Background Mesenchymal stem cells (MSCs) have been applied in basic and clinical researches of organ transplantation.Our previous study showed that intravenous injection of MSCs prolonged corneal allograft survival in rat.However,the effect of local administration of MSCs on corneal allograft rejection remains unclear.Objective The aim of this study was to investigate the effect of subconjunctival injection of MSCs on corneal allograft rejection in rat model of keratoplasty.Methods MSCs were isolated and cultured from femur and tibia bone marrow of clean Wistar rats,and then the cells were identified by induced differentiation of osteoblast and adipocyte.The third generation of MSCs was used in subsequent experiment.Allogenic penetrating keratoplasty was performed with the bilateral corneas of 20 Wistar rats as donor grafts and the right eyes of 40 Lewis rats as recipients.PBS 0.1 ml containing 2 × 106 MSCs and 0.1 ml PBS only was subconjunctivally injected immediately and postoperative 3 days respectively in randomized two groups,and another 6 normal Lewis rats served as the normal control group.Corneal rejection response was evaluated under the slit lamp after surgery based corneal opacity,edema and neovascularization,and the grafts were scored according to the criteria of Larkin.The corneal samples were extracted from 12 rats of the PBS control group and the MSCs group separately 10 days after surgery.The relative expressions of Th1 cytokines (interferon-γ [IFN-γ] mRNA and interleukin-2 [IL-2] mRNA) and Th2 cytokines (IL-4 mRNA and IL-10 mRNA) were detected by real-time quantitative PCR.Protein levels of IL-4 and IL-10 proteins in the corneas were assayed by ELISA.All experimental protocols involving rats were approved by the laboratory animal care and use committee of the Tianjin Medical University and treated with the ARVO statement for the use of animals in ophthalmic and vision research.Results The cells grew well with the orange stain for alizarin red in differentiated the osteoblasts and red stain for Oil red O in differentiated adipocytes.The survival time of corneal graft in the MSCs group was (11.8±1.6) days,it was significantly longer than (9.6±1.4) days in the PBS control group (P=0.004).The levels of IL-4 mRNA and IL-10 mRNA in the MSCs group were significantly higher than those in the PBS control group (both at P =0.00);while the levels of IFN-γ mRNA and IL-2 mRNA were not significantly different between the groups (both at P＞0.05).The IL-10 protein contents were (22.74 ±7.06),(68.40±12.83) and (215.41 ±44.66)pg/ml in the normal control group,PBS control group and MSCs group,showing significant difference among the three groups (F =55.06,P =0.00) and a significant increase in the MSCs group compared with the PBS control group and the normal control group (both at P ＜ 0.05).Conclusions Subconjunctival injection of MSCs prolongs the survival time of cornea allograft in penetrating keratoplasty probably by modulating the balance between Th1 and Th2 cytokines,especially by up-regulating Th2 cytokines.

OBJECTIVETo explore the clinical efficacy and action mechanism of acupuncture for premature ovarian failure (POF).

METHODSAccording to prospective cohort study design, 30 cases were included. Based on theory of regulating Chong and Conception Vessels as well as soothing the liver and calming the nerves, acupuncture at Guanyuan (CV 4), Dahe (KI 12), Taixi (KI 3), Taichong (LR 3), Baihui (GV 20) was applied, three times per week and 3 months were considered as one session. Totally two sessions were performed. The menstruation condition, estradiol (E2), follicle-stimulating hormone (FSH) and scores of the clinical perimenopausal symptoms were taken as treatment outcomes.

RESULTSThe total effective rate was 86.7% (26/30) and the cured rate was 16.7% (5/30). The recovery rate of menstruation was 16.7% (5/30) and the regain rate of menstruation was 56.7% (17/30). After the treatment, the level of E2 was significantly increased from (45.41 +/- 18. 09) pmol/L to (59.07 +/- 24.21) pmol/L (P < 0.05), and the score of perimenopausal symptoms was obviously reduced from 14.28 +/- 8. 30 to 4.04 +/- 3.28 (P < 0.01). However, no statistical differences could be seen in FSH before and after treatment (P > 0.05).

CONCLUSIONAcupuncture has certain effect on improving menstruation and perimenopausal symptoms in POF patients, which is likely to he related with increasing the level of E2.

Acupuncture Therapy ; Adolescent ; Estradiol ; metabolism ; Female ; Humans ; Luteinizing Hormone ; metabolism ; Primary Ovarian Insufficiency ; metabolism ; therapy ; Prospective Studies ; Young Adult

Objective To investigate the role of expressions of pituitary tumor transforming gene(PTTG) and p27 in glioma,and explore the relationship between the expressions of them and cell cycle regulation.Methods Specimens of 40 patient with gliomas were divided into gradeⅠ:8 cases,gradeⅡ:10 cases,gradeⅢ:13 cases, gradeⅣ:9 cases,PTTGmRNA and p27mRNA were detected by reverse transcription polymerase chain reaction (RT-PCR),and their expressions of PTTG,P27,CDK4 protein were detected by immunostaining assay using strep- tavidin-peroxidase(SP) method.Results The expressions of PTTGmRNA in gradeⅠ～Ⅳwere (0.907?0.065),(1.109?0.083),(1.312?0.089),(1.499?0.215) respectively,there was significant difference among the groups(P

AIM: To investigate the efficacy and safety of needle revision with 5-fluorouracil (5-FU) on the dysfunctional filtering blebs after trabeculectomy and to assess the factors that may impact the success.
METHODS:Eighty- three eyes in 76 patients underwent the needle revision and 5-FU subconjunctive injection for the dysfunctional blebs after trabeculectomy and were followed up for 12mo. The intraocular pressure ( IOP), the number of drugs, corneal endothclium, bleb morphology and complications were observed and recorded.
RESULTS: IOP decreased significantly from 35. 3 ±5. 8mmHg(1kPa = 7. 5mmHg) of pre - needling to 17. 0 ±4. 3mmHg of post - needling ( P < 0. 01 ); the average numbers of medications decreased significantly from 1. 7±0. 9 of pre-needing to 0. 4±0. 7 of post-needing (P<0. 01). At 12mo after needling, the success rate of filtering blebs was 89. 2% and the complete success rate was 69. 9%. The Kaplan - Meier survival analysis estimated mean survival period was 11. 0mo (95% CI: 10. 3 - 11. 6). Statistically, there were no significant difference on needling effect with reference to the types of glaucoma, the use of mitomycin C ( MMC ) during the previous filtration surgery, the ages of patients, the intervals of needling operation from previous trabeculectomy, while there were significant difference on needling effect with reference to bleb appearance before needling, and the mean number of needling in patients that had surgery within 3mo were less than those who had surgery for more than 3mo.
CONCLUSION: The needle revision combined with 5-FU is a safe, effective and simple method. Dysfunctional blebs should be treated early after trabeculectomy.

The aim of this paper was to investigate the effect of terpene penetration enhancers on membrane fluidity and membrane potential using HaCaT keratinocytes, and study the potential mechanisms of these terpene compounds using as natural transdermal penetration enhancer. Six terpene compounds, namely menthol, limonene, 1,8-cineole, menthone, terpinen-4-ol and pulegone, were chosen in this study on account of their good penetration-enhancement activities. The cytotoxicity of these terpene compounds was measured using an MTT assay. The fluorescence recovery after photobleaching (FRAP) technique was employed to measure the change of membrane fluidity of HaCaT cells. The flow cytometer was used to study the alteration of membrane fluidity of HaCaT cells, and investigate the effect of terpene compounds on intracellular Ca2+. It was found that 6 terpene compounds possessed low cytotoxicity in comparison to the well-established and standard penetration enhancer azone. Those terpene compounds could significantly enhance HaCaT cells membrane fluidity and decrease HaCaT cells membrane potentials. Meanwhile, after treated with various terpene compounds, the Ca2(+)-ATPase activity and intracellular Ca2+ of HaCaT cells was decreased significantly. Terpene penetration enhancers perhaps changed the membrane fluidity and potentials of HaCaT cells by altering the Ca2+ balance of the cell inside and outside, resulting in the low skin permeability to increase the drug transdermal absorption.
Cell Line ; Drugs, Chinese Herbal ; pharmacokinetics ; Humans ; Keratinocytes ; drug effects ; metabolism ; Membrane Fluidity ; drug effects ; Skin Absorption ; drug effects ; Terpenes ; pharmacokinetics

Objective: To evaluate the accuracy of automated classification of ICD-O-3 morphology code from pathology reports by text-mining and support vector machine ( SVM ) , in order to provide basis for automated tumor coding in Chinese. Methods: The tumor report cards of Zhejiang residents from 2017 to 2019 were collected from Chronic Disease Surveillance Information Management System of Zhejiang Province. According to ICD-O-3, the keywords of the pathology reports were extracted, and SVM was used for automatic classification. The classification results were compared with those of 16 professionals with more than two years of experience in tumor coding, and the accuracy rate, recall rate and F-score were calculated for effect evaluation. Results: Totally 83 082 cases from 2017 to 2019 were included and were categorized into 17 morphological classifications, with 52 877 ( 63.65% ) cases of adenocarcinoma, squamous carcinoma and transitional cell carcinoma. A total of 1 090 keywords were enrolled into main corpus. The total F-score, accuracy rate and recall rate are 85.69, 77.20% and 96.27%, respectively. Conclusion Text-mining combined with SVM can improve the efficiency of ICD-O-3 morphology coding; however, the accuracy needs to be further improved.

Objective To investigate the effect of cnrcumin on the expression of pituitary tumor transforming gene (PTTG) in glioma C6 cell line. Methods Glioma C6 cells were allocated into the control group and 3 cttrcnmin treatment groups with curcumin treatment at 10, 20 and 30 μmol/L for 24 h. After the treatments, PTTG mRNA and protein expressions in the cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively. Results The expression levels of PTTG mRNA in the 4 groups, showed significant differences between any two groups (P<0.01). Significant differences were also found in PTTG protein expressions between the 4 groups of C6 cells after the treatment (P<0.01). Conclusions Cureumin can down-regulate PTTG expression at both the mRNA and protein levels in glioma C6 cells in a dose-dependent manner.

Objective To investigate the chemotherapeutic sensitivity of glioma cells influenced by RNA interference of pituitary tumor transforming gene (PTTG). Methods The glioma U251 cell line was divided into normal control group, TMZ treatment group, PTTG shRNA infection group and PTTG shRNA combined with TMZ treatment group. Cells in the later 2 groups were treated with PTTG shRNA and PTTG shRNA combined with TMZ. MTT assay and flow cytometry were employed to detect the cell proliferation and apoptosis of U251 cell line, respectively. Results The outcome of MTT assay showed that the growth and survive abilities were influenced after treating with PTTG shRNA and PTTG shRNA combined with TMZ; the optical density (OD) value of the control group, TMZ group,PTTG shRNA treatment group and PTTG shRNA combined with TMZ treatment group were (0.85±0.07),(0.58±0.06), (0.55±0.07) and (0.41 ±0.05), respectively. The inhibitory rate of cell proliferation in the TMZ treatment group, PTTG shRNA treatment group and PTTG shRNA combined with TMZ treatment group were (31.56±5.51 )%, (35.53±4.60)%, (51.49±6.74)%, respectively; statistical significance between control group and both PTTG shRNA group and TMZ group was noted (P＜0.05); and statistical significance between group PTTG shRNA combined with TMZ and both PTTG shRNA group and TMZ group was noted too. The apoptosis rate in the control group, TMZ group, PTTG shRNA group and PTTG shRNA combined with TMZ group were (6.29±0.78)%, (33.63±4.88)%, (39.61 ±4.95)% and (66.23±7.60)%, respectively, 48 h after the treatment; significant differences were found between the control group and both PTTG shRNA group and TMZ group (P＜0.05); and statistical increased apoptosis rate in the PTTG shRNA combined with TMZ group was noted as compared that in the PTTG shRNA group and TMZ group (P＜0.05). Conclusion PTTG RNA interference, by inhibiting the cell proliferation and inducing the apoptosis of glioma cells, up-regulates the chemotherapeutic sensitivity and improve the effectiveness of chemotherapy.

OBJECTIVETo investigate the roles of surviving and caspase-3 in the development of oral cancer.

METHODSArchival tissue sections of 17 oral squamous cell carcinoma (OSCC), 28 oral leukoplakia with dysplasia, 10 normal oral mucosa were obtained from Capital Medical University School of Stomatology for immunohistochemical staining of markers of survivin and caspase-3. The cell apoptosis was detected with terminal deoxynucleotidyl transferase-mediated nucleotide shift enzyme (TdT) mediated d-UTP end labeling (TUNEL). Positively stained cells were counted and analyzed statistically to determine potential relationship between survivin, caspase-3 and cell apoptosis.

RESULTSThe expression of survivin was faint or negative in normal epithelial cells. The average positive rate of survivin was (1.05 ± 1.21)% in control group and (21.89 ± 10.45)% in OSCC. Caspase-3 was expressed in all the normal mucosa,but it obviously down-regulated in dysplasia and OSCC. The apoptosis index (AI) decreased from (0.89 ± 0.46)% in normal mucosa to (0.21 ± 0.12)% in OSCC.

CONCLUSIONSBoth survivin and caspase-3 are associated with carcinogenesis of the oral mucosa. Survivin may restrain cell apoptosis by inhibiting caspase-3.

Apoptosis ; Carcinoma, Squamous Cell ; enzymology ; metabolism ; pathology ; Caspase 3 ; metabolism ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Leukoplakia, Oral ; enzymology ; metabolism ; pathology ; Mouth Mucosa ; enzymology ; metabolism ; Mouth Neoplasms ; enzymology ; metabolism ; pathology ; Precancerous Conditions ; enzymology ; metabolism ; pathology

Objective To compare the spiral CT and pathological findings of sub-centimetre pulmonary ground glass nodules(GGNs), and to improve the aaccuracy in early diagnosis.Methods The CT findings and pathologic features of 30 patients with sub-centimetre pulmo-nary GGNs in our hospital from May 2013 to June 2016 were reviewed retrospectively.The patients were divided into 3 groups according to their pathological classification,namely the benign group,the preinvasive group and the invasive group.The shape,size,margin,vacuole sign, pleural indentation,the relations with bronchi and blood vessels of the pulmonary GGNs were analyzed.Results There was no significant difference in shape,maximum diameter and margin between the benign group and the other two groups(P>0.05).Adjacent vessels changes of the lesion showed significant differences between benign group and preinvasive or invasive groups(P<0.05).And there were significant differences in size,pleural indentation,vacuole sign and adjacent vessels changes between the preinvasive group and the invasive group(P<0.05).Conclusion The comprehensive analysis of lesions size,vacuole sign,pleural indentation and adjacent vessels changes can be helpful to improve the aaccuracy of differential diagnosis on sub-centimetre pulmonary ground glass nodules.