OBJECTIVE: To investigate the effectiveness and preliminary mechanisms of icariin (ICA) in enhancing the reparative effects of adipose-derived stem cells (ADSCs) on skin radiation damagies in rats. METHODS: Twelve SPF-grade Sprague Dawley rats [body weight (220±10) g] were subjected to a single dose of 10 Gy X-ray irradiation on a 1.5 cm×1.5 cm area of their dorsal skin, with a dose rate of 200 cGy/min to make skin radiation damage model. After successful modelling, the rats were randomly divided into 4 groups ( n=3), and on day 2, the corresponding cells were injected subcutaneously into the irradiated wounds: group A received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL), group B received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL)+1 μmol/L ICA (0.1 mL), group C received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL) pretreated with a hypoxia-inducible factor 2α (HIF-2α) inhibitor+1 μmol/L ICA (0.1 mL), and group D received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL) pretreated with a Notch1 inhibitor+1 μmol/L ICA (0.1 mL). All treatments were administered as single doses. The skin injury in the irradiated areas of the rats was observed continuously from day 1 to day 7 after modelling. On day 28, the rats were sacrificed, and skin tissues from the irradiated areas were harvested for histological examination (HE staining and Masson staining) to assess the repair status and for quantitative collagen content detection. Immunohistochemical staining was performed to detect CD31 expression, while Western blot and real-time fluorescence quantitative PCR (qRT-PCR) were used to measure the protein and mRNA relative expression levels of vascular endothelial growth factor (VEGF), platelet-derived growth factor BB (PDGF-BB), fibroblast growth factor 2 (FGF-2), interleukin 10 (IL-10), transforming growth factor β (TGF-β), HIF-2α, and Notch1, 2, and 3. RESULTS: All groups exhibited skin ulcers and redness after irradiation. On day 3, exudation of tissue fluid was observed in all groups. On day 7, group B showed significantly smaller skin injury areas compared to the other 3 groups. On day 28, histological examination revealed that the epidermis was thickened and the dermal fibers were slightly disordered with occasional inflammatory cell aggregation in group A. In group B, the epidermis appeared more normal, the dermal fibers were more orderly, and there was an increase in new blood vessels without significant inflammatory cell aggregation. In contrast, groups C and D showed significantly increased epidermal thickness, disordered and disrupted dermal fibers. Group B had higher collagen fiber content than the other 3 groups, and group D had lower content than group A, with significant differences ( P<0.05). Immunohistochemical staining showed that group B had significantly higher CD31 expression than the other 3 groups, while groups C and D had lower expression than group A, with significant differences ( P<0.05). Western blot and qRT-PCR results indicated that group B had significantly higher relative expression levels of VEGF, PDGF-BB, FGF-2, IL-10, TGF-β, HIF-2α, and Notch1, 2, and 3 proteins and mRNAs compared to the other 3 groups ( P<0.05). CONCLUSION ICA may enhance the reparative effects of ADSCs on rat skin radiation damage by promoting angiogenesis and reducing inflammatory responses through the HIF-2α-VEGF-Notch signaling pathway.
Animals ; Rats, Sprague-Dawley ; Skin/pathology* ; Rats ; Vascular Endothelial Growth Factor A/genetics* ; Basic Helix-Loop-Helix Transcription Factors/genetics* ; Signal Transduction ; Flavonoids/pharmacology* ; Adipose Tissue/cytology* ; Stem Cells/cytology* ; Receptors, Notch/metabolism* ; Radiation Injuries, Experimental/metabolism* ; Wound Healing/drug effects* ; Male

OBJECTIVE To investigate the effect of pachymic acid （PA） on renal function and fibrosis in chronic renal failure （CRF） rats and its potential mechanism based on the Ras homolog gene family member A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） signaling pathway. METHODS Using male SD rats as subjects， the CRF model was established by 5/6 nephrectomy； the successfully modeled rats were divided into model group， PA low-dose， medium-dose and high-dose groups （5， 10， 20 mg/kg PA）， high-dose PA+ROCK pathway activator lysophosphatidic acid （LPA） group （20 mg/kg PA+1 mg/kg LPA）， with 15 rats in each group. Another 15 rats were selected as the sham operation group with only the kidney exposed but not excised. The rats in each drug group were gavaged and/or injected with the corresponding liquid via the caudal vein， once a day， for 12 consecutive weeks. During the experiment， the general condition of rats was observed in each group. After the last administration， the serum renal function indexes （blood urea nitrogen， serum creatinine， uric acid） of rats in each group were detected， the renal histopathological changes were observed； the renal tubule injury score and the area of renal fibrosis were quantified. The levels of oxidative stress indexes [malondialdehyde （MDA）， superoxide dismutase （SOD）] and inflammatory factors （tumor necrosis factor-α， interleukin-1β， interleukin-6）， the positive expression rates of connective tissue growth factor （CTGF） and collagen Ⅰ were detected as well as the expression levels of pathway-related proteins （RhoA， ROCK1） and fibrosis- related proteins （transforming growth factor-β1， bare corneum homologs 2， α-smooth muscle actin） were determined. RESULTS Compared with the sham operation group， the rats in model group had reduced diet， smaller body size， listless spirit and sluggish response， reduced and atrophied glomeruli， dilated renal tubules with chaotic structure， and a large number of inflammatory cells infiltrated interstitium； the serum levels of renal function indexes， renal tubule injury score， renal fibrosis area proportion， the levels of MDA and inflammatory factors， the positive expression rates of CTGF and collagen Ⅰ， and the expression levels of pathway-related proteins and fibrosis-related proteins in renal tissues were significantly increased， while SOD level was significantly decreased （P＜0.05）. Compared with the model group， the general condition and pathological injuries of kidney tissue of rats in PA groups were improved to varying degrees，and the above quantitative indexes were significantly improved in a dose-dependent manner （P＜0.05）. LPA could significantly reverse the improvement effect of PA on the above indicators （P＜0.05）. CONCLUSIONS PA can improve renal function and alleviate renal fibrosis in CRF rats， which may be related to inhibiting the activation of RhoA/ROCK signaling pathway.

OBJECTIVE To investigate the effect and mechanism of Astragalus polysaccharide （APS） on peritoneal fibrosis and angiogenesis in rats with peritoneal dialysis （PD）. METHODS Rats were randomly divided into normal control group （Control group）， model group （PD group）， 70 mg/kg APS group （APS-L group）， 140 mg/kg APS group （APS-H group）， and 140 mg/kg APS+40 mg/kg hypoxia-inducible factor-1α （HIF-1α） agonist DMOG group （APS-H+DMOG group）， with 12 rats in each group. PD rat models were constructed in the last four groups of rats. Administration groups were given APS intragastrically and DMOG intraperitoneally. Control group and PD group were given constant volume of normal saline intragastrically， once a day， for 4 consecutive weeks. After the last medication， the peritoneal ultrafiltration （UF）， mass transfer of glucose （MTG）， the levels of serum creatinine （Scr） and blood urea nitrogen （BUN） were detected in rats； peritoneal histomorphology and peritoneal fibrosis （peritoneal thickness and proportion of collagen fiber deposition） were observed； the microvascular density and the expression levels of α-smooth muscle actin （α-SMA）， laminin （LN）， HIF-1α and vascular endothelial growth factor （VEGF） proteins were detected in peritoneal tissue of rats. RESULTS Compared with Control group， the mesothelium of rats in the PD group was loosely arranged and shed， inflammatory cells infiltrated， the peritoneal thickness and proportion of collagen fiber deposition were increased significantly （P＜0.05）. The levels of MTG， Scr and BUN in serum， microvascular density and the expressions of α-SMA， LN， HIF-1α and VEGF proteins were significantly increased， while the level of UF was significantly decreased （P＜ 0.05）； compared with PD group， the levels of above indexes were significantly reversed in APS-L and APS-H groups （P＜0.05）， and the improvement of APS-H group was better than APS-L group （P＜0.05）. Compared with APS-H group， the levels of above indexes in APS-H+DMOG group were all reversed （P＜0.05）. CONCLUSIONS APS inhibits peritoneal fibrosis and angioge-nesis in PD rats by inhibiting HIF-1α/VEGF signaling pathway.

Objective: To investigate the clinical manifestations, pathological features, and treatment of oral and maxillofacial pyogenic granulomas induced by camrelizumab. Methods: A case of pyogenic granuloma of the gums and lips caused by camrelizumab was reported along with a literature review. Results: After 4 months of treatment with camrelizumab for liver cancer, the patient developed systemic reactive capillary hyperplasia (RCH), followed by multiple masses on the lower lip and gingiva. After periodontal therapy, the masses on the lower lip and the gingiva were removed, and camrelizumab administration was stopped. The pathological result was gingival pyogenic granuloma/granulomatous hemangioma. No new masses were found in the oral cavity during postoperative follow-up. A review of the literature showed that RCH is the most common adverse drug reaction to camrelizumab but it occurs infrequently in the oral cavity. At present, the etiology of RCH has not been clarified, but the research has shown that camrelizumab may trigger tissue proliferation into hemangiomas by activating vascular endothelial cells, and the combined use of camrelizumab is safer than single use. RCH is self-limiting and most cases resolve spontaneously after discontinuation of the drug. If the mass causes dysfunction, surgical excision is feasible. Conclusion Camrelizumab can cause oral and maxillofacial reactive capillary hyperplasia complicated by pyogenic granuloma.

Objective:To explore the application value of sevoflurane inhalation anesthesia in mitral valve replacement.Methods:A total of 94 patients who underwent mitral valve replacement in our hospital (October 2016-October 2018) were randomly divided into the control group ( n=47) and the observation group ( n=47). The control group received target-controlled infusion of propofol, and the observation group inhaled sevoflurane.The postoperative conditions [intensive care unit (ICU) stay time, extubation time of tracheal tube, spontaneous cardiac rebound], hemodynamic indexes [mean arterial pressure (MAP), heart rate (HR)], serum creatine phosphokinase isoenzyme (CK-MB), cardiac troponin I (cTnI), malondialdehyde (MDA) and superoxide dismutase (SOD) in the two groups were analyzed. The patients were followed up for one month. The incidence of major adverse cardiovascular events (MACE) was calculated. Results:⑴ Postoperative situation: the time of stay in ICU and extubation of tracheal tube in the observation group was shorter than that in the control group, and the rate of spontaneous cardiac rebound (93.62%) was higher than that in the control group (72.34%) ( P<0.05); ⑵ Hemodynamic index level: there was no statistically significant difference in MAP and HR levels between two groups before operation, before cardiopulmonary bypass, after cardiopulmonary bypass, and after operation ( P>0.05); ⑶ CK-MB and cTnI: the levels of serum CK-MB and cTnI in the two groups were higher at 2, 6, 24, and 48 h after aortic cross-clamp release than before anesthesia induction, but the indicators of the observation group were lower than those in the control group; ⑷ MDA and SOD: the serum SOD level in the two groups at 2, 6, 24, and 48 h after aortic cross-clamp release were lower than before anesthesia induction, and the MDA level in the two groups at 2, 6, 24, and 48 h after aortic cross-clamp release were higher than before anesthesia induction. The level of SOD in the observation group was higher than that in the control group, and the level of MDA was lower than that in the control group ( P<0.05); ⑸ MACE: the incidence of MACE in the observation group (12.77%) was lower than that of the control group (29.79%) ( P<0.05). Conclusions:During mitral valve replacement, sevoflurane inhalation anesthesia can maintain hemodynamic stability. The duration of ICU stay and tracheal tube extubation time is shorter, and the fluctuation of serum CK-MB, cTnI, MDA and SOD is small, and it can reduce the risk of MACE.

Aim To investigate the effect of genistein on apoptosis in triple-negative breast cancer MDA-MB-231 cells and the underlying mechanisms.Methods MTT assay was used to detect the inhibition rate on breast cancer MDA-MB-231 cells of genistein.Hoechst 33258 staining was applied to determine the effect of genistein on morphology of MDA-MB-231 cells.qRT-PCR was employed to detect the mRNA expression of EGFR in MDA-MB-231 cells.Western blot was utilized to determine the expression of Bcl-2, Bax, caspase-3, EGFR, Akt, and p-Akt.The expressions of Akt and p-Akt proteins in breast cancer MDA-MB-231 cells were detected after treated with Akt activator insulin, genistein and in combination with insulin.Results Genistein inhibited the viability of breast cancer MDA-MB-231 cells in a time-dependent manner.The results of Hoechst 33258 staining showed a typical apoptotic morphological changes of MDA-MB-231 cells after treatment of genistein for 36 h.qRT-PCR showed that the mRNA expression of EGFR in MDA-MB-231 cells decreased after treated with genistein for 36 h.The expression levels of Bcl-2, EGFR, Akt, p-Akt, ERK, p-ERK were significantly down-regulated(P<0.01) compared with control.While, the expression of Bax, caspase-3 was significantly up-regulated (P<0.01).It was observed that p-Akt was significantly activated after the treatment of Akt activator insulin (P<0.01), however, significantly down-regulated (P<0.01) when treated with genistein.Conclusion Genistein could inhibit the growth of triple-negative breast cancer MDA-MB-231 cells and induce apoptosis, which probably involves regulating EGFR/PI3K/Akt signaling pathway.

Objective To evaluate the role of calcium channel in the mechanism of the generation and maintenance of bursting firing of substantia nigra pars compacta (SNc) dopaminergic neurons in rats.Methods Using the patch clamp technique,we observed the firing pattern switching features after adding 10 μmol/L N-methyl-D-aspartic acid (NMDA),compared the changes of whole-calcium current and L-type calcium current with or without NMDA,and analyzed the correlation between the generation of burst firing and L-type calcium channel activation.Results After NMDA treatment,the firing pattern of SNc dopaminergic neurons changed to burst firing,which was compromised by a charastistic high plateau potential and series of action potential on it.The current density of L-type calcium current increased significantly after adding NMDA,which,from (2.86 ±0.26) pA/pF (n =28),significantly increased to (3.75 ± 0.18) pA/pF (n =34 ; t =7.52,P =0.002 8).The high plateau potential was almost abolished with the application of verapamil,a specific antagonist of L-type calcium channel.Consiusion NMDA could induce the firing pattern changed to burst firing in SNc dopaminergic neurons,while L-type calcium channel contributes to the process of generation and maintenance of burst firing.

Objective To explore the effects of early food intake on recovery of patients with postoperative upper urinary tract. Methods One hundred and twenty-six patients after upper urinary tract surgery from January to December 2012 were divided into early food intake group ( food intake beginning 6 h after surgery) and conventional group ( food intake began after first flatus ) . The recovery time of gastrointestinal function, hospital stay, drainage volume and other parameters in two groups were compared. Results The time of flatus, leaving the sickbed, defecation, stopping transfusion and getting right time of eating were (1. 80 ± 0. 46), (1. 45 ± 0. 51), (4. 06 ± 1. 10), (3. 27 ± 0. 85), (3. 33 ± 1. 16) d in the early food intake group better than those of the conventional group (t=3. 598, 2. 239, 4. 722, 3. 140, 3. 395;P<0. 05). Conclusions Early food intake after surgery of upper urinary tract can promote recovery of gastrointestinal function and decrease fluid infusion and hospital stay.

Objective To improve diagnose and therapy of post-traumatic acute renal failure induced by rhabdomyolysis. Methods A total of 20 patients with post-traumatic acute renal failure induced by rhabdomyolysis were analyzed retrospectively in aspects of clinical manifestation, laboratory examination and treatment. Of all, there were 9 patients treated with continuous renal replacement therapy (CRRT), while the other 11 were set as control, receiving no CRRT. Results After treatment with CRRT, 7 patients obtained clinical curing, with 2 deaths. As for control patients, there were 5 patients with secondary chronic renal insufficiency, 1 with clinical curing and 5 deaths. Conclusions Early diagnosis, CRRT, sufficient hydration, hematedialysis and supportive treatment are key points to improve the cure rate. It is important to apply CRRT for patients with renal inadequacy.

OBJECTIVE To discover diversity of X gene sequences of hepatitis B virus isolates in hepatitis B induced liver cirrhosis patients and HBV carriers.METHODS DNA fragment including X gene sequences of hepatitis B virus was amplified,sequencing PCR products was preformed.The PCR products of three liver cirrhosis patients and three HBV symptomless carriers were cloned into pGEMT Easy vectors.Positive clones with target sequences were selected out for sequencing.Sequence comparison was made to find the identity.RESULTS A comparison of T1762/A1764,G1719T,T1727G/A,G1730C and T1753C mutations in a core promoter between the liver cirrhosis patients and the HBV carriers showed that the HBV isolates from the former had higher frequencies of mutation than the latter.The X promoter region of the HBV isolates from the liver cirrhosis patients showed higher frequencies of mutation than the isolates from the HBV carriers.Additionally,the homology between clones of X gene from one individual with liver cirrhosis averages 91.3-99.7%,that of HBV carriers averages 96-100%.CONCLUSIONS The core and X promoter region of the HBV isolates from the liver cirrhosis patients show the higher frequencies of mutation than the isolates from the HBV carriers.There are HBV quasispecies which possess great variation in the liver cirrhosis patients.