BACKGROUND: Clinically, in patients undergoing hematopoietic stem cell transplantation (HSCT), human cytomegalovirus (HCMV) can be associated with delayed platelet engraftment, phenotypically abnormal peripheral blood leukocytes, and graft rejection, possibly through a direct viral effect on hematopoietic progenitor cells after HCMV infection. OBJECTIVE: To investigate the inhibitory effect of ganciclovir (GCV) on proliferation of colony forming unit (CFU) granulocyte-macrophage (CFU-GM), CFU-erythroid (CFU-E), CFU T-lymphocyte (CFU-TL), CFU-multipotential (CFU-Mix) and CFU-megakaryocyte (CFU-Mk) progenitor cells of cord blood (CB) and the protective effects on them. DESIGN: Contrast observational study.SETTING: Department of Molecular Biology, Affiliated Hospital of Luzhou Medical College.PARTICIPANTS: A total of 20 cord blood (CB) samples (with 10 mL for each sample) from fetal umbilical vein of normal term spontaneous delivery neonates were provided by the Department of Gynaecology and Obstetrics, Affiliated Hospital of Luzhou Medical College. All the patients were informed and agreed with the experiment.METHODS: The experiment was carried out in the Department of Molecular Biology, Affiliated Hospital of Luzhou Medical College from June 2004 to December 2006. Colony forming unit-assay was applied to observe the suppression effect of HCMV-AD169 strain on CFU-GM, CFU-E, CFU-TL, CFU-Mix and CFU-Mk of CB with the presence of GCV. The techniques of polymerase chain reaction (PCR) and fluorescence quantification PCR were used to demonstrate the existence of HCMV-AD169 DNA in the colony cells of cultured CFU-GM, CFU-E, CFU-TL, CFU-Mix and CFU-Mk. Normal progenitor cells culture system was regarded as blank control group; normal progenitor cells culture system with inactivated HCMV fluid as inactivated (IV) control group.MAIN OUTCOME MEASURES: ① The number and maintaining duration of colonies of cultured progenitor cells were counted by using a light inverted phase contrast microscope. ② The techniques of PCR and fluorescence quantification PCR were used to demonstrate the existence of HCMV-AD169 DNA in the colony cells of cultured progenitor cells.RESULTS: ① Number and lasting time of colonies: The numbers of CFU-GM, CFU-E, CFU-TL, CFU-Mix and CFU-Mk colonies in the HCMV infection group were significantly less than those in the blank control group (P < 0.01). The maintaining duration of colonies in the HCMV infection group was significantly shorter than that in the blank control group (P < 0.01). HCMV-DNA copies of colony cells of GCV group decreased significantly by using fluorescence quantification PCR compared with HCMV group (P < 0.01), while negative in blank control and inactivated control in CFU-MK and CFU-Mix. ② CFU Growth rate: The Growth rate of colonies was 37.4%, 74.2%, 40.1%, 67.4% and 38.9% of CFU-GM, CFU-E, CFU-TL, CFU-Mix and FU-MK, respectively. ③ CFU-HCMV-AD169 DNA: Fluorescence quantification PCR showed that nucleonic acid content of progenitor cells after GCV-affected HCMV infection was decreased as compared with that after HCMV infection (P < 0.01).CONCLUSION: The differentiation and proliferation of CFU-GM, CFU-E, CFU-TL, CFU-Mix and CFU-Mk are significantly inhibited after infected with CMV-AD169 strain. The growth of hematopoietic progenitor cell after HCMV-AD169 infection is promoted by GCV, which suggests that GCV has an effect of anti-HCMV in vitro.

AIM: To establish a method for the determination of ephedrine hydrochloride in Juyuanzhike Tablets (Radix Platycodonis,Radix Polygalae,ephedrinelydrochloride,etc.). METHODS :Ephedrine hydrochloride in Juyuanzhike Tablets were determined by HPLC. RESULTS : The linear range was 0.2～1.6 ?g,r =0.9999.The average recovery was 97.7% and RSD was 0.58%,respectively. CONCLUSION : The method is simple,feasible and reproducible. It can be used for the quality control of Juyuanzhike Tablets.

Mice were divided into 3 groups:heavy infection group with 80 mice each was fed with 400 muscle larvae of Trichinella spiralis,light infection group with 60 mice each was fed by 200 larvae,and uninfected control (60 mice). The content of Cu,Zn and Fe in the dorsal hair samples was measured in the week of 1,3,5,7,9,11,13 and 15 after infection. Results indicated that the content of Zn,Cu and Fe in the two experimental groups reduced considerably in comparison to the control(P

OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) on ureteral injury repair via renal artery transplantation. METHODS: The left ureteral obstruction model was set up in 49 Balb/c mice by micro vascular clamp. The microscopic vascular clamp was taken out to lift the left ureteral obstruction after 10 days. The mice were randomly divided into an experimental group (n=25) and a control group (n=24). Balb/c mice BMSCs transfected by luciferase (Luc) were transplanted immediately through the renal artery after removing the microscopic vascular clamp from the experimental group; while mice in the control group was closed the incision after the microscopic vascular clamp was removed immediately and without BMSCs transplant. Magnetic resonance imaging (MRI) was used to scan the experimental mice. By measuring the left renal pelvis volume of the experimental mice at different time points and comparing the left ureter recanalization rate after removing left ureteral obstruction of the experimental group and the control group, we evaluated the repair effect of BMSCs on ureteral injury. RESULTS: The volume of the left renal pelvis in experimental mice became bigger obviously after the left ureter was obstructed (P<0.01). The left renal pelvis volume of the experimental group and the control group had no statistical significance 10 days after the left ureteral obstruction was set up (P=0.693). In the experimental group, the left ureter recanalization rate was higher than that in the control group, after removing the left ureteral obstruction (P=0.012). CONCLUSION Transplantation through the renal artery can promote the restoration of ureteral injury in mice.
Animals ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic Stem Cells ; cytology ; Mice ; Mice, Inbred BALB C ; Renal Artery ; Ureteral Obstruction ; therapy

Objective: To evaluate the quality status of piroxicam tablets. Methods: The samples were examined by the statutory standard,and the exploratory studies were carried out. The results were statistically analyzed. Results: Totally 138 batches were exam-ined according to the statutory standard, and among them, 135 batches were qualified with the qualified rate of 97. 8% . The unquali-fied item of 3 unqualified batches was dissolution. The exploratory studies showed that there were two crystal forms of piroxicam used in the tablets, and the dissolution of the two crystal forms was different with form 1 less than form 2. An inspection method for the relative substance was established. Totally 14 impurities were detected out and the structures of 8 impurities were identified. The impurities were mainly derived from the raw materials, and many batches of samples were with single largest impurity content exceeding 0. 5% , and the total of impurity content above 1. 0% . A class I solvent 1,2-dichloroethane was detected out in 13 batches of tablets by GC and confirmed by GC-MS. Through the dissolution consistency test, it was found that there was a great difference in the dissolution behavior among the products from different manufacturers. Conclusion: The overall quality of piroxicam tablets is not ideal, and the production process of some manufacturers needs to be improved.

BACKGROUND:Clinical skin wound healing continues to be a significant concern,and tissue repair research has moved to the forefront with the development of biomaterials with immunomodulatory properties.Therefore,it is crucial to research wound dressings that have immunomodulatory properties. OBJECTIVE:To prepare chitosan hydrogels that have been modified by arginine with different configurations and assess their capacity to speed up wound healing in a rat animal model. METHODS:(1)In vitro trial:Chitosan modified by pure L-arginine,pure D-arginine,and L-arginine and D-arginine was synthesized by EDC/NHS system,which was then crosslinked with aldehyde-modified four-arm polyethylene glycol.Different chitosan-based hydrogels(CS-L,CS-D,and CS-DL)were finally formed via the Schiff base reaction.Three kinds of hydrogel extracts were co-cultured with fibroblasts respectively.Hydrogel cytocompatibility was assessed using the CCK-8 assay and live/dead staining.The effect of hydrogel on the migration capacity of fibroblasts was assessed by using a scratch test.Three kinds of hydrogels were incubated with rat erythrocyte suspension respectively to evaluate the hemocompatibility of the hydrogels.The hydrogel extract was co-cultured with RAW264.7 macrophages to test the hydrogels'capacity to enhance macrophage NO generation and polarize macrophage phenotype.(2)In vivo experiment:A total of 36 adult SD rats were divided into 4 groups with 9 rats in each group by the random number table method.Two full-layer skin defect wounds of 2 cm×2 cm were made on the back of each rat.Normal saline was added to the wounds of the control group,and corresponding hydrogel was added to the wounds of the CS-L,CS-D,and CS-DL groups,respectively,and then bandaged and fixed.The wound healing was observed regularly after operation.Hematoxylin-eosin staining was performed at 3,10,and 21 days after operation.The samples were collected 10 days after operation and M2 macrophage immunofluorescence staining was performed. RESULTS AND CONCLUSION:(1)In vitro experiments:Under scanning electron microscopy,the three kinds of hydrogels exhibited obvious interpenetrating network structures with pore sizes ranging from 70-200 μm.The three kinds of hydrogels have good swelling performance,degradation performance,self-healing performance,and suitable mechanical strength.The three kinds of hydrogels had good cytocompatibility and hemocompatibility and could promote the migration of fibroblasts.All three kinds of hydrogels had the ability to promote the polarization of macrophages,and CS-D hydrogels had the strongest ability to promote the polarization of macrophages.CS-L hydrogel could significantly promote the production of NO in macrophages.(2)In vivo experiment:3 and 10 days after operation,the wound healing rate in the CS-L and CS-D groups was higher than that in the control group(P<0.05).After 21 days,the wound healing rate of the three hydrogel groups was higher than that of the control group.Hematoxylin-eosin staining displayed that a large number of inflammatory cells were infiltrated in the wound tissue of rats in all groups,accompanied by neovessels and fibroblasts 3 days after operation.10 days after operation,there was still more inflammatory cell infiltration in the wound of the control group,and the inflammation of the other three groups was improved,especially the decrease of inflammatory cells in the CS-D group was more obvious.21 days after operation,the wound epithelium of each group was well repaired,and there was basically no inflammatory cell infiltration in the CS-L and CS-D groups,while there was still a small amount of inflammatory cell infiltration in the control group.Immunofluorescence staining revealed that the number of M2-type macrophages in the CS-D group was higher than that in the other three groups(P<0.05).(3)The results conclude that chitosan hydrogels modified by different configurations of arginine can promote wound healing through different mechanisms.

Objective:To evaluate the effect of electroacupuncture (EA) on programmed cell death ligand-1 (PD-L1)/programmed cell death receptor-1 (PD-1)/Src-homology region two domain-containing phosphatase-1 (SHP-1) signaling pathway in the dorsal root ganglia (DRG) of rats with acute postoperative pain.Methods:Thirty-nine SPF male Sprague-Dawley rats, weighing 220-250 g, aged 6-8 weeks, were divided into 3 groups ( n=13 each) using a random number table method: control group (group C), abdominal surgery group (group S) and abdominal surgery+ EA group (group S+ EA). Group S and group S+ EA underwent abdominal surgery under isoflurane anesthesia. In S+ EA group, both Zusanli and Sanyinjiao acupoints were selected and stimulated for 30 min with a frequency of 10 Hz continuous wave and a current of 1 mA starting from the end of operation and 2 h after operation. Seven rats were selected from each group for measurement of the mechanical paw withdrawal threshold (MWT), abdominal contraction threshold (ACT), thermal paw withdrawal latency (TWL) and cold paw withdrawal latency (CWL) at 1 day before developing the model (T 0) and 3, 5, 7, 9, 11 and 24 h after developing the model (T 1-6). Six rats in each group were sacrificed at 5 h after the model was prepared, and T 12-L 4 DRG was removed for determination of the expression of interleukin-6 (IL-6), PD-L1, PD-1 and SHP-1 (by Western blot) and expression of PD-L1 in various neurons (by immunofluorescence). Results:There was no significant difference in TWL and CWL between the three groups at different time points ( P>0.05). Compared with group C, MWT at T 1-5 and ACT at T 1-6 were significantly decreased, and the expression of IL-6 in DRG was up-regulated, and the expression of PD-L1, PD-1 and SHP-1 in DRG was down-regulated in group S ( P<0.05). Compared with group S, MWT and ACT were significantly increased at T 1, 2, the expression of IL-6 in DRG was down-regulated, and the expression of PD-L1, PD-1 and SHP-1 in DRG was up-regulated in group S+ EA ( P<0.05). The results of immunofluorescence showed that PD-L1 was expressed on both small diameter neurons (CGRP + neurons and IB4 + neurons) and large diameter neurons (NF200 + neurons) and mainly on CGRP + neurons and IB4 + neurons. There was no significant difference in the expression of PD-L1 in various DRG neurons among the three groups ( P>0.05). Conclusions:The mechanism by which EA relieves acute postoperative pain may be related to activation of the PD-L1/PD-1-SHP-1 signaling pathway in the DRG of rats.

OBJECTIVETo study the influence of heat stimulation on expression of coxsackie-adenovirus receptor (CAR) in keratinocytes (KCs) of mouse skin and the effect of CAR on production of cell growth factors by dendritic epidermal T lymphocytes (DETCs).

METHODS(1) Twenty BALB/c mice were divided into heat stimulation group (HS) and control group (C) according to the random number table, with 10 mice in each group. Mice in group HS were inflicted with scald milder than superficial-thickness by dressing wet hot gauze, which had been soaked in 100°C hot water for 3 min, in the hair removed area on the back for 1 to 3 s, while mice in group C were sham injured by dressing a wet gauze which had been soaked in water of room temperature for 3 min in the hair removed area on the back for 1 to 3 s. Square full-thickness skin specimens measuring 2.0 cm × 2.0 cm in size were obtained from the center of the bare skin. The expression of CAR in skin tissue sections were detected by immunohistochemistry staining. The mRNA and protein expression levels of CAR in skin tissue sections were respectively determined by real-time fluorescent quantitation RT-PCR and Western blotting. (2) KCs were isolated and cultured from full-thickness skin obtained from the trunk of 2 fetal BALB/c mice, and they were divided into 2 groups according to the random number table, with 5 wells in each group. The cells in group HS and group C were respectively cultured in 42°C and 37°C, 5% CO2 incubator for 1 h, and then all the cells were cultured in 37 °, 5% CO2 incubator for 6 h. The apoptosis of the cells and their expression of CAR were detected by flow cytometer. (3) Five BALB/c mice were sacrificed, and full-thickness skin was obtained from the trunk. The DETCs were divided into 7 groups according to the random number table after being isolated and purified from the skin specimens. Cells in group C were cultured without any stimulation, and cells in the 0.5, 1.0, 2.0, 4.0, 8.0, and 16.0 mg/L CAR groups were respectively cultured with corresponding concentration of recombinant mice CAR nutrient solutions, with 5 wells in each group. The contents of insulin-like growth factor I (IGF-I) and keratinocyte growth factor (KGF) were determined with ELISA. Data were processed with independent samples t test and one-way analysis of variance.

RESULTS(1) The immunohistochemistry staining showed that there was mild positive staining in the skin tissue sections of mice in group C, while the positive staining was more obvious in group HS. The positive staining was mainly located in KCs, hair follicles, and sweat gland epithelial cells, while no positive staining was observed in fibroblasts. The mRNA expression levels of CAR in skin tissue sections in group C and group HS were respectively 0.157 ± 0.027 and 0.773 ± 0.029. There was statistically significant difference between them (t = 3.052, P < 0.01). The protein expression levels of CAR in skin tissue sections in group C and group HS were respectively 0.23 ± 0.09 and 0.89 ± 0.14. There was statistically significant difference between them (t = 2.556, P < 0.05). (2) The apoptosis rates of KCs in group C and group HS were respectively (5.7 ± 1.3)% and (7.4 ± 1.7)% (t = 0.464, P > 0.05). The expression rates of CAR in KCs in group C and group HS were respectively (48 ± 6)% and (80 ± 8)%. There was statistically significant difference between them (t = 2.585, P < 0.05). (3) The contents of IGF-Iin culture supernatants in group C and 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mg/L CAR groups were respectively (23.1 ± 1.8), (22.5 ± 2.1), (31.2 ± 2.5), (39.7 ± 2.3), (61.8 ± 3.5), (45.1 ± 2.8), and (29.0 ± 2.0) µg/L. There was statistically significant difference among 7 groups (F = 3.414, P < 0.05). The contents of KGF in culture supernatants in group C and 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mg/L CAR groups were respectively (131 ± 9), (217 ± 12), (355 ± 21), (563 ± 21), (535 ± 34), (292 ± 20), and (245 ± 10) ng/L. There was statistically significant difference among 7 groups (F = 5.063, P < 0.01).

CONCLUSIONSThe expression of CAR in KCs would rise after HS. The optimum CAR concentration to increase IGF-I and KGF production in DETCs is low.

Animals ; Burns ; metabolism ; Cells, Cultured ; Coxsackie and Adenovirus Receptor-Like Membrane Protein ; metabolism ; Female ; Fibroblast Growth Factor 7 ; metabolism ; Hot Temperature ; Insulin-Like Growth Factor I ; metabolism ; Keratinocytes ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Skin ; cytology ; T-Lymphocytes ; metabolism

Objective : To compare and analyze the effects of thermal etching on the shear strength of zirconia substrates and decorative ceramics. Methods: A total of 20 specimens made with zirconia ceramics were randomly divided into an observation group and a control group with 10 cases in each group. The control group was treated with sandblasting, while the observation group was treated with sandblasting and thermal etching. The surface characteristics were examined by scanning electron microscope (SEM) and phase analysis of X-ray diffraction (XRD), and the shear strength was tested using a universal testing machine. The characteristics of surface destruction were examined by SEM. Results : SEM showed that the peak structure was observed in both groups. The observation group exhibited deep fissures, and the control group exhibited small fissures. The diffraction peaks of the two groups are similar. The T (101) peak is the main peak, and both groups exhibit an M (111) peak. However, the peak intensity is relatively small. The relative levels of monoclinic zirconia were 15.16% in the observation group and 16.22% in the control group. The shear bond strength of the observation group was 24.74 ± 3.02 MPa, which was significantly higher than that of the control group at 21.09 ± 2.58 MPa. The difference was statistically significant (t=2.599, P=0.021). In the control group, the porcelain residue on the zirconia surface was minimal at low magnification, and the zirconia substrate was obviously exposed. The zirconia surface was similar to cristae obliqua at high magnification, and the porcelain exhibited a scattered distribution. In the observation group, a large amount of residual veneer porcelain remained on the zirconia surface at low magnification, but considerable porcelain was observed at high magnification. Conclusion Thermal etching and sandblasting treatment can improve the shear strength of zirconia substrate.

Objective:To comprehensively evaluated the quality of Sargentodoxae Caulis from different habitats with a combination of indexes and characteristic chromatogram method from Chinese Pharmcopoeia (Edition 2020). Methods:The contents of water content, total ash, ethanolic extract, sulfur dioxide residue, heavy metals and harmful elements, total phenols, chlorogenic acid, salidroside and characteristic chromatogram of 17 batches of Sargentodoxae Caulis were determined. The quality of Sargentodoxae Caulis was comprehensively evaluated by combining chemical pattern recognition method. Results:The water content, total ash content, extracts, and content determination of 17 batches of Sargentodoxae Caulis from different habitats complyed with the provisions of the Chinese Pharmcopoeia (Edition 2020). There were differences in the contents of extracts, chlorogenic acid, and salidroside, among which the content of Anhui origin was higher. A total of 8 common peaks were identified from the 17 batches samples. Conclusion:Comprehensive evaluation of multiple indicators can demonstrate the quality of Sargentodoxae Caulis more correctly, and shows that the quality of Sargentodoxae Caulis from different habitats is different. The quality of Sargentodoxae Caulis from Anhui is better than that from other habitats.