Objective To observe the effect of δ-opioid receptor on proliferation and migration of human epidermal stem cells (hESCs) in vitro so as to offer treatment theory for skin injury.Methods hESCs from fresh foreskin tissues of normal young volunteers were isolated and cultured by enzyme digestion and differential adherence technique.Immunofluorescent staining was used to determine expression of integrin β1 and cytokeratin 19 (CK19) and flow cytometry was used for cell count.Second generation of cells were cultured for 5 consecutive days with keratinocyte serum-free medium (K-SFM) supplemented with 1 nmol/L (D-Ala2,K-Leu5)-enkephalin in Group A,with K-SFM supplemented with 1 nmol/L naltrindole and 1 nmol/L (D-Ala2,K-Leu5)-enkephalin in Group B,and with isolated K-SFM in Group C.Cellular division and proliferation were detected by MTT method.An in vitro 100 μm scratch-wound model was created on the confluent monolayer cells at 24 hours of incubation.Cells migrating from the wound margin were determined by inverted phase contrast microscope at 24,48,72,and 96 hours after wound formation,while wound closure rate was calculated at 72 hours.Results Primary cultured hESCs presented cobblestone-like shape after adherence growth,Immunofluorescence staining showed positive results for integrin β1 and CK19 and cell purity reached 95.6％.Moreover,MTT findings revealed proliferation of hESCs enhanced significantly in Group A,but lowered in Group B as compared to Group C (P ＜ 0.05).hESCs migrated from the wound margin in all groups at 24 hours.However,more migrated cells were seen in Group A than in Group C and less in Group B than in Group C.Rate of wound closure was (89.5 ±0.7)％ in Group A,(76.1 ±0.3)％ in Group B,and (81.1 ±0.6)％ in Group C at 72 hours,indicating significant differences among groups (P ＜ 0.05).Conclusion Activation of δ-opioid receptor promotes the proliferation and migration of hESCs in vitro and may be implicated in wound healing.

BACKGROUND:Macrophage migration inhibitory factor is involved in the process of a variety of diseases, and plays a very important role in the tumor, autoimmune diseases, inflammation, angiogenesis, fibrotic diseases and so on. These biological characteristics are similar to keloids. OBJECTIVE: To compare the distribution and number of macrophage migration inhibitory factor in normal skin, hypertrophic scar and keloid. METHODS: We colected 40 clinical pathological scar specimens after surgery, including 20 hypertrophic scars and 20 keloids. Another 10 samples of the normal skin were used as control group. Hematoxylin-eosin staining and immunohistochemistry staining were performed to test the expression of macrophage migration inhibitory factor in pathological scars and normal skin. RESULTS AND CONCLUSION:Macrophage migration inhibitory factor was positively expressed in the normal skin, hypertrophic scar and keloid, and the expression of macrophage migration inhibitory factor in keloid was significantly higher than that in hypertrophic scar and normal skin (P < 0.01). It means that the abnormal infiltration of macrophage migration inhibitory factor may be associated with the formation of keloid.

Objective To review the clinical characteristics of paraganglioma of the urinary bladder with 5 cases reprt.Methods The clinical data of 5 patients (2 males and 3 females,age 23-68 years)with paraganglioma of the urinary bladder were reported.Two cases were diagnosed by ultrasonography during health examination,1 case was found by hematuria,1 had difficulty of voiding,and 1 presented with palpitation,chest discomfort while urination.Two cases were clinical diagnosed as bladder paraganglioma,1case urachal carcinoma,and 2 cases bladder tumor.Cystoscopy showed a protruding tumor within the bladder or bladder had compressed changes.One case of tumor located in the triangle,1 in the posterior wall,1in the top,2 in the anterior wall.Three cases of biopsy were negative.Three cases of preoperative endocrine examinations showed norepinephrine significantly higher.Results Laparoscopic partial cystectomy was performed in 3 cases,open partial cystectomy in 1 case,and transurethral resection of bladder tumor (TURBT) in 1 case.One case had the complication of stress cardiomyopathy during TURBT,3 cases found intraoperative hypertensive crisis.The bleeding volume was 20-800 ml (average 126 ml),and I case received blood transfusion.During the follow-up period for 3-48 months,the blood pressure was normal,and no recurrence was found.Conclusions Bladder paraganglioma is uncommon and easily misdiagnosed.For the patients with bladder tumor,accompanied by changes in blood pressure,palpitations during urination should be highly suspicious of bladder paraganglioma.Partial cystectomy is the main treatment method.

Objective: To construct an evaluation index system of oral health literacy for children's parents, so as to provide an index system for the evaluation of oral health literacy for children's parents in China. Methods: The evaluation index system of oral health literacy for children's parents was designed based on literature review and semi-structured interview. Experts from oral prevention and pediatric oral medicine were invited to participate in two-round Delphi consultations. The indicators were scored and screened according to the importance, and the weight determined using analytic hierarchy process. The effectiveness of the consultation was evaluated by positive coefficient, authority coefficient and coordination coefficient. Results: Twenty-four experts participated in the consultation, including 6 males and 18 females. There were 21 experts with a master degree, 3 experts with a doctor degree, and 20 experts with vice senior professional titles and above. The recovery rates of the two rounds of consultations were 96.00% and 100.00%, the authority coefficients were 0.866 and 0.917, the Kendall's coefficient of concordance were 0.120 and 0.156 (both P<0.05), and the coefficient of variation was 0.15-0.38 and 0.03-0.17, respectively. The final evaluation index system included 3 primary indicators, 11 secondary indicators and 40 tertiary indicators. The primary indicators were basic knowledge and concepts related to oral health, promoting lifestyle and behaviors related to oral health, and maintaining basic skills related to oral health. Conclusion The evaluation index system of oral health literacy for children's parents has been established in this study and needs to be further applied and evaluated.

Objective: To investigate the epidemiological characteristics of human brucellosis in Jiaxing City from 2010 to 2021, so as to provide insights into the development of the brucellosis control strategy. Methods: The epidemiological and clinical data of brucellosis patients and epidemiological data of brucellosis outbreaks in Jiaxing City from 2010 to 2021 were collected from Chinese Disease Control and Prevention Information System, and the epidemiological features and outbreaks of brucellosis were analyzed descriptively. Results: Totally 160 brucellosis patients were reported in Jiaxing City from 2010 to 2021, and the incidence of brucellosis appeared a tendency towards a rise (χ2trend=28.564, P=0.002), with annual mean incidence of 0.29/105. No deaths due to brucellosis occurred in Jiaxing City from 2010 to 2021. Brucellosis cases were reported each month, which were concentrated in the first and second quarters, and the greatest number was seen in May (27 cases, 16.88%). The brucellosis cases were predominantly reported in Tongxiang City (114 cases, 71.25%), and 75.00% were male (120 cases) and 70.63% were occupational populations (113 cases). The patients had a median (interquartile range) age of 57 (12) years at onset, and the median duration (interquartile range) from onset to definitive diagnosis was 18 (28) days. The clinical manifestations mainly included fever and weakness, and a total of 18 Brucella melitensis isolates and one B. bovis isolate were cultured. Conclusions The incidence of brucellosis was rising in Jiaxing City from 2010 to 2021. The brucellosis patients were predominantly reported in Tongxiang City in the first and second quarters, and young, middle-aged men and occupational populations were at a high risk of brucellosis.

Objective:To explore the evaluation of cytochrome P450 2C19 *2 (CYP2C19 *2) gene polymorphism and Helicobacter pylori (Hp) infection for clopidogrel efficacy in patients with coronary heart disease. Methods:The clinical data of 113 patients with coronary heart disease from February 2016 to March 2020 in Suzhou High-tech Zone People′s Hospital were retrospectively analyzed. The CYP2C19 *2 gene polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, the Hp infection was detected by 13C urea breath test. The patients were treated with clopidogrel, the effect after 4 weeks was evaluated. The receiver operating characteristic (ROC) curve was used to evaluate the CYP2C19 *2 gene polymorphism and Hp infection for evaluating clopidogrel effect in patients with coronary heart disease. Results:The CYP2C19 *2 genotype in patients with coronary heart disease conformed to Hardy-Weinberg balance ( χ2 = 0.33, P>0.05). Among 113 patients with coronary heart disease, Hp infection was in 27 cases, and Hp non-infection in 86 cases. Among Hp infection patients, CYP2C19 *2 gene GG was in 2 cases, GA in 6 cases, AA in 19 cases; among Hp non-infection patients, CYP2C19 *2 gene GG was in 23 cases, GA in 46 cases, AA in 17 cases, there was statistical difference in CYP2C19 *2 gene polymorphism between the two ( χ2 = 24.35, P<0.01). After clopidogrel treatment, effectiveness was in 79 cases, inefficiency in 34 cases. Among effectiveness patients, YP2C19 *2 gene GG was in 20 cases, GA in 43 cases, AA in 16 cases; among inefficiency patients, CYP2C19 *2 gene GG was in 5 cases, GA in 9 cases, AA in 20 cases, there was statistical difference in CYP2C19 *2 gene polymorphism between the two groups ( χ2 = 16.35, P<0.01). The rate of Hp infection in effectiveness patients was significantly lower than that in inefficiency patients: 12.66% (10/79) vs. 50.00% (17/34), and there was statistical difference ( χ2 = 18.23, P<0.05). ROC curve analysis result showed that the area under the curve of CYP2C19 *2 gene polymorphism combined with Hp infection for evaluating clopidogrel effect in patients with coronary heart disease was larger than CYP2C19 *2 gent GG, GA, AA and Hp infection alone evaluating (0.973 vs. 0.869, 0.679, 0.884 and 0.728) . Conclusions:The CYP2C19 *2 gene polymorphism is associated with Hp infection in patients with coronary heart disease, and the CYP2C19 *2 gene polymorphism combined with Hp infection has the evaluation value for the efficacy of clopidogrel.

T cells engineered with chimeric antigen receptor (CAR) have been successfully applied to treat advanced refractory B cell malignancy.However,many challenges remain in extending its application toward the treatment of solid tumors.The immunosuppressive nature of tumor microenvironment is considered one of the key factors limiting CAR-T efficacy.One negative regulator of T cell activity is lymphocyte activation gene-3 (LAG-3).We successfully generated LAG-3 knockout T and CAR-T cells with high efficiency using CRISPR-Cas9 mediated gene editing and found that the viability and immune phenotype were not dramatically changed during in vitro culture.LAG-3 knockout CAR-T cells displayed robust antigen-specific antitumor activity in cell culture and in murine xenograft model,which is comparable to standard CAR-T cells.Our study demonstrates an efficient approach to silence immune checkpoint in CAR-T cells via gene editing.

Objective: To perform an epidemiological survey of the first case with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in Pinghu City of Jiaxing City, Zhejiang Province on March 13, 2022, so as to provide insights into the management of coronavirus disease (COVID-19) epidemics. Methods: According to the requirements of the Protocol on Prevention and Control of COVID-19 (8th Edition), epidemiological investigations were performed among 39 cases with SARS-CoV-2 infections in Pinghu City from March 13 to 20, 2022. Cases' demographics, clinical symptoms, history of immunization and exposure were collected, and close contacts were identified. Pharyngeal swabs were sampled from infected cases for detection of SARS-CoV-2 nucleic acid and whole-genome sequencing, and the source of infection and transmission route were investigated. Results: The index case for this COVID-19 epidemic was an imported case from Shanghai Municipality, who infected 6 persons via aerosol transmission when playing in the badminton venue of Pinghu National Fitness Center on March 9; subsequently, one of these infected cases infected another 18 persons when playing in the badminton venue of Jiadian Village Resident's Fitness Center in Zhapu Township on March 12. Sixteen confirmed cases were reported, and all cases were mild; another 23 asymptomatic cases were diagnosed, with no death reported. This epidemic occurred from March 11 to 20, with 3 generations of spread and a median incubation period of 3 days. The SARS-CoV-2 infected cases had a median age of 33.5 (interquartile range, 12.0) years and included 36 cases with a history of COVID-19 vaccination. There were 16 cases with fever, cough, runny nose and sore throat, and 13 cases with imaging features of pneumonia. The effective reproductive number (Rt) of the COVID-19 epidemic was 7.73 at early stage, and was less than 1 since March 21. Whole-genome sequencing identified Omicron BA.2 variant among 33 cases, which had high homology with the index cases. Conclusion This epidemic was a cluster of COVID-19 caused by imported Omicron BA.2 variant infection from Shanghai Municipality, and the COVID-19 transmission was mainly caused by indoor aerosols.

Objective: To investigate the clinical features, diagnosis and treatment of osteomyelitis of the jaw caused by an actinomycotic infection and to provide a reference for clinical diagnosis and treatment. Methods: A case of osteomyelitis in the bilateral maxilla and the left zygomatic bone and arch caused by a mixed bacterial infection dominated by Actinomycetes was reviewed and analyzed in combination with the literature. Results : The patient had left upper posterior tooth pain with repeated left facial swelling for 7 months. The patient's left face was swollen before surgery, the left maxillary alveolar bone was necrotic, and the upper palate showed fistula discharge. A maxillofacial magnetic resonance imaging scan excluded tumors and other space-occupying lesions. According to CBCT images, the initial diagnoses were left infraorbital space infection and osteomyelitis of the bilateral maxillary, the left zygomatic bone, the left zygomatic arch and the lateral orbital wall. Necrosis of the left maxilla and the zygomatic bone was excised, the focus was cleared and the focal tooth was extracted under general anesthesia. Histopathological results confirmed osteomyelitis and actinomycotic infection. Anti-inflammatory therapy with penicillin sodium was given before surgery, and piperacillin sodium and tazobactam sodium, dexamethasone sodium phosphate, tranexamic acid and mecobalamine were given after surgery. The patients' 6-month follow-up results showed that the maxillofacial shape was basically symmetrical; no ulceration, pus or abnormal secretion was found in the skin or intraoral mucosa; and the surgical area showed good recovery. A review of the relevant literature showed that Actinomyces is an opportunistic pathogen, and factors such as trauma and dental infection have been implicated in the pathogenesis of osteomyelitis. In addition to surgery, antibiotics are used to treat the disease and multidisciplinary symptomatic treatment combined with supportive treatment is required to achieve a better prognostic effect. Conclusion Actinomycotic osteomyelitis occurring in the maxilla and the zygomatic bone is an extremely rare disease that can be diagnosed by clinical manifestations, bacteriological examination and biopsy. Appropriate and effective penicillin drugs should be given at the initial stage of treatment, more sensitive antibiotics should be selected according to the results of the drug sensitivity test, and the lesions should be surgically removed when the patient's condition improves. Active symptomatic and supportive treatment should be performed during the treatment period.

Objective To test the cardiac toxicity of new compound HMS-01 and evaluate the safety profile for clinical trials. Methods Manualpatch clamp method was used to measure human Ether-a-go-go-Related Gene (hERG) potassium channel currents with different concentrations of HMS-01. Cisapride was selected as the positive control drug. HMS-01 was diluted to the concentration of 0.3, 1, 3, 10 and 30 µmol/L and applied to the cells. The changes in electrical currents were recorded and the inhibition rate was calculated. Results At the highest concentration of 30µmol/L, the inhibitory rate of HMS-01 on hERG channel was less than 30%. There was no obvious inhibitory effect compared with cisapride. Conclusion Compared with the cisapride, HMS-01 has no obvious inhibitory effect on hERG channel and has no cardiotoxicity.