Objective To study cervical human papillomavirus (HPV) infection and genotype distribution characteristics in patients with vaginal disease.Methods The PCR dot blot was used to carry out HPV detection of cervical exfoliated cells in 816 women with cervical lesions in the hospital, and 23 kinds of HPV genotypes were identified.Results Among the 816 cases receiving HPV type detection, HPV of 532 cases (65.20%) was positive, including 371 cases (69.74%) with low-risk HPV (HR-HPV) infection, 161 cases (30.26%) with low-risk HPV infection (LR-HPV) (P<0.05).The HPV infection rate below 45 years old was relatively higher.The HPV infection rate in patients with cervical cancer (97.10%) was higher than that in patients with cervical intraepithelial neoplasia (CIN) II -III (80.54%), patients with CIN Ⅰ(41.55%) and patients with cervicitis (17.20%) (P<0.05).The top five types of HPV infection were HPV16, 58, 18, 11 and 56 type, respectively.The single infection rate (79.70%) was higher than multiple infection rate (20.30%) (P<0.05); HR-HPV infection was the main infection in single infection, and double infection was the main infection in multiple infection.Conclusion Below 45 years old women are the high-risk groups of HPV infection.16, 58, 18, 11 and 56 are the main types of HPV infection.The infection rates of patients with cervical cancer and precancerous lesions are relatively higher.Paying attention to the screening of HPV infection has positive significance in the prevention and treatment of cervical cancer.

Objective: To observe the relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia and evaluate the safety preliminarily.Methods: The relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia were studied by the cough model caused by the irritation of ammonia water and the phenol red output of trachea in mice.The acute toxicity test and maximum tolerance test were carried out to evaluate the safety.Results: The total alkaloid in Atalantia buxifolia at low dose could obviously prolong cough incubation period and decrease cough times in mice, and that at high dose could significantly increase the secretion of phenol red in respiratory tract, and compared with those in the blank group, the differences were statistically significant (P<0.05).In the acute toxicity test, no death showed after the administration with maximum tolerance dosage, and the rate of weight growth had no difference between the blank group and the model group (P>0.05).Conclusion: The relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia are notable in the cough model caused by the irritation of ammonia water and the phenol red output of trachea in mice.The maximum tolerable dose test shows the total alkaloid in Atalantia buxifolia is safe.

Objective: To preliminarily investigate the influence of recombinant human growth hormone (rhGH) on the immune function of younger children with severe burn injuries. Methods: A total of 30 younger children with severe burn injuries, conforming to the study criteria, were admitted to our hospital from July 2016 to July 2018. They were enrolled in the prospective, randomized, double-blinded, controlled trial and divided into group rhGH [n=15, 10 boys and 5 girls, aged (22±10) months] and control group [n=15, 8 boys and 7 girls, aged (21±7) months] according to the random number table. The patients in control group received anti-shock, anti-infection, and wound caring therapies, etc. On the basis of above-mentioned treatment, the patients in group rhGH were subcutaneously injected with rhGH once every night before bedding, with a dosage of 0.2 IU·kg^-1·d^-1, from the 3rd day post injury for 7 consecutive days. Before and on the 3rd and 7th day of rhGH treatments, the fasting peripheral venous blood was collected from patients in both groups. Blood glucose level was detected by glucometer. Percentages of CD4⁺ T lymphocytes, CD8⁺ T lymphocytes, CD3⁺ T lymphocytes, CD19⁺ B lymphocytes, and ratio of CD4⁺ T lymphocytes to CD8⁺ T lymphocytes were determined by flow cytometer. Mass concentration of serum immune globulin (Ig) A, IgG, and complement C3 were detected by enzyme-linked immunosorbent assay. Data were processed with Fisher′s exact probability test, independent sample t test, analysis of variance for repeated measurement and Bonferroni correction, and Mann-Whitney U test. Results: (1) The blood glucose levels of children in the two groups were similar before and on the 3rd and 7th day of rhGH treatment (t=0.474, 1.652, 1.997, P>0.05). The glucose levels of children in group rhGH on the 3rd and 7th day of rhGH treatment [(6.9±1.0) and (7.7±1.1) mmol/L] were significantly higher than (5.9±0.9) mmol/L before rhGH treatment (P<0.05). The glucose level of children in control group on the 7th day of rhGH treatment was significantly higher than that before rhGH treatment (P<0.05). (2) The percentages of CD4⁺ T lymphocytes of children in group rhGH before rhGH treatment and on the 7th day of rhGH treatment were (35.1±2.0)% and (38.5±2.2)%, which were close to (36.2±2.0)% and (33.6±2.2)% in control group, respectively (t=0.371, 1.553, P>0.05). The percentages of CD4⁺ T lymphocytes of children in group rhGH on the 7th day of rhGH treatment[(44.7±2.2)%] was significantly higher than (36.5±2.2)% in control group (t=2.624, P<0.05). The percentage of CD4⁺ T lymphocytes of children in group rhGH on the 7th day of rhGH treatment was significantly higher than that before rhGH treatment (P<0.05). The percentages of CD4⁺ T lymphocytes of children in control group on the 3rd and 7th day of rhGH treatment were both close to the percentage before rhGH treatment (P>0.05). (3) The percentage of CD8⁺ T lymphocytes of children in group rhGH on the 3rd day of rhGH treatment was significantly lower than that in control group (t=2.107, P<0.05). (4) The ratio of CD4⁺ T lymphocytes to CD8⁺ T lymphocytes of children in group rhGH on the 7th day of rhGH treatment (2.36±0.20) was significantly higher than 1.72±0.20 in control group (t=2.285, P<0.05). The ratio of CD4⁺ T lymphocytes to CD8⁺ T lymphocytes of children in group rhGH on the 7th day of rhGH treatment was significantly higher than 2.04±0.19 before rhGH treatment (P<0.05). (5) The percentages of CD3⁺ T lymphocytes and CD19⁺ B lymphocytes of children in the two groups were similar before and on the 3rd and 7th day of rhGH treatment (t=1.913, 0.552, 1.327, 1.465, 1.587, 0.407, P>0.05). The percentages of CD3⁺ T lymphocytes of children in group rhGH on the 3rd and 7th day of rhGH treatment were significantly higher than the percentage before rhGH treatment (P<0.05). (6) The mass concentration of serum IgA, complement C3, and IgG of children in the two groups was similar before and on the 3rd and 7th day of rhGH treatment (t=-1.596, -0.100, 1.263, -0.220, 1.378, 1.631, Z=0.228, 0.519, 1.182, P>0.05). The mass concentration of serum IgA and complement C3 of children in group rhGH on the 3rd and 7th day of rhGH treatment was significantly higher than that before rhGH treatment(P<0.05). Conclusions rhGH has little effect on humoral immunity of younger children with severe burn injuries with limited influence on CD19⁺ B lymphocyte, mass concentration of serum IgA, IgG, and complement C3. It may improve the cellular immunity function mainly through promoting the release of CD4⁺ T lymphocyte, reducing the release of CD8⁺ T lymphocyte. It can be used in clinical treatment of younger children with severe burn injuries.

Objective: To investigate the expression of gamma-aminobutyric acid type A receptor beta3 subunit (GABRB3) on cleft palate in C57BL/6J mice induced by 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD). Methods: Sixty C57BL/6J pregnant mice on gestation day (GD) 10.5 were divided into two groups: one group was administered through gastric tubes one dose of 28 μg/kg TCDD (experimental group) and the other group was administered through gastric tubes one dose of 5.6 ml/kg corn oil (control group). Embryos were removed by cesarean section from pregnant mice during the palatal formation stage (GD 13.5-17.5) and the palatal tissue studied in morphological and histological observation. The relative mRNA and protein expression of GABRB3 was measured by real-time quantitative PCR and Western blotting. Localization of GABRB3 protein was measured by immunohistochemistry or immunofluorescence. Results: The incidence of cleft palate at GD17.5 was 100% in experimental group and there was no cleft palate occurred in the control group (0); elevation of palatine processes in experimental group was completed on GD15.5 which was clearly delayed by a day compared with that in control group. On GD14.5-GD17.5, the mRNA expression (0.561±0.073, 0.728±0.104, 0.782±0.137, 0.686±0.145) and protein expression (0.288±0.013, 0.404±0.017, 0.399±0.012, 0.307±0.010) in the experimental group were significantly lower than the control group mRNA expression (0.818±0.088, 0.865±0.086, 1.021±0.054, 1.163±0.179) and protein expression (0.481±0.017, 0.456±0.009, 0.474±0.016, 0.529±0.015)(P<0.05). Immunohistochemistry and immunofluorescence showed that GABRB3 was mainly expressed in the mesenchymal cells and medial edge epithelium. Conclusions TCDD delayed palatal shelf elevation and eventually led to cleft palate may be associated with a decrease in GABRB3. GABRB3 may play an important role in the elevation and fusion phases of the palate development.

Objective:To explore the role of epithelial-mesenchymal transition(EMT) in fusion of the secondary palatal shelves to form the intact secondary palate induced by 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD).Methods:Twelve C57BL/6 J pregnant mice on gestation day (GD) 10.5 were divided into two groups: one group was conducted through gastric tubes with one dose of 28 μg/kg TCDD (experimental group) and the other group was operated through gastric tubes with equal volume corn oil (control group). Embryos were removed by cesarean section from pregnant mice during the palatal formation stage (GD 15.5) and the morphology of palatal tissue was observed. Primary media edge epithelial(MEE) were divided into experimental group and control group. MEE were treated with medium containing TCDD, 5 nmol/L, 10 nmol/L, 20 nmol/L and normal medium respectively. The expression of cytokeratin 19(CK-19) protein and vimentin protein in MEE were detected by immunofluorescence laser confocal microscopy and Western blotting after 72 hours. Statistical comparisons were made using one-way ANOVA.Results:A total of 36 fetuses were obtained in the experimental group, including 3 dead fetuses and absorbed fetuses. The incidence of cleft palate was 100% (33/33); the incidence of complete cleft palate was 84.8% (28/33), and the incidence of partial cleft palate was 15.2% (5/33); 40 fetuses were obtained in the control group, including 2 dead fetuses and resorbed fetuses, and the incidence of cleft palate was 0 (0/38). After 72 hours, the shape of MEE changed from uniform pebble-like to star-like or irregular shape with pseudopodia. The expressions of CK-19 protein were(0.739 ± 0.120, 0.483 ± 0.023, 1.007 ± 0.109, 1.086 ± 0.145) and fluorescence intensities were (53.384±5.785, 36.818 ± 8.250, 64.575±8.323, 76.898 ± 3.711) in control group and TCDD (5 nmol/L), TCDD (10 nmol/L) and TCDD (20 nmol/L) groups, respectively. The expressions of vimentin protein were (0.527 ± 0.112, 0.781 ± 0.095, 0.284 ± 0.046, 0.216 ± 0.040) and fluorescence intensities were (63.672±6.135, 82.632 ± 4.474, 52.608±7.525, 42.664 ± 7.659). Compared with the control group, the low-dose experimental group (5 nmol/L) had a decrease in CK-19 and an increase in vimentin; the high-dose experimental group (10 nmol/L, 20 nmol/L) had an increase in CK-19 and a decrease in vimentin, and the expression difference was statistically significant ( P<0.05), while there was no statistical significance among high-dose groups ( P>0.05). Conclusions:EMT process of MEE was identified in vitro and was a spontaneous procedure. TCDD-induced cleft palate may be related to the inhibition of the EMT process in MEE and with the increased dose of TCDD, the effects of EMT inhibiton were sustainable.

Objective:To explore the role of epithelial-mesenchymal transition(EMT) in fusion of the secondary palatal shelves to form the intact secondary palate induced by 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD).Methods:Twelve C57BL/6 J pregnant mice on gestation day (GD) 10.5 were divided into two groups: one group was conducted through gastric tubes with one dose of 28 μg/kg TCDD (experimental group) and the other group was operated through gastric tubes with equal volume corn oil (control group). Embryos were removed by cesarean section from pregnant mice during the palatal formation stage (GD 15.5) and the morphology of palatal tissue was observed. Primary media edge epithelial(MEE) were divided into experimental group and control group. MEE were treated with medium containing TCDD, 5 nmol/L, 10 nmol/L, 20 nmol/L and normal medium respectively. The expression of cytokeratin 19(CK-19) protein and vimentin protein in MEE were detected by immunofluorescence laser confocal microscopy and Western blotting after 72 hours. Statistical comparisons were made using one-way ANOVA.Results:A total of 36 fetuses were obtained in the experimental group, including 3 dead fetuses and absorbed fetuses. The incidence of cleft palate was 100% (33/33); the incidence of complete cleft palate was 84.8% (28/33), and the incidence of partial cleft palate was 15.2% (5/33); 40 fetuses were obtained in the control group, including 2 dead fetuses and resorbed fetuses, and the incidence of cleft palate was 0 (0/38). After 72 hours, the shape of MEE changed from uniform pebble-like to star-like or irregular shape with pseudopodia. The expressions of CK-19 protein were(0.739 ± 0.120, 0.483 ± 0.023, 1.007 ± 0.109, 1.086 ± 0.145) and fluorescence intensities were (53.384±5.785, 36.818 ± 8.250, 64.575±8.323, 76.898 ± 3.711) in control group and TCDD (5 nmol/L), TCDD (10 nmol/L) and TCDD (20 nmol/L) groups, respectively. The expressions of vimentin protein were (0.527 ± 0.112, 0.781 ± 0.095, 0.284 ± 0.046, 0.216 ± 0.040) and fluorescence intensities were (63.672±6.135, 82.632 ± 4.474, 52.608±7.525, 42.664 ± 7.659). Compared with the control group, the low-dose experimental group (5 nmol/L) had a decrease in CK-19 and an increase in vimentin; the high-dose experimental group (10 nmol/L, 20 nmol/L) had an increase in CK-19 and a decrease in vimentin, and the expression difference was statistically significant ( P<0.05), while there was no statistical significance among high-dose groups ( P>0.05). Conclusions:EMT process of MEE was identified in vitro and was a spontaneous procedure. TCDD-induced cleft palate may be related to the inhibition of the EMT process in MEE and with the increased dose of TCDD, the effects of EMT inhibiton were sustainable.