Objective:To investigate the effect of miR-27b-3p on radiation resistance of breast cancer cells.Methods:The relative expression levels of miR-27b-3p in normal tissues and breast cancer tissues were analyzed through GEO database and verified by the qRT-PCR assay. Cloning formation, immunofluorescence, and EDU assay were used to assess the functions of miR-27b-3p on radioresistance of breast cancer cells. The luciferase reporter assay was used to verify whether miR-27b-3p directly targeted PLK2 mRNA. A rescue experiment was performed to identify the influence of PLK2 overexpression on miR-27b-3p regulated radioresistance.Results:The expression level of miR-27b-3p in both breast cancer tissues ( t=2.99, P<0.01) and breast cancer cells ( t=21.21, 32.88, P<0.05) was significantly higher than those in normal breast tissues and cells, especially, it was elevated in radioresistant MCF-7R cells ( t=25.63, P<0.05). Overexpression of miR-27b-3p enhanced the cloning efficiency of MCF-7 cells ( t=10.32, P<0.05), and had a protective effect on the proliferation of irradiated MCF-7 cells ( t=8.77, 8.26, 8.03, P<0.05). But interference of miR-27b-3p reduced the cloning efficiency and proliferation of MCF-7R cells ( t=40.00, P<0.05) after irradiation with different doses( t=8.54, 8.32, 8.23, P<0.05). Moreover, PLK2 was verified to be a direct target of miR-27b-3p, and overexpression of PLK2 inhibited miR-27b-3p-mediated radioresistance of breast cancer cells (MCF-7: t=9.66, P<0.05; MCF-7R: t=6.42, P<0.05). Conclusions:miR-27b-3p contributes to the radioresistance of breast cancer cells by targeting PLK2.

Objective:To explore the efficacy and safety of anti-B cell maturation antigen (BCMA) chimeric antigen receptor T-cell (CAR-T) for the retreatment of relapsed and refractory multiple myeloma (RRMM).Methods:The clinical data of 10 RRMM patients who received anti-BCMA CAR-T therapy for the second time (CART2) in Henan Province Hospital of Traditional Chinese Medicine due to failure or recurrence after their first anti-BCMA CAR-T (CART1) therapy from January 2017 to June 2021 were retrospectively analyzed. The treatment, efficacy and adverse events of patients receiving CART2 therapy were summarized; and the objective response rate (ORR), median duration of response (DOR) and incidence of adverse reactions were compared between CART1 and CART2.Results:Among 10 patients, 8 were males and 2 were females, with a median age of 57 years (41-70 years). Patients' 3-month ORR after CART1 therapy was 90%, and the median DOR was 16.0 months (3.0-27.0 months). CART2 used human-derived anti-BCMA CAR-T to treat 6 cases and mouse-derived anti-BCMA CAR-T to treat 4 cases. The 3-month ORR of patients receiving CART2 therapy was 40%, and the median DOR was 8.5 months (3.0-11.0 months). Among 9 patients who received mouse-derived anti-BCMA CAR-T in CART1 therapy, 4 of them received the same product again and none of them showed curative effect. Among 6 patients retreated with human-derived anti-BCMA CAR-T, 4 patients (66.7%) of them achieved partial remission (PR) or better. During CART1 therapy, 10 patients developed grade 1-2 cytokine release syndrome (CRS), and 7 patients developed different degrees of decrease in leukocyte, neutrophil absolute count (ANC) and platelet. Among patients who achieved effective outcomes after receiving CART2 therapy, 4 patients of them developed grade 1-2 CRS, and different degrees of decrease in white blood cell, ANC and thrombocytopenia. Immune effector cell-related neurotoxicity syndrome was not observed.Conclusions:Anti-BCMA CAR-T is effective and safe to retreat RRMM. The ORR and DOR of patients receiving CART2 therapy are lower than those of patients receiving CART1 therapy. CRS and cytopenia are common adverse reactions.

Objective:To explore the clinical effect of lenalidomide combined with anti-B-cell maturation antigen chimeric antigen receptor T cell (anti-BCMA CAR-T) in the treatment of relapsed/refractory multiple myeloma (RRMM).Methods:The clinical data of a patient with RRMM who underwent lenalidomide combined with anti-BCMA CAR-T therapy in Henan Province Hospital of Traditional Chinese Medicine in January 2020 were analyzed. Clinical manifestations, diagnosis and treatment were also analyzed, and the related literature was reviewed.Results:The patient was a 51-year-old man who was diagnosed as IgD-λ multiple myeloma (MM) in October 2015. The patient achieved remission after 10 courses of chemotherapy regimens including immunomodulators and proteasome inhibitors, followed by autologous hematopoietic stem cell transplantation. MM relapsed after 14 months of transplantation. His disease continued to progress after multiple chemotherapy regimens and mouse or human-derived anti-BCMA CAR-T therapy. After a conditioning chemotherapy regimen of fludarabine and cyclophosphamide, the patient took lenalidomide on day 1 and was infused human-derived anti-BCMA CAR-T cells on the next day. Grade 3 cytokine releasing syndrome (CRS) appeared after infusion, and was resolved after symptomatic treatment. Very good partial response (VGPR) was achieved on day 14 after anti-BCMA CAR-T treatment. VGPR had been maintained for more than 3 months by press time.Conclusion:Lenalidomide combined with anti-BCMA CAR-T therapy is feasible and effective in the treatment of RRMM.

Objective:To study the incidence of glucose-6-phosphate dehydrogenase (G6PD) deficiency and the gene carrying status of newborns in Guangxi Zhuang Autonomous Region (Guangxi for short), so as to provide theoretical basis for clinical genetic counseling and accurate diagnosis.Methods:A total of 63 606 newborns who underwent G6PD screening in Guangxi Neonatal Disease Screening Center from January 2017 to December 2018 were selected as study subjects; heel blood was collected to prepare dry blood spots. Fluorescence quantitative analysis was used in the preliminary screening, and the newborns with positive preliminary screening were recalled by telephone; further diagnosis was carried out via the G6PD/6-phosphogluconate dehydrogenase (6PGD) ratio method and genetic testing, the diagnosis rate of the two methods of newborns with positive preliminary screening were compared and analyzed, and genetic mutation testing was conducted.Results:Among 63 606 newborns who underwent G6PD preliminary screening, 4 267 newborns with G6PD positive were detected, and the positive rate of preliminary screening was 6.7%. Among them, the positive rates of preliminary screening of males and females were 10.3% (3 508/33 988) and 2.6% (759/29 618), respectively. The positive rate of preliminary screening of males was significantly higher than that of females ( P < 0.01). A comparative analysis of 777 newborns (519 males and 258 females) that underwent G6PD/6PGD ratio method and genetic testing at the same time as the recall showed that the diagnosis rate of the two methods for male newborns was the same, both of which were 95.6% (496/519). Among female newborns, 168 and 236 confirmed cases were detected by G6PD/6PGD ratio method and genetic testing, respectively, and the diagnosis rates were 65.1% (168/258) and 91.5% (236/258), respectively. The results of genetic mutation testing showed that the five common genotypes in Guangxi were c.1388 G>A, c.1376 G>T, c.95 A>G, c.871 G>A, and c.1024 C>T, respectively. Conclusions:The positive rate of G6PD preliminary screening of newborns in Guangxi is relatively high. It is recommended that G6PD/6PGD ratio method and genetic testing should be performed at the same time for diagnosis of female newborns with positive preliminary screening to avoid missed diagnosis and misdiagnosis.

Clinical final examination is an important link in the standardized training of residents in dermatology and quantification of evaluation indicators is one of the important parameters. In this study, the scores of clinical examinations of six candidates who participated in standardized training of residents in dermatology of Sun Yat-Sen University in June 2019 were taken as examples to explore the quantitative indicators of standardized and multi-station clinical final examinations. The indicators contained four stations and five links: skin pathological reading, skin biopsy, medical history collection and physical examination, medical record writing, and comprehensive questioning, which covered the main contents of the standardized training outline of residents in dermatology. Each evaluation indicator was refined and quantified. Finally, heuristic ideas were put forward, including a wider range of standardized and multi-station clinical examinations, introduction of new examination places, and utilization of information technology.

Tissue inhibitor of metalloproteinases-2 (TIMP-2) inhibits tumor migration and invasion. Obtaining TIMP-2 protein is conducive to a comprehensive and in-depth study of its function and mechanism in tumorigenesis and development. We collected human TIMP-2 protein through prokaryotic expression in vitro. We expressed, purified and characterized human TIMP-2 protein. First, the human TIMP-2 gene was cloned from the cDNA obtained by reverse transcription of total RNA of human lung cancer A549 cells, and constructed to pET28a vector. The recombinant plasmid pET28a-TIMP-2 was transformed into Escherichia coli BL21(DE3) after restriction endonuclease digestion and sequencing analysis. The expression of TIMP-2 protein was induced by isopropyl-β-D-thiogalactoside (IPTG), and the expression conditions were optimized. After purification by nickel affinity column, the fusion protein His-TIMP-2 was identified by Western blotting method and its biological activity was detected by gelatin zymography. The fusion protein His-TIMP-2 existed in the form of inclusion body in E. coli. In a certain range, the concentration of IPTG had no significant effect on the expression amount of His-TIMP-2. But in this expression system, induction temperature and time were the key parameters, and the expression amount of His-TIMP-2 in E. coli increased with the increase of induction temperature. The purified and refolded fusion protein could effectively inhibit the activity of matrix metalloproteinases expressed by human lung cancer A549 cells. The acquisition of active fusion protein lays a foundation for further study of the function and mechanism of human TIMP-2, and is of great significance for tumor therapy.
Cloning, Molecular ; Escherichia coli/genetics* ; Humans ; Recombinant Fusion Proteins/genetics* ; Recombinant Proteins ; Tissue Inhibitor of Metalloproteinase-2/genetics*

Taking the course's characteristics and advantages into account, this study explores the systematic implementation of standardized teaching rounds in the clinical probation sessions of the Dermatology and Venereology course , and evaluates its effect by analyzing the cases of 125 medical students from grade 2015 of Sun Yat-sen University . The results showed that the implementation of standardized teaching rounds has positive effect on the course.

Objective: To explore the clinical application value of enhanced recovery after surgery (ERAS) in the laparoscopic surgery for cholecystolithiasis comorbid with choledocholithiasis. Methods: The prospective study was conducted. The clinicopathological data of 52 patients with cholecystolithiasis comorbid with choledocholithiasis who were admitted to the Third Affiliated Hospital of Zunyi Medical University from September 2016 to September 2018 were collected. Patients were divided into 2 groups by random number table: patients in observation group received laparoscopic cholecystectomy + choledocholithotomy + choledochoscopic exploration + T-tube drainage (or primary suture of common bile duct) and perioperative management guided by the concept of enhanced recovery after surgery (ERAS), and patients in control group received traditional perioperative management. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) postoperative complications; (4) postoperative pain scores; (5) changes in hepatic function and blood routine during perioperative period. Follow-up using outpatient examination and telephone interview was performed to detect complications during the postoperative 6 months up to March 2019. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was analyzed using the paired t test or repeated ANOVA. Count data were described as absolute numbers and percentages, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Results: Fifty-two patients were screened for eligibility, including 20 males and 32 females, aged 25-68 years, with an average age of 53 years. There were 30 patients in the observation group and 22 in the control group. (1) Surgical situations: the operation time and volume of intraoperative blood loss were (133±19)minutes and (47±21)mL in the observation group, and (136±22)minutes and (49±23)mL in the control group, respectively, showing no significant difference between the two groups (t=-0.386, -0.211, P>0.05). (2) Postoperative situations: time to out-of-bed activity, time to initial food intake, time to first anal flatus, duration of postoperative hospital stay, and hospital expenses were (18±4)hours, (19±5)hours, (28±2)hours, (4.0±1.0)days, and (1.82±0.22)×10⁴ yuan in the observation group, and (29±7)hours, (46±9)hours, (37±4)hours, (6.6±1.6)days, and (2.25±0.29)×10⁴ yuan in the control group, respectively, showing significant differences between the two groups (t=-7.054, -14.169, -9.426, -6.582, -5.809, P<0.05). (3) Postoperative complications: 1 of 30 patients in the observation group had postoperative biliary leakage, with a postoperative complication rate of 3.3%, and was cured after symptomatic support treatment. Six of 22 patients in the control group had postoperative complication, with a postoperative complication rate of 27.3%, including 2 of biliary leakage, 1 of hemorrhage, 1 of abdominal infection, 1 of pulmonary infection, 1 of urinary infection, and they were cured after symptomatic support treatment. There was a significant difference between the two groups (χ²=4.358, P<0.05). (4) Postoperative pain scores: from postoperative 6 hours to 48 hours, the postoperative pain score changed from 2.4±0.7 to 1.9±0.9 in the observation group, and from 4.1±0.7 to 2.9±0.9 in the control group, respectively, showing a significant difference in the changing trend between the two groups (F=78.053, P<0.05). (5) Changes in hepatic function and blood routine during perioperative period: from preoperation to postoperative 3 days, levels of alamine aminotransferase (ALT), aspartate transaminase (AST), gamma-glutamyltransferase (GGT), total bilirubin (TBil), and count of white blood cells in the observation group changed from (77±20)U/L to (53±12)U/L, from (85±22)U/L to (61±17)U/L, from (166±39)U/L to (55±24)U/L, from (40±13)μmol/L to (29±12)μmol/L, from (7.0±2.0)×10⁹/L to (6.8±1.9)×10⁹/L, and changed from (79±23)U/L to (62±14)U/L, from (88±24)U/L to (64±17)U/L, from (179±34)U/L to (74±29)U/L, from (45±13)μmol/L to (35±12)μmol/L, from (7.9±2.4)×10⁹/L to (7.5±1.9)×10⁹/L in the control group, respectively. The levels of ALT, AST, GGT, TBiL, and count of WBC showed increasing at postoperative 1 day, and decreasing at postoperative 3 days. There was no significant difference in the changing trend between the two groups (F=0.058, 0.471, 3.021, 1.593, 2.172, P>0.05). Conclusion ERAS is safe and effective in the laparoscopic surgery for choledocholithiasis comorbid with cholecystolithiasis.

Objective: To analyze changes in sensitivity of clinical strains of Ureaplasma urealyticum (Uu) to 9 kinds of common antibiotics from 2017 to 2018. Methods: The results of drug sensitivity testing of clinical Uu strains, which were isolated from 19 431 patients in Laboratory of Sexually Transmitted Disease (STD) , Department of Dermatology and Venereology, The Third Affiliated Hospital, Sun Yat-Sen University from 2007 to 2018, were analyzed retrospectively. Of the 19 431 patients, 6 076 were males, and 13 355 were females. Their age ranged from 15 to 68 years. The tested antibiotics included minocycline, doxycycline, erythromycin, azithromycin, clarithromycin, roxithromycin, ciprofloxacin, sparfloxacin and levofloxacin. Results: Resistance rates of clinical Uu strains to minocycline and doxycycline gradually decreased from 10.08% and 10.42% in 2007 to 1.15% and 2.61% in 2018, respectively, while sensitivity rates to minocycline and doxycycline gradually increased from 85.88% and 87.56% in 2007 to 97.02% and 96.42% in 2018, respectively. The resistance rate and sensitivity rate of clinical Uu strains to erythromycin fluctuated greatly during 2014—2017, with the resistance rate fluctuating around 20%, and the sensitivity rate fluctuating around 50%. The resistance rate of clinical Uu strains to azithromycin dropped rapidly from 42.02% in 2007 to 8.39% in 2011, and then fluctuated slightly around 10%. However, the sensitivity rate to azithromycin increased from 8.40% in 2007 to 86.05% in2011, and remained above 80% from then on to 2018. During 2007—2018, Uu strains showed low resistance rates (10%-20%) and high sensitivity rates to clarithromycin (80%-90%) , and the resistance and sensitivity rates to roxithromycin were similar to those to erythromycin. Uu strains showed constantly high resistance to ciprofloxacin (more than 80% after 2013) and low sensitivity (persistently less than 10%) . The sensitivity rate of clinical Uu strains to sparfloxacin fluctuated around 40%, while the resistance rate was maintained below 10% after 2011. The resistance and sensitivity rates of Uu strains to levofloxacin were similar to sparfloxacin, but the sensitivity rate to levofloxacin was relatively lower, which had been maintained at about 30%. Conclusion From 2007 to 2018, clinical Uu strains maintained a relatively stable low resistance rate and a high sensitivity rate to minocycline, doxycycline and clarithromycin, a high resistance rate and low sensitivity rate to ciprofloxacin, and a low resistance rate and sensitivity rate to sparfloxacin and levofloxacin; a relatively stable low resistance rate and a high sensitivity rate to azithromycin were achieved only after 2011; the resistance rate and sensitivity rate to erythromycin and roxithromycin fluctuated greatly.

A total of 105 undergraduate students of the class 2014 majoring in clinical medicine in Sun Yat-Sen University received probation teaching in our department from October 2017 to May 2018,according to the syllabus of dermatology and venereology of clinical medicine.Senior teachers in our department were responsible for probation teaching,with the help of the independent wards in our department and probation teaching at the outpatient service.The teachers recorded the whole teaching process and evaluated the students' memory after probation.Through the teaching in wards and at the outpatient service,the students mastered the requirements in the syllabus of dermatology and venereology and achieved good results.