β?carbonic anhydrases(β?CAs)are ubiquitous metalloenzymes which active site contains a zinc ion(Zn2+),and they could catalyze the hydration of carbon dioxide to bicarbonate and protons efficiently and are involved in many biological pro?cesses,such as respiration,pH and CO2 homeostasis,biosynthetic reactions,virulence regulation and so on,and may play a critical role in the life activity of many organisms which contain these enzymes. β?CAs are widely distributed in fungi,bacteria, algae,plants and a small number of protozoan and metazoan except vertebrates. Therefore,as potential drug targets for design?ing and developing antibacterial and anti?parasitic drugs,β?CAs promise a broad application prospect. This paper focuses on the distribution,physiological function and the progress of researches on β?CAs in parasites and their vectors.

BACKGROUND:Neuronal calcium sensor-1 protein has a variety of different neuronal functions and has a high distribution in different areas of the brain. A single residue R102Q mutation in human neuronal calcium sensor-1 protein is demonstrated to be associated with autism disease. The experiment studies have reported that this R102Q mutant has essential conformation changes in local area of the neuronal calcium sensor-1. OBJECTIVE: To wel understand the specific reasons of the R102Q mutation of the neuronal calcium sensor-1 to the conformational dynamic changes. METHODS:Six independent extensive al-atom molecule dynamic simulations during 0-450 ns were conducted. RESULTS AND CONCLUSION: We have found that (1) there is no obvious recombination during the simulations between wild type and mutant type, but R102Q mutant alters the helix and makes the structure of the protein more stable; (2) R102Q mutation alters the salt bridges, reduces the flexibility of L2, and makes L3 extend in hydrophobic crevice. These results reveal that the helix plays an important role in the structural stability, and salt bridge is the important reason for the dynamic changes of neuronal calcium sensor-1 protein. This study may provide a structural insight into the function of protein deficiency associated with R102Q mutant.

Helicobacter pylori (H.pylori) infection is the most important pathogenic microorganisms associated with the development of chronic gastritis, peptic ulcer and gastric mucosa-associated lymphoid tissue lymphoma (MALT) in children. H.pylori infection may occur in early childhood and last a lifetime even directly affecting the health of adulthood unlessH.pylori infection was eradicated. This article aims to review the infection condition, infection pathway and immunological characteristics ofH.pylori and its relationship with extragastroduodenal diseases in children.

ObjectiveTo analyze the characteristics of severe trauma patients with acute kidney injury (AKI) receiving renal replacement therapy (RRT), in order to look for the risk factors of AKI and the opportune time for the initiation of RRT on prognosis.Methods A retrospective cohort study involving consecutive patients with severe trauma in emergency intensive care unit (ICU) in the Second Affiliated Hospital of Zhejiang University School of Medicine, from August 2011 to December 2014, was conducted. Inclusion criteria included age≥18 years, injury severity score (ISS)> 16, AKI receiving RRT, and the duration of hospital stay> 24 hours. The general data, the risk factors of AKI, the prognostic indicators, and the information of RRT were recorded. All patients were divided into two groups according to the prognosis, the time of onset of AKI and the initiation time of RRT. The independent risk factors for prognosis were screened by binary logistic regression analysis.Results Seventy-three patients were eligible for enrollment, including 48 deaths (65.8%); 49 patients suffered from AKI≤48 hours after trauma (early stage group), and in 24 patients it was longer than 48 hours (late stage group). In 55 patients RRT was routinely started (routine RRT group), 18 patients underwent RRT ahead of routine criteria decided by the judgment of the attending doctor (earlier RRT group). The main risk factors of RRT in traumatic patients with AKI were shock and sepsis, each accounted for 90.4% and 53.4%. Compared with survival group, in death group, the proportion of male patients was lower (70.8% vs. 100.0%,χ2 = 7.238,P = 0.007), acute physiology and chronic health evaluationⅡ (APACHEⅡ) scores were higher (23.7±5.1 vs. 14.4±3.7,t = 8.031,P< 0.001), Glasgow coma score (GCS) was lower [5.0 (3.0, 15.0) vs. 15.0 (8.0, 15.0),U = 320.000,P = 0.001], incidence of shock and sepsis was higher (97.9% vs. 76.0%,χ2 =6.755,P = 0.009; 64.6% vs. 32.0%,χ2 = 7.014,P = 0.008), the rate of use of contrast medium was lower (27.1% vs. 56.0%,χ2 = 5.898,P = 0.015), the time for the diagnosis of AKI post trauma was delayed [days: 2 (1, 5) vs. 2 (1, 2), U = 762.000,P = 0.049], the time for the initiation of RRT post trauma was later [days: 6.0 (3.0, 12.0) vs. 3.0 (2.0, 4.5), U = 868.500,P = 0.002], the recovery rate of renal function at discharge was lower (10.4% vs. 100.0%,χ2 = 54.497, P< 0.001). Compared with late stage group, in early stage group, the mortality was lower (55.1% vs. 87.5%,χ2 =7.509,P = 0.006), and the incidence of sepsis before AKI was also lower (38.8% vs. 83.3%,χ2 = 12.854,P< 0.001). Compared with routine RRT group, the recovery of renal function at discharge was better with a lower mortality rate in the earlier RRT group, but the difference was considered to be insignificant (55.6% vs. 36.4%,χ2 = 2.064,P = 0.151;50.0% vs. 70.9%,χ2 = 2.633,P = 0.105). Logistic regression analysis showed GCS [odds ratio (OR) = 0.852, 95%confidence interval (95%CI) = 0.747-0.972,P = 0.017], shock before AKI (OR = 85.350, 95%CI = 5.682-1 282.073, P = 0.001), and sepsis before AKI (OR = 11.499, 95%CI = 2.127 - 62.161,P = 0.005) were independent risk factors for the judgment of prognosis.Conclusions Shock and sepsis are the major risk factors of RRT in trauma patients with AKI. Shock, sepsis and traumatic brain injury are the independent risk factors of death. Perhaps early initiation of routine RRT cannot improve the outcome of the patients with posttraumatic renal insuficiency.

Objective:To observe expressions of vascular endothelial growth factor (VEGF ) and placental growth factor (PLGF) in patients with hypertensive disorder complicating pregnancy (HDCP) during early and mid pregnancy ,and pro‐vide basis for diagnosing HDCP .Methods:A total of 46 normal pregnant women (normal pregnancy group ) and 39 HDCP patients (HDCP group) were enrolled .The HDCP group was further divided into gestational hypertension group (n=26) , mild preeclampsia group (n=8) and severe preeclampsia group (n=5) .Enzyme linked immunosorbent assay (ELISA) was used to measure serum VEGF and PLGF levels during early ,mid and late pregnancy ,and those were compared among a‐bove groups .Results:Compared with normal pregnancy group ,there were significant reductions in serum levels of VEGF [ (187.54 ± 17.36) ng/L vs .(123.35 ± 16.46) ng/L] and PLGF [ (217.49 ± 26.32) ng/L vs .(128.32 ± 21.23) ng/L] in HDCP group during mid period (24th week) , P<0. 01 all ,and serum VEGF and PLGF levels more significantly reduced during late pregnancy (34th week) (P<0.01);in HDCP group ,compared with gestational hypertension group and severe preeclampsia group ,PLGF level [(127.46 ± 16.55) ng/L ,(158.40 ± 26.33) ng/L vs .(99.02 ± 27.26) ng/L] significantly reduced in mild preeclampsia group on 24th week;the serum VEGF and PLGF levels more significantly reduced on 34th week , P<0. 05 or <0. 01 . Conclusion:Reduced serum levels of VEGF and PLGF during mid pregnancy in HDCP patients are help to early diagnosing HDCP .

A 69-year-old male patient presented with a gradually enlarging mass in the left inner upper thigh for more than 2 months,and pigmented patches in the left medial leg for more than 20 years.Physical examination revealed a painless mass measuring 3 cm × 2 cm × 2 cm in size in the left inner upper thigh.Several pigmented patches were observed in the left medial leg,and the largest pigmented patch measured 2 cm× 2 cm in size with an irregular border and uneven pigmentation.The mass in the left inner upper thigh was resected and subjected to histopathological examination,which showed proliferative epithelioid neoplastic cells with mucous matrix,round and spindle cells of varying sizes separated by mucous matrix.The immunohistochemical study of tumor cells showed positive staining for vimentin,S100 and Melan-A,but negative staining for actin,desmin,CD56,epithelial membrane antigen,cytokeratin,leukocyte common antigen,CD99,chromogranin A and synaptophysin.Hematoxylin-eosin staining of pigmented patches on the left medial leg revealed squamous epithelium covering the surface of lesions with no superficial ulceration or atypia in epithelial cells,unevenly distributed melanophages,fibroplasia accompanied by collagen formation,obviously decreased skin appendages,infiltration of a few inflammatory cells in the dermis.AB-PAS staining was negative.The immunohistochemical study of pigmented patches showed positive staining for vimentin and Melan-A.The patient was pathologically diagnosed with metastatic myxoid melanoma with partial regression of the primary lesion.

Purpose To investigate the comparison of miR-106a in fresh and paraffin-embedded tissues and its expression in precan-cerous 1esions of gastric cancer. Methods Human gastric cancer tissues inc1uding 30 coup1es of fresh and 40 coup1es of paraffin-em-bedded samp1es were co11ected,quantitative rea1-time PCR was used to detect the expression of miR-106a in these two samp1es. Anoth-er paraffin-embedded samp1es inc1uding 20 cases of precancerous 1esions and 40 cases of gastric adenocarcinoma were a1so co11ected,in situ hybridization was used to assess the expression of miR-106a in these stages. Results The re1ative expression of miR-106a in fresh tissues was 3. 25 ± 1. 99,in paraffin-embedded tissues was 3. 18 ± 2. 14,indicating that the expression of miR-106a in these two sam-p1es has no statistica1 difference(P>0. 05),corre1ation ana1ysis showed that there was a significant corre1ation of miR-106a expression in these two samp1es(rs =0. 998,P<0. 001). The expression of miR-106a was detected in the stage of precancerous 1esions with the positive rate was 70%. The positive signa1s were 1ocated in dysp1astic epithe1ia1 ce11s and appeared as dark b1ue fine granu1es. The positive rate of miR-106a in gastric cancer tissues was 87. 5% and the frequency and extent increased and enhanced compared with pre-cancerous 1esions. Conclusion miR-106a has significant corre1ation in gastric cancer fresh tissues and paraffin-embedded tissues. Detection of miR-106a with paraffin tissues and its ear1y changes in prema1ignant 1esions can provide va1uab1e information for the ear1y diagnosis of gastric cancer.

Objective To describe the clinicopathologic features of two cases of mucoepidermoid carcinoma of the skin.Methods Two cases of mucoepidermoid carcinoma of the skin were analyzed histopathologically using hematoxylin and eosin (HE) staining,alcian blue-periodic acid Schiff (AB-PAS) staining and immunohistochemical staining.Relavant literature was reviewed.Results Histopathological examination showed that the tumor was subcutaneously located in both cases,with epidermoid cells and intermediate cells arranged in sheets or nests,as well as different sizes of glandular structures lined by mucinous columnar epithelium in some areas.Both tumors had a relatively clear boundary with peripheral invasive growth and no obvious capsules.Immunohistochemical examination showed positive staining for carcinoembryonic antigen (CEA),high and low molecular weight cytokeratin (CK(H) and CK(L)).The cytoplasm of mucous cells was stained blue with,and mucus was visualized after,AB-PAS staining.Conclusions Primary mucoepidermoid carcinoma of the skin is a kind of malignant tumor arising from skin appendages,whose diagnosis depends on histological and immunohistochemical examination.

Objective To develop an immunohistochemical assay for the diagnosis of cutaneous malignant melanoma (CMM) micrometastasis via blood and lymphatic vessels,and to evaluate its clinical significance.Methods Fifty-three patients (32 males and 21 females) histopathologically diagnosed as CMM were enrolled in this study.The patients were aged (61.2 ± 8.4) years (range,52-72 years).Tissue specimens were obtained from the central area of tumor in each case,and also from removed lymph nodes in some cases.The average duration of follow-up was (65.00 ± 5.68) months.During the follow-up,17 patients died of the recurrence or metastasis of CMM,and 6 patients were lost to follow-up.The expressions of D2-40,S100 and CD34 antigens in 53 tissue specimens were examined by immunohistochemical staining with three individual monoclonal antibodies,or by an immunohistochemical method using 2 two-antibody cocktails (D2-40/S 100 and CD34/S100) and double-color chromogens in single tissue sections.Results Of the 53 patients,30.19％ (16/53) were positive for hematoxylin-eosin (HE) staining combined with immunohistochemical staining with individual monoclonal antibodies,and 49.06％ (26/53) for the immunohistochemical method using two-antibody cocktails and double-color chromogens.Statistical differences were found in the positive rate between the two methods (x2 =3.94,P＜ 0.05).Compared with patients without blood/lymph vessel tumor emboli,those with blood/lymph vessel tumor emboli showed higher lymph node metastasis rate (80.77％ (21/26) vs.37.04％ (10/27),x2 =10.43,P ＜ 0.001),but lower five-year survival rate (42.31％ (11/26) vs.70.37％ (19/27),x2 =4.25,P ＜ 0.05).Conclusions The immunohistochemical method with two-antibody cocktails is superior to HE staining combined with immunohistochemical staining with individual monoclonal antibodies in the detection of blood/lymph vessel tumor emboli.And blood/lymph vessel tumor emboli may be an important prognostic factor in patients with CMM.

Objective To analyze the effect of clomipramine combine sleep deprivation on depression rat behavior and hippocampal CA3 neuron apoptosis.Methods Chronic unpredictable mild stimulation was used to establish 40 depression model rats.They were randomly divided into depression group,clomipramine group (5 mg/kg orally),sleep deprivation group (36 h continuous full sleep deprivation),chlorpromazine combination of sleep deprivation group,and then selected 10 rats as control group.Open-field test and water maze test were used to test rats' behavioral change after 28 days intervention.RT-PCR,Western blot were used to detect the GDNF and Bcl-2 expression of hippocampal CA3 neurons.TUNEL assay was performed to test hippocampal CA3 neuron apoptosis ratios in each group.Results Open-field test level score,vertical score and latency of combination group were (75.6±7.3),(26.4±4.3),(1.1±0.2),and showed no significant difference with those of control group((79.4 ±6.8),(28.6±5.7),(1.0±0.4)) after 28 d intervention (P＞0.05),while the level score,vertical score were significantly higher than those in other three groups(P＜0.05),and the latency period was significantly lower than that in other three groups (P＜0.05).The combined group showed a trend of gradually shortened in water maze escape latency.GDNF,Bcl-2 expression of control group and combined group was significantly higher than those in other three groups (P＜0.05),while the difference between the two groups was not statistically significant (P＞0.05).Sleep deprivation group' apoptotic rate was the highest (55±6)％,significantly higher than the control group and the combination group ((14±3) ％ and (19±4)％).Conclusion Clomipramine combined sleep deprivation can improve the ability of rat model of depression and suppression of acts of hippocampal CA3 neuron apoptosis,and its effect may be related to increased hippocampal CA3 GDNF,Bcl-2 level.