[The effect of platelet infusion on the repair of bone marrow hematopoietic niche damage in mice induced by (60)Co radiation and hematopoietic reconstruction after bone marrow transplantation].

Objective: To observe the effect of platelets on hematopoietic stem cell (HSCs) implantation in mice with radiation-induced bone marrow injury and bone marrow transplantation models. Methods: ①Male C57BL/6 mice were divided into a single irradiation group and a radiation infusion group after receiving (60)Co semimyeloablative irradiation for 18-10 weeks. The irradiation infusion group received 1×10(8) platelets expressing GFP fluorescent protein. ② The allogeneic bone marrow transplantation model was established. The experimental groups included the simple transplantation group (BMT) and the transplantation infusion group (BMT+PLT). The BMT group was infused through the tail vein only 5 × 10(6) bone marrow cells, the BMT+PLT group needs to be infused with bone marrow cells at the same time 1× 10(8) platelets. ③ Test indicators included peripheral blood cell and bone marrow cell counts, flow cytometry to detect the proportion of hematopoietic stem cell (HSC) and hematopoietic progenitor cells, bone marrow cell proliferation and apoptosis, and pathological observation of vascular niche damage and repair. Results: ①On the 3rd, 7th, 14(th), and 21st days after irradiation, the bone marrow cell count of the infusion group was higher than that in the single irradiation group (P<0.05), and the peripheral blood cell count was also higher. A statistically significant difference was found between the white blood cell count on the 21st day and the platelet count on the 7th day (P<0.05). In the observation cycle, the percentage of bone marrow cell proliferation in the infusion group was higher, while the percentage of apoptosis was lower. ② The results of bone tissue immunofluorescence after irradiation showed that the continuity of hematopoietic niche with red fluorescence was better in the irradiation infusion group. ③The chimerism percentage in the BMT+PLT group was always higher than that in the BMT group after transplantation.④ The BMT+PLT group had higher bone marrow cell count and percentage of bone marrow cell proliferation on the 7th and 28th day after transplantation than that in the BMT group, and the percentage of bone marrow cell apoptosis on the 14th day was lower than that in the BMT group (P<0.05). After the 14th day, the percentage of stem progenitor cells in the bone marrow cells of mice was higher than that in the BMT group (P<0.05). ⑤The immunohistochemical results of bone marrow tissue showed that the continuity of vascular endothelium in the BMT+PLT group was better than that in the BMT group. Conclusion: Platelet transfusion can alleviate the injury of vascular niche, promotes HSC homing, and is beneficial to hematopoietic reconstruction.

[Hepatic metastases after cervical cancer surgery: clinical analysis of 13 cases].

Objective: To investigate the clinical characteristics of patients with hepatic metastasis after cervical cancer operation, and analyze the risk factors and prognostic factors of hepatic metastasis. Methods: A total of 1 312 patients with stage Ⅰa2-Ⅱb2 cervical cancer received radical surgery from January 2013 to January 2016 in the First Affiliated Hospital of Zhengzhou University were collected, of which 13 cases (0.99%, 13/1 312) had hepatic metastasis after operation. A retrospective analysis was conducted on clinical features of patients with hepatic metastasis after cervical cancer operation. T-test, chi-square test, rank sum test and logistic regression were used to analyze the risk factors of hepatic metastasis after surgery of cervical cancer operation. Kaplan-Meier method was used for survival analysis. Log-rank test was used for screening of prognostic factors in patients with postoperative hepatic metastasis. Results: (1) Clinical features: there were 3 cases of simple hepatic metastasis without obvious clinical symptoms, 2 patients with perihepatic lymph node metastasis showed only low back pain, 8 patients with multiple extrahepatic metastases, and their clinical symptoms were related to the site of metastasis. Five cases out of 9 (5/9) with liver metastasis had abnormal tumor marker results. The abnormal kinds of tumor markers were mainly carcinoembryonic antigen (CEA), CA(125), CA(199), and CA(72-4). The interval time of hepatic metastasis after operation was 2-22 months. (2) Analysis of risk factors for hepatic metastasis: univariate analysis showed that lymph node metastasis, histological type, infiltration depth, and lymph-vascular space invasionwere associated with hepatic metastasis after cervical cancer surgery (P<0.05).Multivariate analysis showed that lymph node metastasis and small cell carcinoma were independent risk factors for postoperative hepatic metastasis (P<0.05). (3) Prognostic factors in patients with hepatic metastasis:among 13 patients with postoperative hepatic metastases from cervical cancer, 9 died during the follow-up period and 4 survived. The median total survival time after hepatic metastases was 7 months (range 3-32 months). Univariate analysis showed that multiple extrahepatic metastases and treatment after hepatic metastasis had significant effects on the prognosis of patients with hepatic metastasis after cervical cancer operation (P<0.05). Conclusions: The interval of hepatic metastasis after surgery for cervical cancer operation is within 2 years. Patients with lymph node metastasis and small cell carcinoma are more prone to postoperative hepatic metastasis. The prognosis of patients with extrahepatic multiple metastases is poor, and individualized treatment should be carried out after comprehensive analysis for patients with hepatic metastasis after cervical cancer operation.

[Comparative study of three-dimensional versus two-dimensional laparoscopic C1 radical hysterectomy for cervical cancer].

Objective: To compare the clinical effects and the subjective perception of surgeons with three-dimensional (3D) and two-dimensional (2D) laparoscopic C1 radical hysterectomy surgeries for cervical cancer. Methods: The retrospective cohort study was conducted. The clinicopathological data of 101 patients with cervical cancer who received C1 laparoscopic radical hysterectomy (C1-LRH) surgery from June 2015 to August 2017 were collected. Of all patients, 42 cases undergoing 3D laparoscopic surgery and 59 cases undergoing 2D laparoscopic surgery were respectively allocated into the C1-3DLRH group or C1-2DLRH group. The clinical effect and the subjective perception of surgeons were compared between the two groups. Results: (1) There was no significant difference between the C1-3DLRH group and C1-2DLRH group in terms of age, body mass index (BMI), International Federation of Gynecology and Obstetrics (FIGO) stage, pathologic type, etc. (all P>0.05). Compared with C1-2DLRH group, the operation time was significantly shortened [(192±54) vs (221±54) minutes, P<0.01], blood loss was significantly less [(102±88) vs (167±117) ml, P<0.01], and the success rate of inferior hypogastric plexus (IHP) bladder branch preservation was significantly increased [86% (36/42) vs 66% (39/59), P<0.05] in C1-3DLRH group. There were no significant difference in the number of lymph nodes, the incidence of operative complications, the infection rate, the time of catheterization and the length of hospitalization between the two groups (all P>0.05). The long-term bladder function was evaluated at the twelfth month after operation, 39 patients in the C1-3DLRH group and 53 patients in the C1-2DLRH group were completed the survey. The results showed that 13% (5/39) of the patients in the C1-3DLRH group had long-term bladder dysfunction, which was lower than that 21% (11/53) of the C1-2DLRH group, but there was no significant difference between the two groups (χ(2)=0.980, P=0.322). (2) A total of 251 laparoscopic surgeons questionnaires were eligible. The incidence of side effects in the first and second generation of 3D and 2D laparoscopic surgeons was 20.4% (10/49), 6.9% (6/87) and 3.5% (4/115), respectively. The incidence of side effects in the first generation of 3D laparoscopic surgeons was higher than that in the second generation of 3D (χ(2)=5.463, P=0.019) and 2D laparoscopic surgeons (χ(2)=12.475, P<0.01). There was no difference between the second generation of 3D and 2D laparoscopic surgeons (χ(2)=1.208, P=0.272). Conclusions: 3D laparoscopy is advantageous to the preservation of autonomic nerve in C1-LRH operation and may improve the quality of operation compared with 2D laparoscopy. The second generation of 3D laparoscopic device might overcome the side effects of the surgeons.

Effect of different time of ultrasound treatment on physicochemical, thermal, and antioxidant properties of chicken plasma protein.

The aim of present study was to investigate the effect of different times (5 min (UCPP-5), 10 min (UCPP-10), 20 min (UCPP-20), and 30 min (UCPP-30)) of ultrasound treatment on physicochemical, thermal, and antioxidant properties of chicken plasma protein (CPP). UCPP-20 had the highest fluorescence intensity and the lowest particle size. However, no major changes in the subunit compositions and the secondary structure of UCPPs were presented in SDS-PAGE and circular dichroism. The surface hydrophobicity and sulfhydryl content of UCPPs increased significantly (P < 0.05) as compared to those of CPP. With the increasing time of ultrasound treatment, there were more and deeper holes on the protein surfaces. Furthermore, protein modification by ultrasound could improve the thermal properties of UCPPs. Additionally, UCPPs showed a significant increase in antioxidant properties over CPP, especially UCPP-20. These observations indicated that ultrasound treatment was necessary for modification of CPP to meet the requirements for food processing.

MiR-381-3p inhibits proliferation, migration and invasion by targeting LRP6 in papillary thyroid carcinoma.

OBJECTIVE: MiR-381-3p plays an essential role in the progression of a variety of cancers, but its expression and role in papillary thyroid carcinoma (PTC) progression have not been investigated. The aim of this study was to investigate the expression of miR-381-3p and its function in PTC. PATIENTS AND METHODS: The expression levels of miR-381-3p and low-density lipoprotein receptor­related protein 6 (LRP6) mRNA in PTC tissues and cell lines were measured using RT-PCR. Cell proliferation, migration and invasion were assessed by cell viability assay and transwell assay. Luciferase assays and Western blotting were performed to demonstrate miR-381-3p target gene. RESULTS: We found that miR-381-3p was significantly down-regulated in PTC tissues and cell lines. In vitro assay indicated that up-regulation of miR-381-3p significantly suppressed PTC cell proliferation, migration and invasion. Moreover, luciferase reporter gene assay demonstrated that miR-381-3p could target LRP6 by binding to the 3' UTR. Western blot and Reverse transcription­quantitative polymerase chain reaction (RT­qPCR) showed that miR-381-3p overexpression suppressed the expression of LRP6 at both mRNA and proteins levels. In addition, functional experiment confirmed that LRP6 was involved in the suppressive effect of miR-381-3p-mediated PTC on cell proliferation, migration and invasion. CONCLUSIONS: Our findings suggested, for the first time, that miR-381-3p was lowly expressed in PTC tissues, and its up-regulation inhibited tumorigenesis of PTC by targeting LRP6.

[Prognostic analysis of stage â b2 andâ ¡a2 cervical squamous cancer without high risk factors treated with neo-adjuvant chemotherapy and radical hysterectomy].

Objective: To analyze the prognosis of Ⅰb2 and Ⅱa2 cervical squamous cancer without high risk factors treated with neo-adjuvant chemotherapy (NACT) and radical hysterectomy. Methods: This retrospective study included patients with International Federation of Gynecology and Obstetrics (FIGO) stage Ⅰ b2 and Ⅱ a2 cervical squamous cancer without high risk factors who underwent platinum-based NACT followed by radical surgery from January 2008 to January 2015. The responses of NACT were observed and compared in their effect on postoperative pathologic risk factors. Kaplan-Meier method and Cox regression analysis were performed to analyze survival status. Results: This study was recruited 282 patients with the average age of (44.4±6.7) years old. After NACT, 42 patients achieved complete response [CR, 14.9% (42/282) ], while 138 patients achieved partial response [PR, 48.9% (138/282) ] and 102 stable disease [SD, 36.2% (102/282) ]. The rate of pathologic diameter ≥4 cm, deep stromal invasion (DSI) positive and lymph-vascular space invasion (LVSI) positive rate decreased significantly in CR and PR group compared with SD group (P<0.05) . The number of postoperative risk factors in CR, PR and SD groups varied significantly (χ(2)=64.869, P=0.000) . Besides, the rate of multiple intermediate risk factors was respectively 0 vs 13.8% vs 45.1% (χ(2)=7.107, P=0.008) . The disease relapsed in 23 patients, and 12 died. On the whole series, 5-year overall survival rate was 91.7%, and 5-year recurrence-free survival rate was 88.9%. The rate of recurrence (P=0.002) and mortality (P=0.036) were higher in LVSI positive patients compared with LVSI negative. And the rate of recurrence was higher in patients with multiple intermediate risk factors, compared with no or one intermediate risk factors (P=0.002) . Univariate analysis revealed that LVSI positive and multiple intermediate risk factors were the factors predicted recurrence and mortality (P<0.05) , and no significance in age, stage, tumor grade, tumor diameter before or after NACT, response to NACT, or DSI positive factors (all P>0.05) . The multivariate analysis showed that, the factor of primary tumor diameter ≥6 cm (P=0.022) and multiple intermediate risk factors (P=0.001) were independent prognostic variables for recurrent-free survival. Besides, multiple intermediate risk factors was independent prognostic variable for overall survival (P=0.034) . After surgery, 107 patients did not receive adjuvant therapy, while 175 patients received adjuvant therapy with 26 radiotherapy, 46 chemotherapy and 103 concurrent radiochemotherapy. On survival analysis of postoperative adjuvant treatment, 5-year recurrence-free survival rate of radiotherapy group was significantly lower in patients with the factor of SD response to NACT (P=0.011) and multiple intermediate risk factors (P=0.008) , while overall no significance in overall survival rate (P>0.05) . Conclusions: NACT may be play beneficial role in relieving the status of intermediate risk factors for stage Ⅰ b2 and Ⅱ a2 cervical squamous cancer without high risk factors. The status of multiple intermediate risk factors is independent prognostic factors for recurrence and mortality. For patients with multiple intermediate risk factors, postoperative adjuvant chemotherapy or concurrent radiochemotherapy might be the better choice to prevent relapse.

[Application of liver three-dimensional visualization technologies in the treatment planning of hepatic malignant tumor].

Objective: To discuss the application of three dimentional(3D)visualization technologies in treatment plan of hepatic malignant tumor. Methods: The clinical data of 300 patients with liver malignant tumor who received treatment from January 2016 to January 2017 in the Third Department of Hepatic Surgery of Eastern Hepatobiliary Surgery Hospital was retrospectively analyzed in this study, including 221 male and 79 female patients aged from 7 to 76 years with median age of 54 years. The median height was 168 cm (115-183 cm), the median weight was 65 kg (20-105 kg) and the median tumor volume was 142 ml (23-2 493 ml). Three-dimensional visualization technology was used in all patients to reconstruct liver three-dimensional graphics. Also, two and three-dimensional methods were taken respectively to evaluate patients and develop treatment strategy. The change of treatment strategy caused by 3D evaluation, actual surgical plan, operation time, time of hepatic vascular occlusion, intraoperative blood loss, volumes of blood transfusion and postoperative complications was observed. Results: After three-dimensional visualization technology was applied, 75(25%) of 300 patients' treatment strategies had been changed. The range of hepatectomy was extended in 25 patients. And 7 of them were due to hepatic venous variation, which resulted in increasing drainage area. In other 4 patients, liver resections were extended due to lack of perfusion of the liver parenchyma after the removal of portal vein. And hepatectomy was expanded in 14 patients in order to increase the surgical margin. The range of hepatectomy was reduced in 8 patients, 4 of which were due to hepatic venous variation, such as hepatic vein of segment 4 or lower right posterior hepatic vein. The remaining 4 cases were because of insufficient residual liver volume.The surgical resection was performed in 278 cases, 257 of which received operation directly. Left hepatectomy was performed in 24 patients and right hepatectomy was performed in 33 patients. Left trisectionectomy was carried out in 12 patients and right trisectionectomy was carried out in 11 patients. Caudate lobectomy was applied in 10 patients. There were 18 cases of left lateral sectionectomy, 7 cases of right anterior sectionectomy, 25 cases of right posterior sectionectomy and 18 cases of mesohepatectomy. Single or multi segment resection was performed in 99 patients. The treatment strategy of thirty-six patients was converted to staged hepatectomy (ALPPS 11 cases and portal vein embolization 25 cases). The median operation time was 130 minutes (90-360 minutes) and the median inflow blood occlusion time was 20 minutes (0-75 minutes). Median blood loss volume was 200 ml (20-1 600 ml). Thirty-seven of 278 patients received transfusions, and the average red blood transfusion volume was (4.4±1.7)units (0-8 units). Median hepatic resection volume was 530 ml(30-2 600 ml). There were 117 cases of pleural effusion after operation, including 3 patients needing invasive therapy. Ascites occurred in 23 patients, 6 of whom needed invasive therapy. Biliary leakage was observed in 30 patients. Eight patients occurred hepatic cutting surface hemorrhage, 6 of whom received blood transfusion, and 4 of whom underwent laparotomy to stop bleeding. Three patients had pulmonary infection after surgery and 3 patients appeared biliary obstruction. Deep vein thrombosis took place in 2 patients and portal vein thrombosis was observed in 4 patients. No postoperative liver failure and death ever happened in our study group. Conclusion: Three-dimensional visualization technique can optimize the treatment strategy of patients with liver malignant tumor, improve surgical safety.

[Application of liver visualization technologies in hilar tumor resection at the second hepatic portal area].

OBJECTIVE: To discuss the application of liver visualization technology in complex liver tumor resection at the second hepatic portal area. METHODS: Clinical data of 80 cases who received surgery at the second hepatic portal area from August 2014 to September 2015 in the Third Department of Hepatic Surgery of Eastern Hepatobiliary Surgery Hospital were analyzed retrospectively. There were 58 male and 22 female patients aged from 21 to 70 years with median age of 52 years. Median maximum diameter of tumor was 7.6 cm (3.0 to 17.0 cm). Before surgery, liver dimensional graphics produced by liver visualization technology were taken on all patients to observe the relationship between intrahepatic vasculars and the liver tumor, and to calculate the intended resection range and the remaining liver volume in order to make a proper surgery plan. Suitable hepatic vascular occlusion was applied in the tumor resection. Intrahepatic vessel shape and variation, surgical operation, surgical operation time, manner and time of hepatic vascular occlusion, blood loss, liver resection volume, postoperative complications were observed. RESULTS: There were 23 patients who changed surgery plan after liver visualization technology.There were 44 cases with single main hepatic vein compressed by tumors, 32 cases with 2 main hepatic veins, 4 cases with 3 main hepatic veins compressed by tumors.And there were 58 cases with both hepatic vein and inferior vena cava compressed by tumor. Hepatic segments 6 and 7 was removed in 12 cases, 14 cases, hepatic segments 4, 5 and 8 were removed in 8 cases.Right hepatectomy was carried out in 9 patients and left hepatectomy was carried out in 8 patients. Right trisectionectomy was carried out in 3 patients and left trisectionectomy was applied in 5 patients.Local hepatectomy was performed in 12 patients. Nine patients received associating liver partition and portal vein ligation for staged hepatectomy. Four patients underwent total hepatic vascular exclusion, while 16 patients underwent selective hepatic vascular exclusion. The median surgical time was 132 minutes(80 to 240 minutes). Median blood loss volume was 580 ml(100-5 000 ml). Median volume of hepatic resection was 750 ml(30 to 2 000 ml). One patient needed secondary surgery to stop bleeding as a result of postoperative abdominal bleeding.Complication of postoperative bile leakage occurred in 14 cases.Five patients had pleural effusion requiring invasive therapy.Four patients had ascites requiring invasive therapy. Besides, 5 patients had incisive infection while 2 patients were found with pulmonary infection after surgery and two patients occurred biliary obstruction. There was no death case occurred a result of surgery. CONCLUSIONS: Using liver visualization technology to make surgical operation plan can improve surgical safety of the second hepatic portal area and optimize the operation plan. It can also reduce the risk of blood loss and postoperative complications such as liver failure.

[Expression and clinical significance of ELOVL6 gene in high-grade serous ovarian carcinoma].

OBJECTIVE: To investigate the expression of elongation of very long-chain fatty acids family member 6 (ELOVL6) in high-grade serous ovarian carcinoma (HSOC), and explore the correlation between its expression and clinical prognosis in these patients. METHODS: The expression of ELOVL6 at mRNA and protein levels were respectively detected by reverse transcription (RT)-PCR and immune histochemistry method in 12 cases with normal ovarian tissues and 172 cases with HSOC from primary tumor site, forty of which had paired peritoneal metastatic tissues. RESULTS: (1) The results tested by RT-PCR showed that ELOVL6 expression in normal ovarian tissue was 4.8±1.1, while 1.2±0.7 in primary tumors and 1.8±0.9 peritoneal metastatic sites in HSOC. Compared with normal ovarian tissue, the level of ELOVL6 mRNA was significantly lower in HSOC (P<0.05). There was no significant difference between primary and peritoneal metastatic sites in HSOC (P=0.610). It was shown that ELOVL6 protein localized in cytoplasm of ovarian cancer cell by immunostaining assay. (2) ELOVL6 expression was observed in all normal ovarian tissue, 70.2% of G1-G2 and 48.8% of G3 HSOC (P<0.05). ELOVL6 expression in drug-resistant group were significantly lower than that in non-resistant group (39.1% vs 65.0%, P<0.01). The median disease-free survival was 41 months in the ELOVL6-positive group and 39 months in ELOVL6-negative group (P>0.05). The total median survival was 52 months in ELOVL6-positive group and 44 months in ELOVL6-negative group (P>0.05). CONCLUSION: Low expression of ELOVL6 may correlate with the poor differentiation and drug resistance in HSOC.

[The experience of abundant amount of pectoralis major myocutaneous flap tissue in restoring parenchyma defect of head and neck].

Objective:Explore the skills to flap the defects of the head and neck with myocutaneous of pectoralis major which has rich amount of organization to minimize complications.Method:Various of neck defects will be repaired when the myocutaneous flap of pectoralis major from obese male or/and female has been prepared with improved methods. First of all, the muscle must be fixed. Then we have to suture the skin and the defect of skin or mucous membrane;the potential closed cavities around the myocutaneous flap must be drainaged with the negative pressure after repaired. Clinical data of 23 (24 sides) patients undergoing the repair in our hospital between January 2008 and December 2014 were collected and reviewed retrospectively.Result:One of 23 cases was injured the thoracoacromial artery in the preparation process of myocutaneous flap by the operator, then we operated on the other side. All of the pectoralis major myocutaneous flaps has been survived except 1 case which was necrosis in distal partial of pectoralis major muscle flap, but healed after changing dressing.Three cases with pharyngeal leakage because of the infection were also healed after changing dressing; skin and skin flap partial dehiscence happend in 1 cases, but also healed after dressing; 1 cases with subcutaneous hemorrhage was healed after cleaning up hematocele.Conclusion:The pectoralis major myocutaneous flap is still a good material for the repair of head and neck defects as long as the management skill in perioperative has been mastered properly, even though the one with rich amount of organization is bloated and the preparation and repair is difficult.

Protective Effects of Total Glucosides of Paeony on N-nitrosodiethylamine-induced Hepatocellular Carcinoma in Rats via Down-regulation of Regulatory B Cells.

Total glucoside of paeony (TGP), extracted from the root of Paeonia Lactiflora, has been known to show anti-inflammatory, anti-oxidative, hepato-protective and immuno-regulatory activities. The aim of this present study was to determine the anti-tumor effect of TGP against N-nitrosodiethylamine (DEN)-induced hepatocellular carcinoma (HCC) in rats, and to find the related mechanisms. Rat HCC model was established by intragastrically administrating with DEN (8 mg/kg). We found the number of tumor nodules and the index of liver and spleen were increased in the model group compared with the normal group, and was significantly decreased by TGP. Additionally, TGP obviously improved the hepatic pathological lesions induced by DEN, and decreased the elevated levels of serum alanine aminotransferase (ALT), glutamic oxalacetic transaminase (AST), alkaline phosphatase (ALP) and alpha fetoprotein (AFP) by DEN. Moreover, TGP decreased the level of B cell-activating factor (BAFF) and the proportion of IL-10-producing regulatory B cells (Bregs), and the decrease of BAFF by TGP is positively correlated to the decrease of IL-10-producing Bregs by TGP. These results suggest that TGP had a good therapeutic action on DEN-induced HCC rats, which might be due to its down-regulation of Bregs through reducing the level of BAFF.

Induction of long-term oscillations in the γ frequency band by nAChR activation in rat hippocampal CA3 area.

The hippocampal neuronal network oscillation at γ frequency band (γ oscillation) is generated by the precise interaction between interneurons and principle cells. γ oscillation is associated with attention, learning and memory and is impaired in the diseased conditions such as Alzheimer's disease (AD) and schizophrenia. Nicotinic acetylcholine receptor (nAChR) plays an important role in the regulation of hippocampal neurotransmission and network activity. It is not known whether nicotine modulates plasticity of network activity at γ oscillations in the hippocampus. In this study we investigated the effects of nicotine on the long-term changes of KA-induced γ oscillations. We found that hippocampal γ oscillations can be enhanced by a low concentration of nicotine (1µM), such an enhancement lasts for hours after washing out of nicotine, suggesting a form of synaptic plasticity, named as long-term oscillation at γ frequency band (LTOγ). Nicotine-induced LTOγ was mimicked by the selective α4ß2 but not by α7 nAChR agonist and was involved in N-methyl-d-aspartate (NMDA) receptor activation as well as depended on excitatory and inhibitory neurotransmission. Our results indicate that nAChR activation induced plasticity in γ oscillation, which may be beneficial for the improvement of cognitive deficiency in AD and schizophrenia.

A prospective randomized controlled trial to compare two methods of selective hepatic vascular exclusion in partial hepatectomy.

BACKGROUND AND AIM: Selective hepatic vascular exclusion (SHVE) has not been widely used because of difficulty in extrahepatic isolation of hepatic veins. This study aims to compare the results of SHVE using tourniquets or Satinsky clamps on major hepatic veins in partial hepatectomy for liver tumors involving the roots of hepatic veins. METHODS: Between June 2008 and March 2012, a randomized controlled trial was performed on patients undergoing liver resection to compare selective hepatic vascular exclusion using tourniquets or Satinsky clamps in partial hepatectomy. In the tourniquet group, the hepatic veins were completely isolated and occluded with tourniquets. In the Satinsky clamp group, the hepatic veins were dissected on the anterior and side walls only and they were clamped directly by Satinsky clamps. RESULTS: The time for dissecting hepatic veins was significantly shorter in the Satinsky clamp group (7.5 ± 6.6 min vs 21.3 ± 7.4 min) than the tourniquet group. In the tourniquet group, 5 hepatic veins could not be completely isolated and encircled. In 4 additional patients the hepatic vein was slightly torn during dissection. These 9 patients received successful occlusion using Satinsky clamps. In the Satinsky group, all occlusion of the hepatic vein was successful. There was a significant difference in the success rate in hepatic vein occlusion using the Satinsky and the tourniquet groups 60/60 vs 51/60, P = 0.0018. CONCLUSIONS: Both techniques of hepatic vein occlusion were safe and efficacious. As the use of Satinsky clamps is safer, easier and took less time, it is recommended.

First Report of Bipolaris papendorfii Causing Corn Leaf Spot in China.

Corn is the most important cereal crop in China, with over 34.94 million ha being cultivated in the country annually. However, fungal diseases are a major limiting factor in corn production. In August 2012, 20 ha of corn fields in Anhui Province were found to be heavily infected by fungi. The margin of the lesion was achlorotic, and the middle was yellowish white or off-white, which was similar to the corn Curvalaria leaf spot. The oval lesions were approximately 5 to 7 mm. Lesion tissue was removed from the border between symptomatic and healthy tissue. The surface was sterilized in 75% ethanol for 30 s and 0.1% HgCl2 for 1 min, after which the sample was washed three times in sterile distilled water. The isolate was purified and subcultured on potato dextrose agar (PDA) at 25 ± 2°C. The initial color of the colony was light brown, turning dark brown after being cultured for 7 days. The conidia were boat-shaped or inverted pear-shaped and were clearly bent to one side. The cells of both ends were slightly lighter and respectively ranged from 34.5 to 44.0 µm and 12.0 to 21.0 µm away from the base, with the second cell as the widest. The majority conidia had three or four false septates; isolates produced light brown to medium brown conidiophore, scattered or clustered, often branching, and exhibited bending. These morphological characteristics matched with the description of Bipolaris papendorfii reported by Zhang (3). A pathogenicity test was conducted with the two isolates on each of the 36 corns by spraying 2 ml spore suspension (106 conidia/ml). For the control treatment, 36 corns were inoculated with an equal volume of sterilized water. Inoculated plants were placed in a greenhouse from 29 to 33°C and 95% relative humidity. The typical 5 to 7 mm oval lesions were observed 7 days after inoculation, except on the control samples. Three replications of 36 corns were used for each treatment. The isolate was consistently 100% reisolated from the diseased tissue according to Koch's postulate. The isolate was found to be morphologically similar to B. papendorfii. Preliminary morphological identification of the fungus was confirmed by PCR assay using genomic DNA extracted from the mycelium of a 7-day-old culture on PDA at 25 ± 2°C. A 550-bp amplified region of the internal transcribed spacer (ITS) of rDNA was generated using ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3') universal primers (1). The ITS region (GenBank Accession No. KC592365) was then sequenced by Sangon Biotech (Shanghai, China), and displayed 99% nucleotide similarity with the rDNA-ITS of B. papendorfii (JQ753972.1) separately after BLASTn research in GenBank. Based on the symptoms, fungal morphology, ITS sequence, and pathogenicity testing, this fungus was identified as B. papendorfii. The pathogen could reportedly infect tobacco and cotton (2). To our knowledge, this is the first study to report that B. papendorfii can infect corn in China. This report will establish a foundation for the further study of B. papendorfii to address the disease effectively. Further studies will be conducted to determine the incidence of the disease and the severity of damage caused by B. papendorfii as well as determine a possible mode for controlling the spread of the disease. References: (1) Y. J. Cao et al. Chin. J. Trop. Crops 31:1098, 2010. (2) H. Deng et al. Mycosystema 21:327, 2002. (3) T. Y. Zhang. Chin. Fungi Chi. 30:21, 2010.

Simian virus 40 Vp1 DNA-binding domain is functionally separable from the overlapping nuclear localization signal and is required for effective virion formation and full viability.

A DNA-binding domain (DBD) was identified on simian virus 40 (SV40) major capsid protein Vp1, and the domain's function in the SV40 life cycle was examined. The DBD was mapped by assaying various recombinant Vp1 proteins for DNA binding in vitro. The carboxy-terminal 58-residue truncated Vp1DeltaC58 pentamer bound DNA with a K(d) of 1.8 x 10(-9) M in terms of the protein pentamer, while full-length Vp1 and carboxy-terminal-17-truncated Vp1DeltaC17 had comparable apparent K(d)s of 5.3 x 10(-9) to 7.3 x 10(-9) M in terms of the protein monomers. Previously identified on Vp1 was a nuclear localization signal (NLS) consisting of two N-terminal basic clusters, NLS1 (4-KRK-6) and NLS2 (15-KKPK-18). Vp1DeltaC58 pentamers harboring multiple-point mutations in NLS1 (NLSm1), NLS2 (NLSm2), or both basic clusters (NLSm1. 2) had progressively decreased DNA-binding activity, down to 0.7% of the Vp1DeltaC58 level for NLSm1. 2 Vp1. These data, along with those of N-terminally truncated proteins, placed the DBD in overlap with the bipartite NLS. The role of the Vp1 DBD during infection was investigated by taking advantage of NLS phenotypic complementation (N. Ishii, A. Nakanishi, M. Yamada, M. H. Macalalad, and H. Kasamatsu, J. Virol. 68:8209-8216, 1994), in which an NLS-defective Vp1 could localize to the nucleus in the presence of wild-type minor capsid proteins Vp2 and Vp3. This approach made it possible to dissect the role of the bifunctional Vp1 NLS-DBD in virion assembly in the nucleus. Mutants of the viable nonoverlapping SV40 (NO-SV40) DNA NLSm1, NLSm2, and NLSm1. 2 replicated normally following transfection into host cells and produced capsid proteins at normal levels. All mutant Vp1s were able to interact with Vp3 in vitro. The mutants NLSm1 and NLSm1. 2 were nonviable, and the mutant Vp1s unexpectedly failed to localize to the nucleus though Vp2 and Vp3 did, suggesting that the mutated NLS1 acted as a dominant signal for the cytoplasmic localization of Vp1. Mutant NLSm2, for which the mutant Vp1's nuclear localization defect was complemented by Vp2 and Vp3, displayed a 5,000-fold reduced viability. Analysis of NLSm2 DNA-transfected cell lysate revealed a 10-fold reduction in the level of DNase I-protected viral DNA, and yet virion-like particles were found among the DNase I-resistant material. Collective results support a role for Vp1 NLS2-DBD2 in the assembly of virion particles. The results also suggest that this determinant can function in the infection of new cells.

Molecular cloning of a novel isoform of diphosphoinositol polyphosphate phosphohydrolase: a potential target of lithium therapy.

The mechanisms underlying the therapeutic effects of lithium are largely unknown but may involve progressive adaptive alterations at the level of gene expression. Using differential display PCR, we identify a novel cDNA fragment, the expression of which was increased in the rat frontal cortex after 5 weeks of lithium administration. A full-length cDNA (2954-nt) was cloned by arrayed cDNA library screening, and sequencing of the clone revealed an open reading frame of 537-bp encoding a 179-residue protein. Amino acid sequence comparisons revealed that our clone is a member of the Nudix hydrolase family, with the highest percentage of homology (95%) being with a subtype of human diphosphoinositol polyphosphate phosphohydrolase, hDIPP2. Northern blot analysis revealed that chronic lithium treatment significantly increased rDIPP2 mRNA levels in frontal cortex, but not in hippocampus, midbrain, and cerebellum. The effect of lithium on rDIPP2 mRNA expression was not shared by two other anticonvulsant mood stabilizers, carbamazepine and valproate. Time-course studies showed that 1-week of lithium had no effect on rDIPP2 mRNA abundance in the frontal cortex. Our results suggest that DIPP2 may represent a biologically relevant target of lithium therapy, further supporting the notion that abnormalities in inositol phosphate metabolism may be significant in the pathophysiology and pharmacotherapy of bipolar disorder.

Role of simian virus 40 Vp1 cysteines in virion infectivity.

We have developed a new nonoverlapping infectious viral genome (NO-SV40) in order to facilitate structure-based analysis of the simian virus 40 (SV40) life cycle. We first tested the role of cysteine residues in the formation of infectious virions by individually mutating the seven cysteines in the major capsid protein, Vp1. All seven cysteine mutants-C9A, C49A, C87A, C104A, C207S, C254A, and C267L-retained viability. In the crystal structure of SV40, disulfide bridges are formed between certain Cys104 residues on neighboring pentamers. However, our results show that none of these disulfide bonds are required for virion infectivity in culture. We also introduced five different mutations into Cys254, the most strictly conserved cysteine across the polyomavirus family. We found that C254L, C254S, C254G, C254Q, and C254R mutants all showed greatly reduced (around 100,000-fold) plaque-forming ability. These mutants had no apparent defect in viral DNA replication. Mutant Vp1's, as well as wild-type Vp2/3, were mostly localized in the nucleus. Further analysis of the C254L mutant revealed that the mutant Vp1 was able to form pentamers in vitro. DNase I-resistant virion-like particles were present in NO-SV40-C254L-transfected cell lysate, but at about 1/18 the amount in wild-type-transfected lysate. An examination of the three-dimensional structure reveals that Cys254 is buried near the surface of Vp1, so that it cannot form disulfide bonds, and is not involved in intrapentamer interactions, consistent with the normal pentamer formation by the C254L mutant. It is, however, located at a critical junction between three pentamers, on a conserved loop (G2H) that packs against the dual interpentamer Ca(2+)-binding sites and the invading C-terminal helix of an adjacent pentamer. The substitution by the larger side chains is predicted to cause a localized shift in the G2H loop, which may disrupt Ca(2+) ion coordination and the packing of the invading helix, consistent with the defect in virion assembly. Our experimental system thus allows dissection of structure-function relationships during the distinct steps of the SV40 life cycle.

Associated disturbances in calcium homeostasis and G protein-mediated cAMP signaling in bipolar I disorder.

BACKGROUND: Evidence of extensive cross-talk between calcium (Ca(2+))- and cAMP-mediated signaling systems suggests that previously reported abnormalities in Ca(2+) homeostasis in bipolar I (BP-I) patients may be linked to disturbances in the function of G proteins that mediate cAMP signaling. METHODS: To test this hypothesis, the beta-adrenergic agonist, isoproterenol, and the G protein activator, sodium fluoride (NaF), were used to stimulate cAMP production in B lymphoblasts from healthy and BP-I subjects phenotyped on basal intracellular calcium concentration ([Ca(2+)](B)). cAMP was measured by radioimmunoassay and [Ca(2+)](B) by ratiometric fluorometry with fura-2. RESULTS: Isoproterenol- (10 microM) stimulated cAMP formation was lower in intact B lymphoblasts from BP-I patients with high [Ca(2+)](B) (>/= 2 SD above the mean concentration of healthy subjects) compared with patients having normal B lymphoblast [Ca(2+)](B) and with healthy subjects. Although basal and NaF-stimulated cAMP production was greater in B lymphoblast membranes from male BP-I patients with high versus normal [Ca(2+)](B), there were no differences in the percent stimulation. This suggests the differences in NaF response resulted from higher basal adenylyl cyclase activity. CONCLUSIONS: These findings suggest that trait-dependent disturbances in processes regulating beta-adrenergic receptor sensitivity and G protein-mediated cAMP signaling occur in conjunction with altered Ca(2+) homeostasis in those BP-I patients with high B lymphoblast [Ca(2+)](B).

Increased cyclic AMP-dependent protein kinase activity in postmortem brain from patients with bipolar affective disorder.

Previous observations of reduced [3H]cyclic AMP binding in postmortem brain regions from bipolar affective disorder subjects imply cyclic AMP-dependent protein kinase function may be altered in this illness. To test this hypothesis, basal and stimulated cyclic AMP-dependent protein kinase activity was determined in cytosolic and particulate fractions of postmortem brain from bipolar disorder patients and matched controls. Maximal enzyme activity was significantly higher (104%) in temporal cortex cytosolic fractions from bipolar disorder brain compared with matched controls. In temporal cortex particulate fractions and in the cytosolic and particulate fractions of other brain regions, smaller but statistically nonsignificant increments in maximal enzyme activity were detected. Basal cyclic AMP-dependent protein kinase activity was also significantly higher (40%) in temporal cortex cytosolic fractions of bipolar disorder brain compared with controls. Estimated EC50 values for cyclic AMP activation of this kinase were significantly lower (70 and 58%, respectively) in both cytosolic and particulate fractions of temporal cortex from bipolar disorder subjects compared with controls. These findings suggest that higher cyclic AMP-dependent protein kinase activity in bipolar disorder brain may be associated with a reduction of regulatory subunits of this enzyme, reflecting a possible adaptive response of this transducing enzyme to increased cyclic AMP signaling in this disorder.

Characterization of alpha(s)-immunoreactive ADP-ribosylated proteins in postmortem human brain.

ADP-ribosylation of the stimulatory G protein alpha subunit, alpha(s), has been demonstrated in a number of different mammalian tissues. However, little is known about the occurrence and role of this process in modifying alpha(s) levels/function in human brain. In the present study, endogenous and cholera toxin (CTX)-catalyzed [32P]ADP-ribosylated products were characterized in postmortem human temporal cortex by (1) immunoprecipitation with alpha(s) antisera (RM/1), (2) comparisons of immunoblots and autoradiograms of the [32P]ADP-ribosylated products, and (3) limited protease digestion. Of the three major endogenous [32P]ADP-ribosylated products (48, 45, and 39 kDa) in postmortem brain, the 48-kDa and 45-kDa bands were clearly identified as alpha(s-L) (long isoform) and alpha(s-S) (short isoform), respectively. RM/1 immunoprecipitated the 39-kDa [32P]ADP-ribosylated protein, and overlays of immunoblots and autoradiograms showed that this product corresponded to an alpha(s)-like-immunoreactive protein. Furthermore, limited protease digestion of the 39-kDa endogenous [32P]ADP-ribosylated band generated peptide fragments similar to both endogenous and CTX-catalyzed [32P]ADP-ribosylated alpha(s-S). Two major CTX-catalyzed [32P]ADP-ribosylated products were also identified as alpha(s-L) (52 kDa) and alpha(s-S) (45 kDa). These findings clearly demonstrate that alpha(s) is a substrate for endogenous and CTX-catalyzed [32P]ADP-ribosylation in postmortem human brain. Furthermore, a lower molecular weight alpha(s)-like immunoreactive protein is also expressed in human brain and is a substrate for endogenous but not CTX-catalyzed [32P]ADP-ribosylation.