Object To study the chemical constituents of whole plant of Callicarpa pedunculata R Brown Methods Various chromatographic techniques were employed for isolation and purification of its constituents; and their structures were identified by spectral analysis (IR, MS, 1HNMR, 13 CNMR) and chemical evidences Results Ten compounds were obtained from the chloroform extract of the whole plant Among them four were diterpenoids which were identified as: 14 ?-hydroxy-7, 15-isopimaradien-18-oic acid (Ⅰ), 16 ?, 17-dihydroxy-3-oxo-phyllocladanetriol (Ⅱ), 8, 11, 13, 15-abietetraen-18-oic acid (Ⅲ), and 6 ?-hydroxy nidorellol (Ⅳ); 2 flavonoids: 5-hydroxy-3, 4′, 7-trimethoxy-flavone (Ⅴ), and 3, 5-dimethyl kaempherol (Ⅵ); and the others were ursolic acid (Ⅶ), myoinositol (Ⅷ), ?-sitosterol (Ⅸ), and ?-amyrin (Ⅹ) Conclusion Compounds Ⅰ, Ⅱ, Ⅲ, Ⅳ were found in this plant for the first time and it was of interest to note that they were of different diterpenoid skeletons The content of ursolic acid attained to a high level up to 1%, which may be exploited as a natural resource for medical purposes

Objective To evaluate the effect of gamma irradiation on the proliferation,differentiation,and mineralization of murine osteoblastic cells,and to investigate the related molecular mechanism.Methods Osteoblastic cells were irradiated by different doses (0,0.5,1.0,2.0,5.0 Gy)of 137Cs γ-rays.Cell morphology was observed with a microscopy,cell viability was analyzed by MTT assay,and ALP activity was analyzed by the methods of enzyme histochemistry and PNPP.Meanwhile,gene expressions of ALP,osteocalcin (OC),collagen Ⅰ,osteoprotegerin (OPG) and receptor activator of nuclear factor-kB ligand (RANKL) were measured by semi-quantified RT-PCR.Results Cell viability decreased with the radiation doses over 1.0 Gy ( t =6.197 - 18.677,P ＜ 0.05 ).After radiation with a dose over 2.0 Gy,the cell number and the junctions of cell protrusions decreased,the cells had low refractivity and the activity and mineralization ability of ALP were also inhibited ( t =2.790 -2l.374,P ＜0.05).In addition,the expressions of ALP and OC mRNA were down-regulated significantly (t =3.563 -16.508,P ＜ 0.05) when the radiation dose was higher than 0.5 Gy,and the expressions of OPG,OPG/RANKL mRNA were down-regulated ( t =12.942,4.954,P ＜ 0.05 ) at 5 Gy.But the expressions of collagen Ⅰ and RANKL mRNA were not affected by irradiation.Conclusions The osteoblastic cells were significantly influenced by γ-irradiation,including morphological changes,inhibition of cell proliferation,differentiation and mineralization ability. Meanwhile,mRNA expressions of ALP and OC were downregulated.OPG/RANKL may be a main pathway of osteoblastic cell damage under high dose radiation.

Objective To explore the feasibility of using glycophorin A somatic mutation in peripheral erythrocytes,in order to evaluate the cancer risk of occupational medical exposure to ionizing radiation.Methods Totally 336 medical radiation workers were recruited as three groups (general radiation group,computer tomography group,intervention and radiation treatment group) and 112 healthy adults were selected as control by using stratified random cluster sampling method,where 176 medicalradiation workers and 58 health controls had a MN-heterozygous type.The erythrocytes were fixed and bound with fluorescent-labeled monoclonal antibody,and the glycophorin A somatic mutation frequency was assayed by a modified BR6-1W1 method using a FACScan flow cytometer.The individual susceptibility to radiation was investigated using micronuclei test and 3-Aminobenzamide index test.Results The GPA somatic mutation frequency of medical-radiation workers was significantly higher than that of healthy control ( t =2.29 - 11.48,P ＜ 0.05 ).In particular,the NO GPA aberration frequency of interventional radiology workers was much higher than that of the general medical diagnostic workers (t =2.01,P ＜ 0.05).In addition,the NO GPA variant frequency changed significantly with the years of radiation service,cumulative doses,and 3AB index.However,the NN GPA variant frequency was only associated with the years of radiation service,and no significant correlations were found between NN GPA variant frequency and cumulative dose of radiation exposure or 3AB index. Conclusions GPA mutation frequency,especially NO GPA mutation frequency could be used as a sensitive biomarker to predict the DNA damage and individual susceptibility for the population exposed to professional low-dose ionizing radiation.

Thirty seven patients with congestive heart failure (CHF) were divided into cilazapril group (n=19) and general treatment group (n=18). Serum levels of interleukin-6 (IL-6) ,interleukin-8 (IL-8) , interleukin-10(IL-10) and tumor necrosis factor-α(TNF-α) , left ventricular ejection fraction (LVEF) ,left ventricular end-diastolic diameter (LVEDD), cardiac output (CO) and fractional shortening (FS) were measured before and after treatment. Serum levels of cytokines were also measured in 40 healthy individuals (control group). Results: The serum levels of IL-6, IL-8, IL-10 and TNF-α in CHF patients were significantly higher than those in the control group ( all P<0.01 ) ; After treatment, the serum IL-6, IL-8 and TNF-α were significantly decreased (P<0.01 ,P<0.05 ) in the cilazapril group. The LVEF, FS, CO were significantly increased in the Cilazapril group ( P<0.01 ) ; And the serum levels of IL-6 were significantly decreased in the cilazapril group as compared with the general treatment group ( P<0.05 ), however, after treatment, the EF, LVEF, FS and CO had no statistical differences in the cilazapril group as compared with the general treatment group. In the control group only LVEF and FS improved(P<0.01) ; Cytokine levels showed no changes. It suggests that cilazapril can reduce the serum levels of pro-inflammatory cytokines, increased the serum levels of anti-inflammatory cytokine, protect and improved cardiac function in the patients with congestive heart failure.

Objective To measure the expression of protein kinase D1 (PKD1),tyr463-phosphorylaed PKD1 (pPKD1-tyr463) and ser916-phos-phorylaed PKD1 (pPKD1-ser916) in squamous cell carcinoma (SCC),Bowen's disease (BD) and actinic keratosis (AK),and to explore their significance.Methods Fresh tissue samples were resected from lesions of patients with SCC (SCC group),BD (BD group) and AK (AK group),as well as from normal skin of healthy human controls (control group),and each group had a sample size of 10.Real-time RT-PCR was performed to measure the mRNA expression of protein kinase D1 gene (PRKD1),and Western blot analysis to determine the protein expression of PKD1,pPKD1-tyr463 and pPKD1-ser916.In addition,immunohistochemical study was conducted to determine the expression of PKD1,pPKD1-tyr463 and pPKD1-ser916 in another 50 paraffin-embedded skin samples of SCC,20 samples of BD,20 samples of AK and 10 normal skin samples.Results PRKD1 mRNA expression significantly differed among the control group (0.64 ± 0.09),SCC group (5.37 ± 1.06),BD group (2.69 ± 0.72) and AK group (2.43 ± 0.46) (F =21.37,P ＜ 0.05),and was significantly higher in the SCC,BD and AK groups than that in the control group (P ＜ 0.05),as well as in the SCC group than that in the AK and BD groups (both P ＜ 0.05).However,no significant difference in the PRKD1 mRNA expression was observed between the BD group and AK group (P ＞ 0.05).Immunohistochemical study showed that the total PKD1 protein and pPKD1-tyr463 in the SCC and BD groups were mainly expressed in the cytoplasm and cell membrane of spinous layer cells and atypical cells,and their expression rates were significantly higher than those in the AK group and control group (all P ＜ 0.01).The pPKD1-ser916 was only slightly expressed in some cancer nests of well-differentiated SCC tissues,but not in poorly-differentiated SCC,AK,BD tissues and normal skin tissues.In the SCC group,the expression rate of PKD1 increased with the increase of the pathological grade of SCC,and the PKD1 expression was positively correlated with pPKD1-tyr463 expression (rcc =0.479,P ＜ 0.05).Western blot results were consistent with immunohistochemical findings.Conclusion PKD1 and pPKD1-tyr463 may be involved in the development and differentiation of skin tumors derived from stratified squamous epithelium,and PKD1 may exert promotive effects on the formation of cutaneous SCC by activating the Tyr463 phosphorylation site.

Objective To discuss the pulmonary function change in infants with human bocavirus (HBoV) pneumonia or mycoplasma pneumonia (MP) and its clinical signiifcance. Methods One hundred and forty infants under 3 years old who were admitted due to pulmonary infection were recruited from January, 2013 to October, 2013. Among them, HBoV-DNA was detected in 64 cases, and MP-DNA was detected in 76 cases. Thirty eight normal age-matched infants were selected as controls. The shape of tidal breathing lfow-volume loops (TBFVLs) and change of every index were measured with a Pulmonary Testing System (Jaeger MasterScope). Results The ratio of time to peak tidal expiratory lfow (PTEF) to total expiratory time (TPTEF/TE), the ratio of VPTEF to expiratory volume (VPTEF/VE), tidal expiratory lfow at 25%of the remaining tidal volume (TEF25%) and the ratio of TEF25%to PTEF (25/PF) were signiifcantly decreased in infants with HBoV and MP infection as compared with healthy infants (P<0.05). However, there was no signiifcant difference of the above indices between infants with HBoV and MP infection (P>0.05). The shape of TBFVLs in infants with HBoV and MP pneumonia was changed and characterized by left-shifted PTEF and trough-like concave in descending limb. The PTEF was decreased in infants with pulmonary infection. Conclusions HBoV or MP infection results in impaired pulmonary function with manifestations of obstruction in small airway. The shape of TBFVLs in infants with HBoV and MP pneumonia is characterized by left-shifted PTEF and trough-like concave in descending limb.

Objective To study the Diagnosis and treatment of thyroglossal duct cyst or fistula. Methods The clinical data of 67 patients with thyroglossal duct cyst or fistula treated by surgery were analyzed retrospectively. Results The diagnosis rate preoperatively was 92.5% and the accuracy diagnosis rate by B type ultrasonography was 96.1%; all cases were treated by operation, of them, 14 cases underwent simple cyst resection, 12 with resection of partial periosteum of hyoid and 41 with resection of middle segment of hyoid( Sistrunk's operation ). 54 cases were followed up, recurrence in cases with cyst was 4 of 43 (9.3%) and recurrence in cases with fistula was 2 of 11(18.2%)respectively. Conclusion Ultrasonography shows certain characteristics and clinical value to diagnose and distinguish thyroglossal duct cyst. Postoperative recurrence is due to the selection of operation procedures and control of infection of focus. The Sistrunk's operation properly performed previou to cyst infection should reduce recurrence rate markedly.

Objective To evaluate the effect of aminolevulinic acid-based photodynamic therapy (ALA-PDT) on the expression of protein kinase D1 (PKD1) in a cutaneous squamous cell carcinoma cell line A431,and to explore the mechanism underlying ALA-PDT-induced apoptosis of A431 ceils.Methods A431 cells were cultured in vitro,and cell counting kit-8 (CCK-8) assay was performed to select the optimal combination of ALA concentration and PDT dose with the strongest proliferation inhibitory effect.A431 ceils at exponential growth phase were randomly divided into 4 groups:control group receiving no treatment,ALA group treated with ALA solution alone,PDT group treated with PDT alone,and ALA-PDT group treated firstly with ALA solution and then with PDT.After 12-,24-,36-and 48-hour additional culture,CCK-8 assay was conducted to evaluate the cellular proliferation inhibition,and the apoptosis rate at the time point of the strongest proliferation inhibitory effect was measured by flow cytometry.RT-PCR was performed to determine the expression of protein kinase D1 gene (PRKD1) in A431 cells at different time points after the ALA-PDT treatment,and Western blot analysis to measure protein expression of PKD 1 and its phosphorylation at Tyr463 (pTyr463) and Ser916 (pSer916) in A431 cells.Results The combination of ALA at the concentration of 1.5 mmol/L with PDT at an irradiation dose of 2 J/cm2 was optimal due to its strongest proliferation inhibitory effect.After 12-,24-,36-and 48-hour additional culture,there were significant differences in the proliferation inhibition rate among the 4 groups (F =39.56,P ＜ 0.05).At 24 hours after the treatment,the ALA-PDT group showed significantly higher proliferation inhibition rate (46.26％ ± 1.25％) compared with the ALA group (14.65％ ± 0.33％,P ＜ 0.05),PDT group (14.96％ ± 0.68％,P ＜ 0.05) and control group (11.98％ ± 0.32％,P ＜ 0.05),as well as compared with that at 12 hours (P ＜ 0.05).At 24 hours after the treatment,the apoptosis rate significantly differed among the 4 groups (F =16.32,P ＜ 0.05),and the ALA-PDT group showed a significantly higher apoptosis rate (41.92％ ± 3.23％) compared with the control group (4.67％ ± 0.88％,P ＜ 0.05),ALA group (7.02％ ± 1.52％,P ＜ 0.05) and PDT group (8.37％ ± 0.59％,P ＜ 0.05).At 0,6,12,24,36 and 48 hours after the treatment,there were significant differences in the mRNA expression of PRKD 1 among the 4 groups (F =22.24,P ＜ 0.05),and the mRNA expression of PRKD1 at 24 hours was significantly lower than that at 0,6,12 hours (all P ＜ 0.05),but was not significantly different from that at 36 and 48 hours (both P ＞ 0.05).No significant difference in the Ser916-phosphorylated PKD1 expression was found among the 4 groups (F =1.53,P ＞ 0.05),while there were significant differences in the expression of PKD1 and Tyr463-phosphorylated PKD 1 among the 4 groups (F =10.04,8.27,both P ＜ 0.05).Additionally,the ALA-PDT group showed significantly lower expression of PKD 1 and Tyr463-phosphorylated PKD 1 compared with the control group,ALA group and PDT group (all P ＜ 0.05).Conclusion PKD1 may be involved in the photochemical process of A431 cell apoptosis induced by ALA-PDT,and may promote the occurrence of squamous cell carcinoma by Tyr463 phosphorylation.

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 neurodevelopmental toxicity of ACR by studying the expression of DCX and GAP-43 in the hippocampal dentate gyrus of rats after maternal exposure to acrylamide. Methods Pregnant rats were randomly divided into low-dose ACR(4.5 mg/kg),medium-dose(9 mg/kg),high dose groups(18 mg/kg)and the control group(0 mg/kg),8 in each group,and were exposed to toxicant from gestation-al day 15 to postnatal day 13. All rats and their pups were killed on postnatal day 14. ABC immunohistochemistry was used to detect the expression of GFAP in the hippocampus of mother rats and offspring. Results Compared with the control group,the expression of DCX and GAP-43 in hippocampus dentate gyrus of the pregnant rats in middle and high dose groups was significantly decreased(P < 0.05). Conclusion ACR may interfere with the growth and development of neurons by reducing the expression of DCX and GAP-43.