Objective:To observe the difference of the human telomeres RNA component (hTERC) genes’amplification in the cervical tissue by applying the environment-friendly fixative poly hydroxy acrylic acid and the transparent dewaxing solution Van-clear separately or jointly to replace the traditional fixative 4% (volume fraction)neutral buffered formalin and the conventional transparent dewaxing solu-tion xylene in the use of fluorescence in situ hybridization (FISH)for detection.Methods:In the study, 255 cases of cervical tissue specimens submitted by the Department of Gynecology in Zhongshan Boai Ho-sipital were collected from Mar.2013 to Apr.2015.Four samples were taken from the same lesion site. All the cases were divided into 4 groups and named group A,B,C,and D.Group A used 4% neutral buffered formalin fixed and xylene dewaxing to make slices.Group B used poly hydroxy acrylic fixed and xylene dewaxing to make slices.Group C used 4% neutral buffered formalin fixed and Van-clear trans-parent to make slices.Group D used poly hydroxy acrylic fixed and Van-clear transparent dewaxing to make slices.The amplification of hTERC genes in the four groups of cervical specimens was also detected by FISH technique.Results:When the hTERC genes were detected by FISH method under the fluore-scence microscope,it was obvious that the tissue profile and the background of group A,B,C and D were all clear.The probe was fixed in the accurate position so that the bright red or green fluorescence signals were easily found in these four groups.Compared with the positive rate of group A,there was no statistical significance in that of group B,C and D (P>0.05).At the same time,the coincidence rate of the FISH results was high,which showed that the new environment-friendly reagent had no significant difference in the detection of cervical hTERC genes by FISH technique.Conclusion:It is possible for the environment-friendly reagent poly hydroxy acrylic acid and Van-clear to replace 4%neutral buffered for-malin and xylene separately or jointly to detect the cervical hTERC genes by adopting FISH technique.

Background and purpose: Adequate tissue ifxation, transparent dewaxing is an important step of hematoxylin eosin (HE) staining and lfuorescence in situ hybridization (FISH) in detection of breast cancerHER-2 gene. The purpose of this study was to make a comparison between poly hydroxyl acrylic acid which is an environmen-tally friendly ifxation liquid and 4% neutral buffered formaldehyde in tissue ifxation for HE staining and FISH to detect theHER-2 gene in the breast cancer tissue sections. The study aimed to evaluate the feasibility of replacing 4% buffered formaldehyde, a traditional ifxation liquid, with the poly hydroxyl acrylic acid, an environmentally friendly ifxation lfuid.Methods:This project was performed on tissue samples collected from 69 cases of breast cancer, 41 cases of breast ifbroadenoma, 40 cases of uterine leiomyoma, 25 cases of cervical tissue, 25 cases of placenta obtained from the outpatient and inpatient departments of Zhongshan Boai Hospital from Mar. 2011 to Jan. 2015, from each of which two samples were drawn and two blocks of each specimen were divided into two groups randomly. Then one group was ifxed with 4% neutral buffered formaldehyde and made into 200 sections by HE while the other group was ifxed with poly hydroxyl acrylic acid and made into another 200 sections. The slice level of the two groups was determined by the staining condition of the sections, and SPSS 19.0 was employed to compare the excellent and good rate of HE staining. Additional 69 sections were produced with two groups of breast invasive ductal carcinoma tissues, and SPSS 19.0 was used to detect the ampliifcation ofHER-2 gene by FISH.Results:First, the number of best-quality slices stained with HE ifxed separately by poly hydroxyl acrylic acid and 4% neutral buffered formaldehyde was 155 and 166, respectively. The number of excellent pieces was 41 and 33, respectively, while the number of mediocre pieces was 3 and 1 with bad pieces being 1 and 0, respectively. The excellent and good rates of HE staining were 98% and 99.5%, respectively. There was no significant difference between the two groups (χ2=1.33,P>0.05).Second, the positive rates of the tis-sue slices by FISH ifxed separately by poly hydroxyl acrylic acid and 4% neutral buffered were 26.09% and 23.19%, respectively. There was no signiifcant difference between the two groups (χ2=0.50,P>0.05).Conclusion:The results obtained with HE staining and FISH using poly hydroxyl acrylic acid as a ifxation liquid are not signiifcantly different from those using 10% neutral buffered formaldehyde. Therefore, poly hydroxyl acrylic acid meets the requirements of environmental protection, and thus has the potential to be promoted and widely used.

Objective To compare two different methods to detect the differences of gene mutation rate, sensitivity, specificity and coincidence rate of epidermal growth factor receptor (EGFR) in non-small-cell lung cancer (NSCLC) so as to assess the clinical value of qRT-PCR method and its environmental-friendly technologyplatforms.One uses environmental fixative poly hydroxyl acrylic acid and green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ (volume fraction) neutral buffered formalin and the traditional transparent dewaxing liquid xylene in application of quantitative real-time polymerase chain reaction (qRT-PCR).The other uses traditional reagents in direct sequencing.Methods We selected 91 cases of primary NSCLC specimens resected between May 2013 and March 2016 in Zhongshan Bo`ai Hospital and Zhongshan Hospital of Traditional Chinese Medicine.Five samples were taken from the same tumor lesion.We used a random number table to randomly divide these samples into Groups A, B , C, D, and E.Group A received direct sequencing method in detection of EGFR gene mutations.Besides, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group B received qRT-PCR method to detect EGFR gene mutations.Meanwhile, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group C received qRT-PCR method in detection of EGFR gene mutations.At the same time, during the experiment, polyhydroxy acrylic acid was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group D received qRT-PCR method to detect EGFR gene mutations.In the meantime, 4％ neutral buffered formalin was used for fixing, Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.Group E received qRT-PCR method in detection of EGFR gene mutations.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.The mutations of Exons 18, 19, 20, and 21 in EGFR genes were respectively determined in the five groups of NSCLC.Results ① Groups B, C, D, E and A did not significantly differ in the percentage of people with mutations or target site mutation rates of EGFR genes in NSCLC (P> 0.05).② The detection results of EGFR target site mutation in Groups B, C, D, E and A had good sensitivity, strong specificity, and high compliance rate.Conclusion The green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ neutral buffered formalin and the traditional transparent dewaxing liquid xylene in the application of qRT-PCR so as to detect EGFR gene mutations in NSCLC has good consistent results compared with the method that uses traditional reagents in direct sequencing.It has the significance and value in clinical application.

Objective: To analyze the characteristics and treatment of diabetic patients with superficial partial-thickness burn on feet. Methods: Eighty-three patients with superficial partial-thickness burn on 119 feet were hospitalized in our unit from January 2011 to December 2017. The medical records of the patients with 46 men and 37 women, aged 60±11 were retrospectively analyzed. The patients were divided into diabetes group and non-diabetes group according to whether they had diabetes or not, with 41 patients (60 burn feet) in diabetes group and 42 patients (59 burn feet) in non-diabetes group. Patients in diabetes group and non-diabetes group were given systemic treatment and wound dressing change. Thirty-seven diabetic patients whose wounds deepened to deep partial-thickness were divided into eschar shaving group and non-eschar shaving group according to patients′ willingness and the treatment, with 14 patients in eschar shaving group and 23 patients in non-eschar shaving group. Patients in eschar shaving group were given eschar shaving operation at early stage, and patients in non-eschar shaving group were given wound dressing change. The length of hospital stay, hospitalization treatment expenses, pulse of arteria dorsal pedis and posterior tibial artery immediately after admission, deepening of wounds on feet during hospital stay, and rates of wound healing on feet of patients in diabetes group and non-diabetes group were observed and calculated. Pulses of arteria dorsal pedis and posterior tibial artery immediately after admission, deepening of wounds on feet during hospital stay, positive rates of bacteria and fungus in wounds on feet, and rates of wound healing on feet of patients in eschar shaving group and non-eschar shaving group were observed and calculated. Data were processed with chi-square test, t test, Fisher′s exact propability method, and Mann-Whitney U test. Results: The length of hospital stay of patients in diabetes group was (29±20) d, which was significantly longer than that of patients in non-diabetes group [(19±13) d, t=2.730, P<0.01]. The hospitalization treatment expense of patients in diabetes group was (46 988±41 322) yuan, which was significantly more than that of patients in non-diabetes group [(29 106±24 813) yuan, t=2.396, P<0.05]. The pulses of arteria dorsal pedis and posterior tibial artery of patients in diabetes group were significantly weaker than those of patients in non-diabetes group (Z=3.278, 2.194, P<0.05 or P<0.01). The percentages of wounds on feet of patients in diabetes group deepening to deep partial-thickness burn, full-thickness skin defect with bone and tendon exposure were respectively 88.3% (53/60) and 23.3% (14/60), which were significantly higher than those of patients in non-diabetes group [47.5% (28/59) and 1.7% (1/59), χ²=22.867, 12.644, P<0.01]. Rate of wound healing on feet of patients in diabetes group was 78.3% (47/60), which was significantly lower than 100.0% (59/59) of patients in non-diabetes group ( χ²=14.351, P<0.01). There were respectively 21 and 32 feet in patients of eschar shaving group and non-eschar shaving group. There were no significantly statistical differences in pulses of arteria dorsal pedis and posterior tibial artery of patients between eschar shaving group and non-eschar shaving group (Z=0, 0.453, P>0.05). The percentage of wounds on feet of patients in non-eschar shaving group deepening to full-thickness skin defect with bone and tendon exposure was 43.8% (14/32), which was significantly higher than 0 of patients in eschar shaving group ( χ²=12.486, P<0.01). Positive rates of bacteria and fungus in wounds on feet of patients in eschar shaving group was significantly lower than that of patients in non-eschar shaving group (χ²=4.386, P<0.05 ). Rate of wound healing on feet of patients in non-eschar shaving group was 59.4% (19/32), which was significantly lower than that of patients in eschar shaving group [100.0% (21/21), P<0.01]. Conclusions Diabetes patients with superficial partial-thickness burn wounds on feet has long length of hospital stay, high hospitalization treatment expenses. Wounds of the patients are easy to deepen, with low wound healing rate. Eschar shaving at early stage when the wounds deepened to deep partial-thickness burn is a good way to increase wound healing rate and prevent further deepening of wounds.

OBJECTIVE: To investigate paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples. METHODS: p16 immunohistochemical staining was performed in 150 cases of chronic cervicitis, 126 cases of low-grade squamous intraepithelial lesions(LSIL), 96 cases of high-grade squamous intraepithelial lesions (HSIL) and 78 cases of cervical cancer from January 2014 to March 2018 in Zhongshan Boai Hospital. The results of p16 protein expression in paraffin sections with thickness of 2, 3, 4, 5 and 6 μm were compared using Logistic regression analysis. RESULTS: With the increase of slice thickness, the staining of p16 protein in nucleus gradually increased. The thickness of cervical slices in chronic cervicitis and cervical cancer samples had no significant effect on the positive rate of p16 protein(=7.817 and 1.332, both >0.05), while the thickness of slices in LSIL and HSIL samples had significant effect on the positive rate of p16 protein (=17.688 and 10.182, <0.05 or <0.01). The stable and reliable results were obtained when the slices were between 3 and 5 μm thick. CONCLUSIONS Paraffin sections with thickness of 3.0-5.0 μm are recommended for immnohistochemical staining of p16 protein in cervical tissue samples.
Biomarkers, Tumor ; genetics ; metabolism ; Cervical Intraepithelial Neoplasia ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Female ; Histocytological Preparation Techniques ; standards ; Humans ; Immunohistochemistry ; Paraffin ; Squamous Intraepithelial Lesions of the Cervix ; Uterine Cervical Neoplasms ; physiopathology

OBJECTIVETo study the high risk factors of allergen sensitization among 1-16 years old children with allergic rhinitis (AR) symptoms.

METHODSMedical history was collected from 518 children with AR symptoms aged 1 to 16 years old between April 2011 and November 2012, including five aspects:basic information, disease characteristics, associated with other allergic diseases, growth and development history and genetic history and so on. The allergens were determined by UniCAP 100 system. The high risk factors of allergen sensitization (sIgE ≥ grade III) among 1-16 years old children with AR symptoms were analyzed. The data processing and statistical analysis were conducted by SPSS 17.0 software.

RESULTSThree hundred and thirty of 518 patients (63.7%) could be diagnosed as AR. The primary allergen was mixed fungal (52.4%). The highest percentage of allergen sIgE ≥ grade III accounted for the corresponding allergen positive cases was 71.1% (mixed fungal). Single factor analysis of clinical characteristics in the groups of AR and non AR showed that the history of months(χ(2) = -3.591), concomitant allergic conjunctivitis (χ(2) = 12.325) and cats or dogs at home (χ(2) = 5.615) were statistically significant between the two groups (all P < 0.05).In children with AR, single factor analysis of clinical characteristics in the groups of whether positive allergen level ≥ grade III showed that the concomitant asthma(χ(2) = 4.097), breastfeeding(χ(2) = 4.186), the housing situation (χ(2) = 4.360) and the bedroom toward (χ(2) = 4.656) were statistically significant between the two groups (all P < 0.05). In children with AR, single factor analysis of clinical characteristics in different age groups showed that the history of months(χ(2) = 64.999), concomitant eczema (χ(2) = 24.056), concomitant insect bite dermatitis (χ(2) = 9.148), cats or dogs at home (χ(2) = 8.529) and mother suffered from AR (χ(2) = 9.565) were statistically significant in different age groups (all P < 0.05).

CONCLUSIONThe study revealed that the longer of history, concomitant allergic conjunctivitis and cats or dogs at home are risk factors for AR;In children with AR, not breastfeeding and the bedroom toward back are risk factors for inhalation allergen sIgE ≥ gradeIII.

Adolescent ; Allergens ; immunology ; Animals ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulin E ; immunology ; Infant ; Male ; Pets ; Rhinitis, Allergic ; etiology ; immunology ; Risk Factors ; Skin Tests