
the skin of the two groups was increased after the radiotherapy. By analyzing the pathological
microstructure image, we found that the thickness of epidermis in the control group was greater
than that in the treatment group, while the vascular density in the treatment group was greater than
that in the control group (all P<0.05).
Conclusion: Radioactive particles 192Ir can cause skin damage, while the adipose-derived stem cells
might alleviate radiation-induced skin injury and promote ulcer healing by promoting angiogenesis.
Objective: To compare the cumulative live birth rates (CLBR) and the incidence of ovarian
hyperstimulation syndrome (OHSS) between fresh embryo transfer (ET) and frozen ET (the
freeze-all policy), when oocyte numbers are more than 15 in the first treatment of in vitro
fertilization or intracytoplasmic sperm injection, and to evaluate the benefi ts of the freeze-all policy.
Methods: We retrospectively analyzed clinical data of 2 842 patients whose oocytes numbers were
more than 15, including 1 095 frozen ET patients and 1 747 fresh ET patients. Th e patients general
data, a baseline features, CLBR, and the incidence of OHSS were compared between the 2 groups.
Results: There were 598 patients in the 2 groups after they experienced the propensity score
matching. No significant differences were found in age, infertility causes, body mass index, basal
follicle stimulating hormone level, the total days and total dose of using gonadotrophin (Gn)
between the 2 groups (all P>0.05). The CLBR of the freeze-all cycles increased along with the
number of oocytes (P>0.05), and the oocyte numbers were greater in freeze-all group than those
of the fresh ET group (P<0.001). There was no significant difference in CLBR after one complete
cycle between the 2 groups (P>0.05), but after the first embryo transfer cycle, the CLBR in freezeall
group was higher than that in the fresh ET cycle group (P<0.05). The incidence of OHSS in
patients with freeze-all was significantly lower than that in the patiants with fresh ET (P<0.05).
Conclusion: Patients with oocytes over 15 and OHSS tendency who accepted the freeze-all
strategy can help them to prevent OHSS and they have a higher CLBR than fresh ET cycles.