An 82-year-old Chinese woman presented with skin eruptions on the thigh and abdomen accompanied by intermittent fever of unknown origin for more than 2 months.No hepatosplenomegaly,lymphadenopathy or neurological abnormality was found with physical examination.There were irregular,tender,indurated,dark-erythematous plaques on bilateral thigh and lower abdomen.along with nonpitting edema and peau d'orange appearance.A significant decrease was observed in the count of white blood cells,red blood cells and platelets,but the serum level of lactate dehydrogenase was elevated.Tumor aspiration and the first pathology yielded no confirmed diagnosis,and the patient had ever been diagnosed with chronic lymphangitis,allergic cutaneous vasculitis and fever of unknown origin in other hospitals.Antibiotic therapy leaded to no improvement,and the lesions gradually spread from the migh to lower abdomen.The second histopathology revealed the presence of atypical lymphoid cells with hyperchromatic nuclei and irregular morphology in the lumens of small blood vessels in subcutaneous tissue.Immunohistochemically,the atypical lymphoid cells were positive for lymphocytotoxic antibody (LCA),CD20,CD790t and bcl-2,but negative for bcl-6,CD10,CD3,CD45RO,CD30,EMA,AEI/3 or CK and vascular endothelial cells were positive for CD34.The diagnosis of intravascular large B-cell lymphoma was made based on the hisstopathological features and immunohistochemical findings.The patient died in two months.

ObjectiveTo discuss the relationships among spastic paralysis and motor function, balance function and other clinical variables after stroke.MethodsAssessed the clinical spastic index of hemiplegic lower limbs with CSI scale, motor function with Fugl-Meyer Assessment, and balance function with Berg Equilibrium Scale, respectively. And then analyzed the relationship between clinical spasm index with motor function, balance function of the hemiplegic lower limbs with Pearson correlation analysis. Finally, ascertained which was the most important factor affecting the clinical spasm index of spastic lower limbs with stepwise regression analysis.ResultsThere is negative correlation between clinical spasm index with motor function and balance function, respectively. Tendon reflex, muscular tension and clonus are the deciding factors to motor function(P<0.05) and balance function(P<0.05) of the hemiplegic lower limbs.ConclusionThe clinical spasm index of hemiplegic limbs can affect the recovery of motor function and balance function significantly. So, assessing and improving the clinical spasm index of paralytic lower limbs can optimize the rehabilitation program to stroke patients.

Objective: To investigate the contamination status and assess the potential consumption risk of Listeria monocytogenes ( L. monocytogenes ) in cooked meat products in bulk in Zhejiang Province, so as to provide strategy for food safety supervision and management. Methods: A total of 2 320 cooked meat products were sampled from eleven cities in Zhejiang Province during 2018-2020. The detection of L. monocytogenes was carried out in accordance with the national standard GB/T 4789.30-2016. Risk Ranger software was used for the semi-quantitative risk assessment on the whole population and pregnant women. Results: The total detection rate of L. monocytogenes in cooked meat products in bulk in Zhejiang Province was 2.97% ( 69/2 320 ). The detection rates in stewed, smoked/roasted, fried, dried products and others were 3.85%, 1.81%, 0.59%, 0% and 0.94%, which were significantly different ( P<0.05 ). There were 28 positive samples in 1 069 samples collected in 2020, with the concentration ranging from 5 to 590 CFU/g and averaging 6.8 CFU/g. The estimated number of listeriosis cases each year caused by consumption of cooked meat products in bulk was 131 in the whole population with a risk score of 42, and 1.44 in pregnant women with a risk score of 54. The risk coefficient could reduce to approximate zero after sufficient heating before intake. Conclusion The prevalence of L. monocytogenes in cooked meat products in bulk in Zhejiang Province during 2018-2020 poses a potential risk in food safety. Pregnant women should avoid eating.

Objective: To establish a rapid GeXP-based multiplex reverse transcription-PCR assay (GeXP assay) for simultaneous detection of 5 subtypes of diarrheogenic Escherichia coli. Methods: Specific primers were designed according to reserved sequences of 12 virulence genes in enterotoxigenic E. coli (ETEC), enterinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC) and enterohemorrhagic E. coli (ETEC), and PCR amplification was performed with a single pair of primers to validate the specificity of PCR assay with a single template and a single pair of primers. The specificity of the GeXP assay was evaluated with the genomic DNA of 5 subtypes of diarrheogenic E. coli as the template in a mixture of 12 pairs of primers, and the sensitivity of the GeXP assay was evaluated with the mixed suspensions of 5 subtypes of diarrheogenic E. coli at concentrations of 106, 105, 104 and 103 CFU/mL as the template. Foods purchased from supermarkets and agricultural retail markets were prepared into 34 spiked samples, and 5 subtypes of diarrheogenic E. coli were detected using the GeXP assay and compared with the fluorescent real-time multiple PCR assay. Results: The sizes of sth, pic, bfpB, astA, lt, escV, aggR, stx1, uidA, invE, stx2 and stp genes amplification products were consistent with expected sizes using a single template and a single pair of primers, with a fluorescent signal intensity of more than 25 000 A.U. The sizes of the GeXP assay amplification products of 12 virulence genes in 5 subtypes of diarrheogenic E. coli were consistent with expected sizes, with a high specificity. If the concentration of the mixed suspensions of 5 subtypes of diarrheogenic E. coli was 103 CFU/mL, the GeXP assay was effective for simultaneous detection of 12 virulence genes, with a high fluorescent signal intensity, consistent repeated detection results and a less than 10% coefficient of variation. The GeXP assay detected 3 ETEC isolates, 12 EAEC isolates, one EIEC isolate, one EPEC isolate and one EHEC isolate among the 34 spiked samples, which was in agreement with the detection of 5 subtypes of diarrheogenic E. coli with commercial fluorescent real-time multiple PCR assay kits. Conclusions A GeXP assay has been successfully established for simultaneous detection of 12 virulence genes in diarrheogenic E. coli, which is effective for clinical differential diagnosis and epidemiological surveys of diarrheogenic E. coli.

OBJECTIVE To understand nosocomial candidal infection among tuberculosis patients and its drug(sensitivity).(METHODS) Totally 6280 clinical samples from Jan 2002 to Dec 2004 were cultured;and part of(positive) ones were tested their drug sensitivity to FC,FLC,KET and ITC.RESULTS From all cultured samples 2767 Candida strains(positive rate was 44.06%) were isolated.From them C.albicans,C.glabrata,C.tropicalis,C.krusei and other Candida were 2006, 344,206,95 and 116 strains,their positive rate was 31.94%,(5.48%),3.28%, 1.51% and 1.85%,respectively.Their drug sensitivity rate to four drugs were as follows: to FC was 93.73%,to TTC 62.76%,to FLC 51.03% and to KET was 47.59%. CONCLUSIONS The condition of(tuberculosis) patient would become serious due to higher dual infection rate by Candida after too long anti-(tuberculosis) treatment.Among the four kinds of drugs the FC has the best anti-candidal activity.

Objective:To investigate the clinical efficacy of postoperative radiotherapy combined with compound matrine injection on the patients with breast cancer.Methods:Eighty-three patients with breast cancer received breast-conserving and sentinel lymph node biopsy before the study who were admitted in our hospital were divided into the control group and observation group.Forty-two patients in control group were treated with postoperative radiotherapy,and forty-one patients in observation group were received postoperative radiotherapy combined with compound matrine injection.The CA125,CA153,CA724,levels of CD3+,CD4+,CD8+,CD4+/CD8+,NK,and adverse reaction in the two groups were detected and compared.Results:After treatment,the CA125,CA153,and CA724 in both groups were obviously decreased (P＜0.05),which were significantly lower in the observation group than those of the control group (P＜0.05).CD3+,CD4+,CD8+,CD4+/CD8+,and NK cell in the observation group showed remarkable improvement (P＜0.05) compared with those before treatment.And CD3+,CD4+,CD4+/CD8+,and NK cell of the observation group were significantly higher than those of the control group,while CD8+ was much lower (P＜0.05).During the treatment period,the incidence of skin reaction over grade Ⅱ,and myelosuppression in observation group were much lower than those of control group (P＜0.05).Conclusion:Postoperative radiotherapy combined with compound matrine injection had good ability in enhancing the cellular immune function and reducing adverse reaction on the patients with breast cancer.

Objective To observe the effects of repeated Botulinum toxin type A (BTX-A) injection on lower limb spasticity after stroke.Methods 180 cases with lower limb spasticity after stroke were divided into the treatment group (n=90) and the control group (n=90). The treatment group was treated with BTX-A injection twice in the spastic muscles at interval of 3~6 months, while both the treatment group and the control group accepted the rehabilitation based on the neurodevelopmental therapy. They were assessed with modified Ashworth Scale (MAS), Fugl-Meyer Lower Limb Assessment (FMAL), Berg Balance Scale (BBS), modified Barthel Index (MBI) before each injection, and 3 d, 7 d, 1 month, 3 months after each injection or the same time for the controls. Results There was significant difference in scores of MAS, FMAL, BBS, MBI for the treatment group among before and 3 d, 7 d, 1 month after each injection (P<0.05), but not significant between 2 injections (P<0.05). There was significant difference in scores of all the assessment between the treatment and control group at the same time (P<0.01). Conclusion Repeated intramuscular injection of BTX-A can reduce the spasticity of lower limb after stroke.

Matteuccia struthiopteris is a nature plant, which contains a lot of potential active components. In the present study, we investigated the effect of polysaccharides extracted from Matteuccia struthiopteris on lupus-like syndrome induced by Campylobacter jejuni CJ-S131 in BALB/c mice. Mice were randomly divided into normal, model control, SLE model (vehicle treated), Matteuccia struthiopteris polysaccharides treated (30 and 15 mg x kg(-1)) groups and prednisone 5 mg x kg(-1) treated groups. The effect of Matteuccia struthiopteris polysaccharides (Ms) on weight and organ index of BALB/c mice was detected. Autoantibodies and total IgG production were measured by enzyme linked immunosorbent assay. Proteinuria was measured and kidneys were examined by light microscopy. Compared with SLE model group, treatment with Matteuccia struthiopteris polysaccharides 30 and 15 mg x kg(-1) reduced weight loss and Matteuccia struthiopteris polysaccharides 15 mg x kg(-1) reduced spleen swelling (P < 0.05). The increased production of autoantibodies and total immunoglobulin G (IgG) were also significantly inhibited. Matteuccia struthiopteris polysaccharides protected kidney against glomerular injury in BALB/c mice with reduced immunoglobulin deposition and lowered proteinuria (P < 0.01). Matteuccia struthiopteris polysaccharides had a protective effect on lupus-like syndrome induced by CJ-S131 in BALB/c mice.

Objective To detect and explore the expression and clinical significance of dual specificity tyrosine phosphorylation regulated kinase1b (Dyrk1b) in the specimens and cells of cervical lesions. Methods (1)All the data were collected from 75 patients with cervical cancer and 52 cases with squamous intraepithelial lesion(SIL)admitted in the First Affiliated Hospital of Dalian Medical College during Jan. 2011 to Dec. 2013 and confirmed by pathological examination, included 60 cases of stageⅠand 15 cases of stageⅡ, 12 cases with low-grade squamous intraepithelial lesion(LSIL)and 40 cases with high-grade squamous intraepithelial lesion(HSIL). While, 28 cases with chronic cervicitis were chosen as the control group. The protein expression of Dyrk1b was detected by immunohistochemistry among the four groups.(2)The expression of Dyrk1b in HeLa and SiHa cells were detected by western blot method and the expression of Dyrk1b protein were also detected after treatment of AZ191 (5, 10 μmol/L) for 48 hours in HeLa and SiHa cells.(3)The cellular survival and proliferation of HeLa and SiHa cells treated by different concentrations of AZ191(2.5, 5, 10, 25, 50, 100 μmol/L)for 48 hours were detected by methyl thiazolyl tetrazolium (MTT) assay.(4)The rate of apoptosis of HeLa and SiHa cells was detected by flowcytometry after treatment of AZ191 (5, 10μmol/L) for 48 hours. Results (1)The positive rates of Dyrk1b protein in chronic cervicitis, LSIL, HSIL and cervical squamous cancer by immunohistochemistry were 11%(3/28), 1/12, 42%(17/40)and 71%(53/75), respectively. The expression of Dyrk1b in cervical squamous cancer and HISL were higher than those in LSIL and chronic cervicitis (P<0.01), there were significant difference between cervical squamous cancer and HSIL, or between HSIL and LSIL(all P<0.05), while there were not significant difference between LSIL and chronic cervicitis(P>0.05). Expression of Dyrk1b was correlated with stromal invasion depth of cervical cancer (P<0.05), but not with age, clinical stage, lymph node metastasis, and serum squamous cell carcinom antigen(SCC-Ag)levels (all P>0.05). (2) Dyrk1b protein was expressed in different levels in HeLa and SiHa cells, and the expression of Dyrk1b was decreased gradually as the increased of the concentration of AZ191 in both HeLa and SiHa cells by treatment of AZ191 for 48 hours. (3) Different concentration of AZ191 treated on cervical cancer cells could inhibit the cellular proliferation and induce cell apoptosis in a concentration-dependent manner(P<0.01), concomitant to the decreased cell survival rate. The apoptosis rate of HeLa and SiHa were increased significantly after 10μmol/L AZ191-treatment for 48 hours, but no any difference induced by 5 μmol/L AZ191-treatment compared to control group. Also,there was no any difference between Hela and SiHa cells in either inhibitory effect or apoptosis rate induced by AZ191. Conclusions Dyrk1b is over-expressed in either specimens or cells of cervical cancer. The expression of Dyrk1b protein in cervical lesions is increased as the progression of disease. Dyrk1b inhibitor AZ191 could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner in cervical cancer cells.

Objective: To establish real-time recombinase polymerase amplification（RPA）for the rapid detection of Vibrio parahaemolyticus（VP）. Methods: An exo probe and primers were designed according to the conserved sequence of thermolabile hemolysin（tlh）gene of VP and then RPA for detection of VP was established. The sensitivity of the assay was evaluated by detecting different concentration of VP;the specificity was evaluated by detecting different bacteria;the stability was evaluated by repeat trials;the application effect was evaluated by detecting food samples which were simultaneously tested with traditional culture method according to GB 4798.7-2013 Detection of VP. Results: A real-time RPA was established to complete VP amplification within 20 min at a constant temperature of 39 ℃. The analytical sensitivity of the assay was five pg per reaction and no cross-reactivity with other pathogenic bacteria observed. The RPA detection results with different concentration of VP and E. coli DNA templates at three time points were consistent. The detection results of 51 food samples by RPA were the same as those by traditional culture method. Conclusion The established real-time RPA can qualitatively detect VP,with simple operation and interpretation of results,which is suitable for rapid detection of VP in public health emergencies and food safety supervision.