Objective:To explore leuncine-rich repeats and immunoglobulin-like domains protein 1 (LRIG1) expression in oral verrucous carcinoma(OVC) and its possible mechanism of tumor suppression.Methods:Paraffin specimens of OVC (n =15) and oral squamous cell carcinoma(OSCC,n =30) and corresponding adjacent normal tissues,and 15 normal mucosa tissues(NM) as control from injured patients were collected.The expression levels of LRIG1 and Bcl-2 were examined using immunohistochemical SP method.The correlation between the expression levels of LRIG1 and Bcl-2 was determined using two tailed Pearson's correlation.Results:LRIG1 was significantly lower in OVC tissue than that in NM tissue,but higher than in OSCC tissue.The expression levels of Bcl-2 in OVC were significantly higher than that in NM,but lower than in OSCC.The expression level of LRIG1 was negatively correlated with expression level of Bcl-2 in NM,OVC and OSCC tissue.Conclusion:LRIG1 may inhibit the expression of Bcl-2 and the development of OVC.

Objective:To investigate the mechanism of Buguozhi Decoction in improving learning-memory of vascular dementia model rats.Methods:The rat models with VaD were established by bilateral occlusion of both common carotid arteries(2 一V0).The gene expressions of ER-?,NR2B were evaluated by RT-PCR and immunohistochemistry.Results:Buguozhi Decoction can significantly improve mRNA and protien pruducts of ER-?,NR2B gene in hippocampus of the vascular dementia model rats,compared with the model group(P

Purpose To explore the value of spectral CT imaging in differentiating pancreatic carcinoma and mass-forming pancreatitis, and to improve the preoperative diagnosis accuracy. Materials and Methods The dual-phase spectral CT imaging data in patients with pathology-proven pancreatic carcinoma (n=62) and mass-forming pancreatitis (n=30) were retrospectively analyzed. At 40-140 keV (with 10 keV increment), the mean CT values, energy spectrum curves and iodine concentrations were measured and compared between pancreatic carcinoma and mass-forming pancreatitis in arterial phase and portal venous phase. Results The mean CT values of mass-forming pancreatitis at 40-140 keV were higher than pancreatic carcinoma in both arterial phase and portal venous phase with significant difference (t= -7.349- -4.810, P<0.05; t= -6.760- -5.496, P<0.05). The energy spectrum curves were significantly different between pancreatic carcinoma and mass-forming focal pancreatitis, especially at 40-70 keV. The iodine concentrations of pancreatic carcinoma were lower than mass-forming pancreatitis in arterial phase and portal venous phase with significant difference (t=-28.577, P<0.05; t= -28.451, P<0.05). Conclusion Pancreatic carcinoma and mass-forming pancreatitis can be differentiated by measuring the CT value at different energies, energy spectrum curves and iodine concentrations.

To investigate the effects of adipose-derived mesenchymal stem cells (AMSCs) from type 2 diabetes mellitus patients on wound healing of pressure ulcers in mice.Methods: (1) In September 2016, the subcutaneous adipose tissue of a 60-year-old woman with type 2 diabetes mellitus was harvested, and then AMSCs were extracted by collagenase digestion and cultured. The third passage of cells were used for subsequent experiments. The morphology of cells was observed, and their osteogenic, chondrogenic, and adipogenic differentiation abilities were identified. The expressions of cell surface markers CD90, CD105, CD73, and CD34 were detected by flow cytometer (n=3). (2) Sixteen female C57BL/6 wild-type mice aged 6-8 weeks were selected, and one pressure ulcer wound was created on each side of the spine of each mouse by pressing the skin with two magnets. The two wounds of each mouse were paired and divided into diabetic AMSCs group and negative control group, injected with 100 μL phosphate buffer solution (PBS) containing green fluorescent protein-labeled AMSCs (1×10⁶ cells) and 100 μL PBS, respectively. The wound healing status of the two groups within post injection day (PID) 21 was observed, and their wound healing rates on PID 5, 13, and 17 were calculated. Three mice were sacrificed on PID 11 and 21, respectively, and tissue of three wounds was harvested from each group. The skin structure was observed by hematoxylin-eosin staining, the collagen deposition was evaluated by Masson staining, and the positive expression of CD31, i. e., the number of new blood vessels was counted by immunohistochemistry. Wound tissue samples of two groups prepared on PID 21 as above-mentioned were harvested, and the positive cell rate of S100, representing the regeneration of Schwann cells, was detected by immunohistochemistry. Wound tissue samples of diabetic AMSCs group prepared on PID 11 as above-mentioned were harvested, and the colonization of AMSCs was observed by fluorescence tracer method. Data were processed with paired t test and Bonferroni correction. Results: (1) The third passage of cells isolated and cultured from the subcutaneous adipose tissue of a type 2 diabetes mellitus patient grew adherently to the wall in a long spindle and vortex-like manner. After induction, the cells showed osteogenic, chondrogenic, and lipogenic differentiation abilities. The positive expression rates of CD90, CD105, and CD73 on the cell surface were higher than 90.00%, and the expression rate of CD34 was 0.46%. The cells were identified as AMSCs. (2) The mice wounds of diabetic AMSCs group healed quickly, and all the wounds healed completely on PID 17, while the mice wounds in negative control group were not completely closed at this time, and there was still scab on the surface. On PID 5, 13, and 17, the healing rates of mice wounds of diabetic AMSCs group were (35.6±6.5)%, (87.1±2.5)%, and 100.0%, respectively, significantly higher than (19.8±7.2)%, (66.2±5.2)%, and (86.9±5.3)% of negative control group (t=6.49, 14.31, 9.73, P<0.05). Compared with that of negative control group, the inflammatory cell infiltration was reduced in mice wounds tissue of diabetic AMSCs group on PID 11, and thicker epidermis and dermis as well as regenerated skin appendages were observed on PID 21. On PID 11 and 21, the collagen percentages of mice wounds tissue in diabetic AMSCs group was (48.3±1.3)% and (54.1±1.7)%, respectively, significantly higher than (41.4±1.7)% and (50.3±1.2)% of negative control group (t=6.98, 3.99, P<0.01). On PID 11 and 21, the numbers of new blood vessels in mice wounds tissue of diabetic AMSCs group were 17.2±1.3 and 18.0±2.1, respectively, significantly more than 8.0±1.4 and 14.0±1.5 of negative control group (t=10.69, 3.38, P<0.01). On PID 21, the S100 positive cell percentage in mice wounds tissue of diabetic AMSCs group was (1.76±0.12)%, significantly higher than (0.55±0.03)% of negative control group (t=21.68, P<0.001). On PID 11, the colonization of AMSCs in mice wounds tissue of diabetic AMSCs group was observed. Conclusions Transplantation of AMSCs from type 2 diabetic mellitus patients can accelerate wound healing of pressure ulcers in mice by promoting angiogenesis, collagen deposition, and Schwann cell regeneration.

Objective: To explore the effects of local transplantation of autologous adipose-derived stromal vascular fraction (SVF) on the hyperplastic scar (HS) formation in rabbit ears and the mechanism. Methods: Twenty-four New Zealand white rabbits were used to reproduce HSs by making four full-thickness skin defect wounds with a diameter of 1 cm on the ventral surface of left ear of each rabbit. Wound epithelization and local-tissue proliferation were observed, and wound healing (complete epithelization) time and formation time of HS were recorded. The 24 rabbits were divided into SVF group, pure DMEM group, and pure HS group according to the random number table, with 8 rabbits and 32 wounds in each group. On post injury day (PID) 25 (after the complete epithelization of wounds), 0.2 mL of low glucose DMEM medium containing CM-Dil labeled autologous SVF was injected into HSs of rabbits in SVF group, while the same amount of low glucose DMEM medium was injected into HSs of rabbits in pure DMEM group. The frequency of injection was once every 5 days, totally for 3 times. HSs of rabbits in pure HS group did not receive any treatment. On PID 40, HSs of rabbits′ ears in each group were harvested, then the histological form was observed by hematoxylin and eosin staining, the arrangement of collagen in HS was observed by Van Gieson staining, the distribution of CM-Dil-labeled SVF in the HS was observed with fluorescence microscope, and the mRNA expression and the protein expression of transforming growth factor β₁ (TGF-β₁), Smad3, and Smad7 in HS were determined by real-time fluorescent quantitative reverse transcription-polymerase chain reaction and Western blotting, respectively. Data were processed with one-way analysis of variance and Tukey test. Results: (1) Complete epithelization time of wounds of rabbits′ ears was (20.0±2.0) d post injury, and HSs were formed on PID 25. On PID 40, HSs of rabbits′ ears in pure DMEM group and pure HS group were still in hyperplasia, while those in SVF group became smaller, flat, soft, and light colored. (2) On PID 40, compared with those in pure DMEM group and pure HS group, the number of epithelium foot like structures was more and the amount of inflammatory cells was less. The collagen of HSs of rabbits′ ears in SVF group was arranged more regularly with broader gap between collagens. (3) On PID 40, CM-Dil-labeled SVF could still be observed in the HSs of rabbits′ ears in SVF group. (4) On PID 40, compared with those in pure DMEM group and pure HS group, the mRNA expressions of TGF-β₁ and Smad3 in the HSs of rabbits′ ears in SVF group were significantly down-regulated (P<0.05), while the mRNA expression of Smad7 was significantly up-regulated (P<0.05). There were no significant differences in the mRNA expressions of TGF-β₁, Smad3, and Smad7 in the HSs of rabbits′ ears between pure DMEM group and pure HS group (P>0.05). (5) On PID 40, compared with those in pure DMEM group (0.74±0.03, 0.73±0.10, 0.54±0.09) and pure HS group (0.72±0.08, 0.71±0.12, 0.53±0.06), the protein expressions of TGF-β₁ and Smad3 in the HSs of rabbits′ ears in SVF group (0.57±0.06, 0.42±0.09) were significantly down-regulated (P<0.05), while the protein expression of Smad7 (0.71±0.05) was significantly up-regulated (P<0.05). The protein expressions of TGF-β₁, Smad3, and Smad7 in the HSs of rabbits′ ears in pure DMEM group and pure HS group were close (P>0.05). Conclusions Autologous SVF transplantation can inhibit the formation of HS in the early stage of scar formation of rabbit, the mechanism may be related to the TGF-β₁/Smad signaling pathway.

Objective:To investigate the effects of combination treatment of photodynamic therapy (PDT) based on photosensitizer chlorin e6 (Ce6) and antibiotic agent tinidazole (TNZ) against periodontitis both in vitro and in vivo. Methods:The Sprague-Dewley (SD) rat periodontitis model was constructed using the method of orthodontic wire ligation. After successful modeling, SD rats were randomly divided into the following 6 groups (3 rats in each group): positive control (Ctrl+), Ce6, TNZ, a mixture of Ce6 and TNZ (Ce6/TNZ), Ce6 with laser irradiation (Ce6+L), a mixture of Ce6 and TNZ with laser irradiation (Ce6/TNZ+L). Methyl thiazolyl tetrazolium (MTT) assay was used to assess the cytotoxic activities of Ce6 (concentration range: 0-20 mg/L), TNZ (concentration range: 0-16.6 mg/L) and their mixture (Ce6/TNZ) in mouse fibroblast L929 cells. Fluorescence probe method was applied to measure the production of reactive oxygen species in the dental plaque biofilms after various treatments with and without 5-minute laser irradiation at 635 nm at a power density of 0.5 W/cm 2 (Ce6+L and Ce6/TNZ+L groups), thus to evaluate the PDT performances. Cell counting kit-8 (CCK-8) and live/dead staining were used to assess the antibacterial activity in each of the groups and the combination index (CI) of PDT combined with TNZ was calculated subsequently. Flow cytometry was utilized to detect the apoptosis-inducing effects of these treatments in macrophage RAW264.7 cells after processing with the apoptosis detection kit. The inhibitory effects of various treatments on the absorption of alveolar bone of SD rats were further evaluated in the periodontitis rats by using the micro-CT. Results:The survival rates of L929 cells in the preset concentration range were all above 90% in Ce6, TNZ and Ce6/TNZ groups. Upon laser irradiation, the plaque biofilms in Ce6 and Ce6/TNZ groups showed significant green fluorescence, indicating that large amounts of reactive oxygen species were triggered and generated significantly in the dental plaque biofilms. However, the survival rates of dental plaque microorganisms in 5 Ce6/TNZ concentrations were (85.4±5.5)%, (76.0±8.9)%, (61.7±0.6)%, (56.3±2.6)% and (43.5±0.6)% respectively, which were significantly lower than that in Ce6 only and TNZ only groups ( P<0.01). The CI levle of each drug concentration group was less than 1.0, which showed a significant synergistic antibacterial efficiency. Stronger apoptotic activities were observed in Ce6+L and Ce6/TNZ+L groups compared with those in Ce6 only and Ce6/TNZ only groups ( P<0.01). In periodontitis rats, Ce6/TNZ combined laser irradiation could effectively inhibit the absorption of alveolar bone. The alveolar bone volume and the ratio of bone volume and tissue volume were (1.49±0.07) mm 3 and (47.08±0.71)%, respectively. The distances between cementoenamel junction to alveolar bone crest on buccal and palatal sites decreased to (2.13±0.07) mm and (1.94±0.10) mm respectively, showing a high inhibition efficiency. Conclusions:Ce6-mediated PDT combined with TNZ possessed notable synergistic effects against periodontitis, reflecting in the efficient antibacterial effect, the apoptosis-inducing action on macrophages, and the inhibitory efficacy on the alveolar bone absorption in vivo.

OBJECTIVETo explore the effect of heat shock protein 90 (HSP90) inhibitor (17-DMAG) and oxaliplatin on the proliferation and invasion of colorectal cancer.

METHODSAfter 17-DMAG, oxaliplatin and half-dose combination of 2 drugs processing colorectal cancer SW480 and HCT116 cell lines, CCK8 assay was applied to detect cell viability. RT-PCR and Western blot were used to detect the expression level of the apoptosis-related molecules. Transwell chemokine axis experiment and Western blot were employed to detect cell invasion ability and the expression level of tumor metastasis-associated protein.

RESULTSThe growth of SW480 and HCT116 cells was inhibited after the administration of 17-DMAG and oxaliplatin(P<0.05) in dose- and time-dependent manner. Processed by 17-DMAG 100 nmol/L, oxaliplatin 50 mg/L and half-dose combination of 2 drugs, transcription level of the apoptosis inhibitory gene (Bcl-2) in SW480 and HCT116 cells was decreased, the level of apoptosis promoting gene (Bax) transcription and protein PARP-1 spliceosome expression was increased, and the above trend was more obvious when using half-dose combination of 2 drugs. Transwell chemokine axis experiments showed the penetrating relative percentage and expression level of MMP9 and integrin β3 decreased, especially for half-dose combination of 2 drugs.

CONCLUSION17-DMAG and oxaliplatin can co-inhibit the proliferation and invasion of colorectal cancer.

Antineoplastic Agents ; Apoptosis ; Benzoquinones ; Cell Proliferation ; Cell Survival ; Colorectal Neoplasms ; HCT116 Cells ; Humans ; Lactams, Macrocyclic ; Neoplasm Invasiveness ; Organoplatinum Compounds

Objective To evaluate the clinical efficacy of single plate,double plates,and intramedullary nail in treating medial cortical comminuted fractures of the medial distal femur.Methods A retrospective case control study was conducted on 63 cases of medial cortical comminuted fractures of the medial distal femur treated from November 2011 through October 2016.According to the type of internal fixation,19 cases of middle and distal shaft fractures were divided into the single plate group (n =11) comprising four males and seven females,aged averagely 57.5 years,and intramedullary nail group (n =8) comprising five males and three females,aged averagely 56.4 years.Forty-four cases of distal femoral fractures were divided into the single plate group (n =14) comprising six males and eight females,aged averagely 57.6 years,intramedullary nail group (n =9) comprising five males and four females,aged averagely 56.6 years,and double plates group (n =21) comprising 14 males and seven females,aged averagely 55.1 years.The rate and period of racture healing,distal femoral valgus resection (DFVR),femoral angle,change of DFVR and femoral angle,and complications were compared within and between groups.Results All the cases were followed up for 6-37 months (mean,13.4 months).Of 19 cases of middle and distal shaft fractures,there were no statistically significant differences in fracture healing rate and period,DFVR after operation and at the last follow-up,and the amount of change of DFVR between the single plate and double plates group (P ＞ 0.05).With respect to postoperative complications of the two groups,no significant difference was found for the rate of nonunion,infection,internal fixation failure,and mal union (P ＞ 0.05).However,the incidence rate of adverse events in single plate group (46％) was significantly higher than that in intramedullary nail group (0) (P ＜0.05).Of the 44 cases of distal femoral fractures,the pairwise comparisons among three groups showed no statistically significant differences in fracture healing rate,healing period,and the post operative femoral angle (P ＞ 0.05).The femoral angle at the last follow-up in intramcdullary nail group was (80.80 ± 2.93) °,significantly lower than (85.28 ± 5.89) ° in single plate group and (83.55 ± 3.51) ° in double plates group (P ＜ 0.05).Within the intramedullary nail group,no significant difference was found between the femoral angle immediately after operation and at the last follow-up (P ＞ 0.05),but significant difference was detected in both single plate group and double plates group (P ＜ 0.05).The amount of change of the femoral angle in intramedullary nail group [(0.25 ± 1.95) °] was significantly lower than (4.03 ± 3.78) ° in single plate group and (2.60 ± 2.24) ° in double plates group (P ＜ 0.05).With respect to post operative complications in the three groups,no significant difference was found for the rate of nonunion,infection,internal fixation failure,and malunion (P ＞ 0.05).The malunion rate in single plate group (36％) was significantly higher than that in double plates group (14％).The incidence rate of adverse events in single plate group (50％) was significantly higher than that in double plates group (14％) (P ＜ 0.05).Conclusions Three internal fixations have similar clinical results in fracture healing rate and period.For medial cortical comminuted middle and distal shaft fractures,and distal femoral fractures without involving articular surface,intramedullary nail has better results than locking plate.For medial cortical comminuted distal femoral fractures involving articular surface,double plates fixation might be a better option.

Objective To explore the effect of heat shock protein 90 (HSP90) inhibitor (17-DMAG) and oxaliplatin on the proliferation and invasion of colorectal cancer. Methods After 17-DMAG, oxaliplatin and half-dose combination of 2 drugs processing colorectal cancer SW480 and HCT116 cell lines, CCK8 assay was applied to detect cell viability. RT-PCR and Western blot were used to detect the expression level of the apoptosis-related molecules. Transwell chemokine axis experiment and Western blot were employed to detect cell invasion ability and the expression level of tumor metastasis-associated protein. Results The growth of SW480 and HCT116 cells was inhibited after the administration of 17-DMAG and oxaliplatin (P<0.05) in dose- and time-dependent manner. Processed by 17-DMAG 100 nmol/L, oxaliplatin 50 mg/L and half-dose combination of 2 drugs, transcription level of the apoptosis inhibitory gene (Bcl-2) in SW480 and HCT116 cells was decreased, the level of apoptosis promoting gene (Bax) transcription and protein PARP-1 spliceosome expression was increased, and the above trend was more obvious when using half-dose combination of 2 drugs. Transwell chemokine axis experiments showed the penetrating relative percentage and expression level of MMP9 and integrin β3 decrease d, especially for half-dose combination of 2 drugs. Conclusion 17-DMAG and oxaliplatin can co-inhibit the proliferation and invasion of colorectal cancer.

Objective:To investigate the clinical effects of modified fascia flap from cutaneous branch of dorsal metacarpal artery in repairing the wound at the proximal and middle finger segments.Methods:From January 2017 to September 2018, 12 patients with wounds at the proximal and middle finger segments were admitted to the Affiliated Hospital of Zunyi Medical University, including 8 males and 4 females, aged 35-70 years. The areas of wounds ranged from 3.4 cm×2.4 cm to 6.5 cm×4.0 cm. The modified fascia flaps from cutaneous branch of dorsal metacarpal artery were resected to repair the wounds, with the size ranging from 3.5 cm×2.5 cm to 6.7 cm×4.1 cm. The flap donor sites of 5 patients were repaired with direct intermittent suture, the flap donor sites of 4 patients were repaired with full-thickness skin grafts from ipsilateral medial forearm, and the flap donor sites of 3 patients were repaired with wrist pedicled flaps. The survival of the flaps was recorded. Healing of donor site and recipient site was followed. The hand functions were evaluated with trial standard for the evaluation of the functions of the upper limbs of the Hand Surgery Society of the Chinese Medical Association.Results:All the flaps survived in 12 cases. During 3 to 12 months of follow-up, the flaps recovered satisfactorily in texture and shape. The donor sites of 11 patients were healed, and the skin graft edge area was partially necrotic in the other patient but healed later after dressing change. The distances of two-point discrimination of the patients ranged from 5.6 to 9.0 mm. Hand functions were evaluated as excellent in 5 cases, good in 4 cases, and fair in 3 cases.Conclusions:Modified fascia flap from cutaneous branch of dorsal metacarpal artery for repairing the wounds at the proximal and middle finger segments has reliable blood supply. The operation is simple and safe with short course of treatment, which is worthy of clinical promotion.