BACKGROUND: Neuroprotective role of hypothermia on cerebral ischemic-reperfusional impairment has been long acknowledged. Since general hypothermia is complicated and unfit for observing postoperative consciousness and neurological function, it is of important significance to explore novel methods of focal cerebral hypothermia.OBJECTIVE: To study the neuroprotective effect of lateral ventricle infusion of low-temperature fluid on ischemic neurons of middle cerebral artery (MAC) occlusion models established on New Zealand rabbits.DESIGN: A randomized case-control study based on experimental animal models.SETTING: Neurosurgical department and pathological department of a general military hospital of Chinese PLA.MATERIALS: This study was carried out at Neurosurgical Laboratory of Nanjing General Hospital, Nanjing Military Area Command of Chinese PLA. Altogether 18 healthy New Zealand male rabbits, weighing from 2. 8 to 3.2 kg, were selected 4 - 6 months after birth, and randomly divided into occlusion group, hypothermia group and control group.INTERVENTIONS: Cerebral focal ischemic-reperfusional model was established on the New Zealand rabbits through MCA occlusion for 2 hours followed by reperfusion for 24 hours.MAIN OUTCOME MEASURES: Scores for neurological function, water content in the left and right brain, pathological changes of nerve cells in the left MCA supplying region.pothermia group, significantly higher than that in occlusion group(7.58 ± 0.58 )( P ＜ 0.01 ), but no significant difference could be observed in contrast with brain was(81.64 ± 0.82)% and (79.26 ± 1.30)% in occlusion and hypothermia groups with significant difference between them( P ＜ 0.05), and it was significantly different between the left side [ (81.64 ± 0. 82 )% ] and opyknosis and deep staining could be observed in nerve ganglion cells in occlusion group under optical microscope, but no obvious pathological changes were observed in MCA supplying brain regions in hypothermia group.CONCLUSION: Permanent infusion of low-temperature fluid into the lateral ventricle plays an important neuroprotective role by attenuating cerebral ischemic-reperfusional impairment and improving post-ischemic neurological functions.

Objective To evaluate the clinical efficacy and adverse effect of Traditional Chinese Medicine and acupuncture in treatment of ankylosing spondylitis. Methods 46 patients of ankylosing spondylitis with informed consent were randomly divided into observation group and control group by number table,each of 23 cases.The control group received traditional Chinese medicine,the observation group was treated with acupuncture and traditional Chinese medicine,the two groups were treated for 1 month,the clinical effect were observed. Results In the observation group,excellent in 18 cases(78.3%),effective in 3 cases(13.0%),ineffective in 2 cases(8.7%),the total effective rate was 91.3%,it was significantly higher than that of control group the 11 cases(47.8%),8(34.8%),4 cases (17.4%)and 82.6%,the difference between the groups was statistically significant(x2=4.572,P＜0.05).The patients of observation group had a rash in 1 case,1 case of pain;in the control group,1 case of rash,1 case of itching;all of the symptoms were relatively minor,did not affect the treatment.There were no significant adverse reactions in the patients of two groups. Conclusion Traditional Chinese medicine and acupuncture in treatment of ankylosing spondylitis had satisfactory effects and no significant adverse reactions,it was worthy to be recommended in the clinical application.

Objectives:To explore the protective effects of continuous trickle of low temperature liquids through the lateral ventricle on neurons after rabbit local cerebral ischemia reperfusion. Methods: The middle cerebral artery (MCA) of New Zealand rabbit was clipped with micro aneurysm clip for 2 hours and reperfused for 24 hours. Immediately after clipping the MCA, we trickled the left lateral ventricle continuously with low temperature liquids to decrease the brain temperature to mild hypothermia(32-35℃) and maintained for 2 hours. After reperfusion for 24 hours, we assessed the animal's neural deficits, observe the pathology of the ischemic brain tissues dyed by HE and determined the dry wet ratio for brain edema in the sham operated group, the control group and the mild hypothermia group respectively.Results:①The grades of neural deficits in mild hypothermia group were obviously lower than that in the clipping group( P 0.05 );②The dry wet ratios were obviously different in the mild hypothermia group and clipping group;③ Pyknosis and dense dying by HE were observed in the neural nuclei of ischemic cortex tissues of the clipping group, but no obvious changes were observed in the mild hypothermia group. Conclusions:Trickling ventricle with low temperature liquids could alleviate cerebral ischemia reperfusion injury, ameliorated neural deficit after ischemia reperfusion and were protective on the brain.

Objective: To explore the effects of continuous trickle of low temperature liquids through the lateral ventricle on neural cell apoptosis after rabbit local cerebral ischemia/reperfusion. Methods:The middle cerebral artery (MCA) of New Zealand rabbit was clipped by micro aneurysm clip for 2 hours and reperfused for 24 hours. Immediately after clipping the MCA, we trickled the left lateral ventricle continuously with low temperature liquids(22℃) to decrease the brain temperature to mild hypothermia (33℃-35℃)and maintained for 2 hours. After reperfusion for 24 hours , we assessed TUNEL method to determine the apoptotic cell rate in the sham-operated group, the control group and the mild hypothermia group respectively. Results:The apoptotic cell rate of the cortex tissues accommodated by MCA in the mild hypothermia group was obviously lower than that in the control group(P0.05). Conclusion:Trickling ventricle with low temperature liquids could decrease the apoptotic cell rate and alleviate cerebral ischemia/reperfusion injury.

Objective To explore the characteristics and treatment of pain after inflammatory spinal demyelination. Methods 271 patientssuffered from inflammatory spinal demyelination with pain were analyzed retrospectively. Results Acute radicular pain and Lhermitte'ssign were common in the acute pain syndromes. Individual therapy showed a benefit of decreased pain. Conclusion Pain is a commonclinical symptom of inflammatory spinal demyelination. Individualized therapeutic decisions could relieve symptom and improve outcome.

Objectives:To study the clinicopathological features of extraovarian peritoneal serous papillary carcinoma.Methods:The clinical pictures and pathological characteristics of 6 cases extraovarian peritoneal serous papillary carcinoma admitted to our hospital from 1992 to 1999 were studies retrospectively.H & E staining,histochemical staining and immunohistochemical staining were used.Results:The ages of the 6 patients were between 32～62(43.7) years old.The bilateral ovaries and fallopian tubes were almost normal.Tumors were examined histomorphologically by microscope and electron microscope,periodic acid-Schiff (PAS) stain was used in the histochemical study and carcinoembryonic antigen (CEA),CA125,p53,AE1 and AE3 monoclonal antibody assays were detected immunohistochemically.The results revealed no difference between extraovarian peritoneal serous papillary carcinoma and ovarian serous papillary carcinoma.Conclusions:Extraovarian peritoneal serous papillary carcinoma is considered arising from the secondary Müllerian system.Histologically,extraovarian peritoneal serous papillary carcinoma was identical to that of the similar epithelial carcinomas arising from the ovaries.The diagnosis was made only when the ovaries were not involved and without any evidence of similar epithelial carcinoma obtained in the ovaries.Histochemical staining and immunohistochemical staining are helpful as accessory criteria for the differential diagnosis between primary carcinoma of the peritoneum and malignant mesothelioma.The prognosis is poor.

Objective　To study the histopathologic features and pathogenesis of mucosa-associated lymphoid tissue lymphoma (MALT-oma) of salivary glands. Methods　Clinical data, paraffin-embedded sections, immunohistochemical slides (SP method) and electron microscopic features of surgical specimens of 32 cases of salivary gland MALT-oma were studied. Results　The patients were 27 males and 5 females, with a mean age of 54.76 years. The lesions were located in the parotid area in 17 cases, and in the submandibular gland in the remaining 15 cases. Much of the MALT-oma was replaced by infiltration of a great amount of centrocyte-like cells (CCL) as background and occasional large cells (centroblast- or immunoblast-like). In MALT-omas “lymphoepithelial lesions" were present. Immunohistochemically, CD20 expression was found to be positive and CD45RO expression was negative in all MALT-omas. Conclusion　Most of the MALT-omas are low grade malignant tumors and have a “homing back" phenomenon. The cases were managed by surgery and chemotherapy. In a few MALT- omas which turned into high grade malignant tumors, the prognosis was poor. Acquired MALT may develop as a reaction to autoimmune disease and infection. Hyper-immune reaction and MALT hyperplasia under stimulation may result in myoepithelial sialadenitis and lead to MALT-oma of the salivary gland.

Objective　To analyse the clinical manifestations, pathoogical and immunohistochemica l features of 9 cases of primary cutaneous CD30-positive anaplastic large cell ly mphoma (ALCL) in order to improve the criteria for its early and differential diagnosis. Methods　 Histopathological method was used to study the morphological characteristics. Immunohistochemical stainings (SP or ABC method) for leukocyte common antigen (LCA), CD20, CD30, CD45RO, CD68, epithelium membran e antigen (EMA), cytokeratin (CK) and HMB45 were applied to the routine para ffin sections of all the 9 cases. Results　 The 9 cases of primary cutaneous CD30-pos itive ALCL consisting of 6 males and 3 females, aged 31 to 84 years (mean 58.2 years). All patients presented with subcutaneous masses or papular eruptions on lower trunk and extremities. Histopatholoically, the lesions were composed of nu merous large round, oval or pleomorphic cells with abundant cytoplasm, amphophil ic or basophilic, and large nucleus with distinct nucleolus. Mitosis was often o bserved. Multinucleated giant cells and Reed-Sternberg cells may be seen. The n e oplastic cells displayed positive antigen markers for CD30 in 9 cases, 6 positiv e for CD45RO and 3 cases negative for both CD45RO and CD20. Two patients died of metastasis, 2 patients experienced recurrence and 5 patients still well during follow-up. Conclusions　 Primary cutaneous CD30-positive ALCL is a morphologically distinctive neoplasm with a comparatively favorable prognosis. This tumor can be differentiated from other malignant tumors on the basis of histopathologic feat ures and positive CD30 which is of significance.

Objective: To evaluate the anti-tumor activity of mouse multi-subtype heat shock protein/peptide (mHSP/P) vaccine in combination with a programmed death ligand 1 (PD-L1) inhibitor in mouse sarcoma. Methods: Immunohistochemical staining and en-zyme-linked immunosorbent assay (Elisa) was used to quantitatively identify the expression of heat shock proteins (HSP70, HSP90, Grp94) in the sarcoma cell line MCA207. From the protein suspension prepared, mHSP/P and Grp94/peptide (Grp94/P) sarcoma vac-cines were isolated using chromatography and were identified by Western blot (WB). Flow cytometry was used to determine their cy-totoxic effects. The levels of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) produced upon mHSP/P and Grp94/P stimulation were measured by Elisa. The effect of sarcoma vaccines on the growth and survival of sarcoma was evaluated in mice. The expression of PD-L1 on the surface of MCA207 sarcoma cells was evaluated by immunofluorescent staining. The effect of IFN-γ treatment on the expression of PD-L1 was determined by WB. Animal experiments explored the effects of PD-L1 inhibitor in combination with mHSP/P treatment on tumors. Results: Tumor tissue carries a variety of HSP subtypes (HSP70, HSP90, Grp94). We successfully isolated sarco-ma tissue-derived mHSP/P and Grp94/P tumor vaccines, which were identified by WB; flow cytometry analysis demonstrated their cy-totoxicity. The levels of IFN-γ and TNF-α cytokines upon mHSP/P stimulation were significantly higher than that observed upon Grp94/P stimulation (P<0.05). The expression of PD-L1 on the surface of sarcoma cells increased with IFN-γ treatment. Animal experiments demonstrated that PD-L1 inhibitor in combination with mHSP/P significantly increased the immune response against tumor (P<0.05). Conclusions: Tumor-derived mHSP/P and Grp94/P can be used as tumor vaccines in animal models. The mHSP/P can elicit a stronger anti-tumor immune response than Grp94/P. IFN-γ stimulates the expression of PD-L1 in sarcoma cells, which results in immune eva-sion. The PD-L1 inhibitor in combination with mHSP/P increased the anti-tumor effect in the tumor microenvironment.

In this study, we aimed to construct a non-replication mRNA platform and explore the side effects of electroporation-mediated delivery of mRNA on the mice as well as the expression features of the mRNA. With luciferase gene as a marker, in vitro transcription with T7 RNA polymerase was carried out for the synthesis of luciferase-expressed mRNA, followed by enzymatic capping and tailing. The mRNA was delivered in vivo by electroporation via an in vivo gene delivery system, and the expression intensity and duration of luciferase in mice were observed via an in vivo imaging system. The results demonstrated that the mRNA transcripts were successfully expressed both in vitro and in vivo. The electroporation-mediated delivery of mRNA had no obvious side effects on the mice. Luciferase was expressed successfully in all the mRNA-transduced mice, while the expression intensity and duration varied among individuals. Overall, the expression level peaked on the first day after electroporation and rapidly declined on the fourth day. This study is of great importance for the construction of non-replication mRNAs and their application in vaccine or antitumor drug development.
Animals ; Electroporation/methods* ; Gene Transfer Techniques ; Luciferases/metabolism* ; Mice ; RNA, Messenger/genetics*