BACKGROUND:There are few studies concerning estrogen receptorβgene, and its mechanism of regulating the bone metabolism is stil unclear now. OBJECTIVE:To analyze the effect of estrogen receptorβ(ERβ) silencing on the expressions of transforming growth factorβ1 (TGF-β1) and bone morphogenetic protein 2 (BMP-2) in human osteoblasts METHODS:There were three groups:blank control group (hFOB 1.19 uninfected with any retrovirus);negative control group (containing invalid interference fragment ERβ-shRNA-nc);optimal RNAi group (ERβ-shRNA-3). ERβ-shRNA retroviral vectors in the optimal RNAi group were used to transfect human osteoblasts fol owed by resistance screening and cel expansion. MTT assay was used to detect the proliferative activity of ERβ-silenced osteoblasts. Then under estrogen intervention, the stable inhibition rate of ERβwas determined using western blot assay, and the expressions of TGF-β1 and BMP-2 in human osteoblasts after ERβsilencing were detected by RT-PCR technology and western blot assay. RESULTS AND CONCLUSION:Human osteoblasts that were stably transfected by ERβ-shRNA-3 retroviral vector was selected successful y, and ERβsilencing had no significant influence on the cel proliferation (P>0.05). Under the interference of estrogen, the silencing efficiency of ERβprotein was (93.11±0.57)%(P<0.05), and after ERβsilencing, the expressions of TGF-β1 and BMP-2 were increased by (26.65±3.81)%and (16.62±1.71)%at mRNA level, and increased by (23.79±3.76)%and (18.08±3.20)%at protein level (both P<0.05). In conclusion, ERβmay play an important role in bone metabolism by regulating the expressions of TGF-β1 and BMP-2.

Objective To explore the working pattern for endocrinology clinical pharmacists participating in clinical drug treatment .Methods By participating in the treatment for a patient with diabetes and hyperprolactinemia and analyzing the cau‐ses of poor glycemic control and hyperprolactinemia ,clinical pharmacists proposed therapeutic regimen .Results The long‐term psychotropic medication may be one of the reasons why blood glucose of the patient was unsatisfactorily controlled .The patient with hyperprolactinemia may be associated with her longstanding use of paliperidone .The patient was recommended to consult a psychiatrist and switch to an alternative medication which does not cause hyperprolactinemia .Conclusion The active partici‐pation of clinical pharmacists during clinical drug therapy could improve the medication rationality and compliance of patients with drugs .

BACKGROUND:Studies concerning how estrogen receptorβparticipates in bone metabolism are few now. OBJECTIVE:To investigate the effect of estrogen receptorβon the expression of osteoprotegerin and receptor activator of nuclear factor-κB ligand in human osteblast-like cells. METHODS:The retrovirus with the most effective interference sequence and non-specific short hairpin RNA was used to transfect human osteoblast-like cellMG63 in order to screen out the stable colon, and then amplified and cultured. The blank control and non-specific short hairpin RNA were used as control, and the stable inhibition rate of estrogen receptorβwas detected. The 17β-estradiol was added into the cells in three groups, that were MG63 cells, short hairpin RNA retrovirus estrogen receptorβ-mediated MG63 cells and negative control short hairpin RNA retrovirus-medicated MG63 cells, in order to detect the expressions of osteoprotegerin and receptor activator of nuclear factor-κB ligand mRNA in human osteoblast-like cells. RESULTS AND CONCLUSION: The human osteoblast-like MG63 cellline was further stably transfected with pRNAT-H1.4/Retro-estrogen receptorβshort hairpin RNA3, and then compared with the blank control and negative control, and found that estrogen receptorβcould express the stable inhibited human osteoblast-like cellline. The inhibition rate of estrogen receptorβmRNA was (88.17±1.17)%(P<0.05), and the inhibition rate of estrogen receptorβprotein was (89.01±1.22)%(P<0.05), indicating that estrogen receptorβgene knockdown human osteoblast-like cellmodels were constructed successful y. After estrogen intervention for 48 hours, the inhibition of MG63 cells with estrogen receptorβcould up-regulate the osteoprotegerin mRNA and protein expression in the blank control group and the negative control group (P<0.05), down-regulate the receptor activator of nuclear factor-κB ligand mRNA and protein expression (P<0.05), and up-regulate the osteoprotegerin receptor activator of nuclear factor-κB ligand expression. The results indicate that estrogen receptorβmay play an important role in bone metabolism through regulating osteoprotegerin/receptor activator of nuclear factor-κB ligand ratio.

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 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.

ObjectiveTo find out the risk factors causing iatrogenic spinal cord injury (ISCI) so as to provide theoretical support for reducing the spinal cord injury during spinal operation. Methods A retrospective study was done on 120 patients undergone cervical or thoracic spinal( C1-T12 ) surgery at Xiangya Hospital of Central South University from January 2002 to January 2009. The patients were randomly divided into injury group (n = 34) and control group (n = 86) and the univariate analysis was used to analyze 30 factors including clinical factors, iconography factors, operation and pathology factors as well as possible protective factors. Then, the factors with statistical difference were analyzed by using the multi-factor unconditioned Logistic analysis.Results The univariate comparison between the two groups showed statistical difference ( P ＜ 0. 05 ) in nine factors including combined hypertension, combined diabetes mellitus, preoperative ASIA grade, spinal canal stenosis rate, ratio of spinal cord area/efficient area of vertebral canal, spinal cord MRI T2WI high signal, bleeding amount during operation, intraspinal prominence adhesion to dura mate of spinal cord as well as intraoperative use of methylprednisolone. The multi-factor Logistic regression analysis revealed that ASIA grade, value of spinal cord area/efficient area of vertebral canal, spinal cord MRI T2W1 high signal and bleeding amount in operation had positive correlation with ISCI. Use of methylprednisolone during operation had negative correlation with ISCI. ConclusionsCombined diabetes mellitus, ASIA grade, spinal cord MRI T2W1 high signal, ratio of spinal cord/vertebral canal area and bleeding amount in operation are the risk factors for ISCI. Use of large dose methylprednisolone exerts preventive effect on ISCI.

BACKGROUND:The monocyte chemoattactant protein-1 gene polymorphism is associated with spinal tuberculosis susceptibility. OBJECTIVE:To investigate the association between serum monocyte chemoattactant protein-1 expression level and spinal tuberculosis susceptibility in Han population of Hunan province. METHODS:The patients with spinal tuberculosis and the healthy volunteers were recruited in Xiangya Hospital of Central South University from December 2004 to December 2010. The empty peripheral venous blood 2 mL were col ected from the subjects in early morning, then the monocyte chemoattactant protein-1-362 genotypes were detected by polymerase chain reaction and DNA sequencing technology. And the serum monocyte chemoattactant protein-1 level was detected by enzyme linked immunosorbent assay technology. The ROC curve was used for diagnostic tests to calculate diagnostic threshold value of serum monocyte chemoattactant protein-1 level to spinal tuberculosis susceptibility, and to analyze the diagnostic titer. RESULTS AND CONCLUSION:208 patients with spinal tuberculosis and 210 healthy volunteers were included. The serum monocyte chemoattactant protein-1 level of the spinal tuberculosis patients was significantly higher than that of the healthy volunteers [(134.58±51.63) ng/L vs. (39.18±17.45) ng/L, P<0.01]. The serum monocyte chemoattactant protein-1 level could not be affected by gender, but over-expressed in patients with monocyte chemoattactant protein-1-362-CC genotypes. The serum monocyte chemoattactant protein-1 level higher than 101.65 ng/L indicated that the patients might suffered from spinal tuberculosis (sensitivity:85.5%, specificity:94.3%, Youden index:0.799, area under curve of ROC:0.946, 95%confidence interval:0.916-0.975, P<0.01). The serum monocyte chemoattactant protein-1 level may be associated with spinal tuberculosis susceptibility in Han population of Hunan province, highly expressed serum monocyte chemoattactant protein-1 can be used as one of the indicators for the diagnosis of spinal tuberculosis.

Objective To explore the correlative factors that affected the early clinical efficacy of surgical management of lumbar disc herniation.Method 208 cases of lumbar disc herniation were recruited since December 2007.The details of their therapy in different periods were compared and analyzed.Result The aggressive discectomy was the most powerful factor related to the better early clinical outcome.The patients with preoperative JOA score ＞ 17 were associated to the poor clinical outcome.The patients with postoperative JOA score ≥ 25 on 3 month and ≥ 24 on 1 year after operations were associated to better early clinlcal outcome.Conclusion The pre- and post-operative JOA score and aggressive discectomy were the factors affected the clinical outcome.

Objective:To investigate the accuracy and feasibility of 3D-printing individualized template-guided and CT-guided 192Ir interstitial brachytherapy in the central recurrent gynecologic tumors by comparing pre-plan and intraoperative physical dosimetric parameters. Methods:This study involved 38 patients with central recurrent gynecologic tumors who underwent 3D printing individual template (3D-PIT)-assisted and CT-guided 192Ir interstitial brachytherapy in the Department of Radiation Oncology of the Peking University Third Hospital from Jan 2018 to Dec 2019.The prescription doses for the target tumor areas were 10-36 Gy to be delivered at 5-6 Gy/fraction for 2-6 fractions.The pre-plan and intraoperative dosimetric parameters were compared, including the minimum prescription doses delivered to 90% and 100% of target volume( D90, D100)and the mean percentage of volume receiving 100% of the prescription doses ( V100). Meanwhile, the doses delivered to 2 cm 3 ( D2 cm 3) of organs at risk (bladders, rectums, and colons) were analyzed.The quality parameters of the brachytherapy were studied, including conformity index (CI), homogeneity index (HI), and external index (EI) of the target volume.Perioperative complications were also observed. Results:A total of 194 treatments were included.During the treatment, 5-13 (median 6) needles were inserted, with a prescription dose of 5-6 Gy per fraction.There were no statistical differences between pre-plan and intraoperative D90, D100, V100, CI, HI, and EI as well as the D2 cm 3 of bladders and colons at risk ( P>0.05). In contrast, for the D2 cm 3 of rectums, the intraoperative dose was slightly higher than the pre-plan dose, showing a statistical difference ( t=-0.335, P=0.027). Conclusions:The 3D-PIT-assisted and CT-guided 192Ir interstitial brachytherapy at a high dose rate is accurate and feasible in the treatment of recurrent gynecologic tumors, meeting the pre-plan dose requirement.

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.