Cytotoxic T lymphocytes elicited by dendritic cell-targeted delivery of human papillomavirus type-16 E6/E7 fusion gene exert lethal effects on CaSki cells.

Human papillomavirus (HPV) is the primary etiologic agent of cervical cancer. Consideration of safety and non human leukocyte antigen restriction, protein vaccine has become the most likely form of HPV therapeutic vaccine, although none have so far been reported as effective. Since tumor cells consistently express the two proteins E6 and E7, most therapeutic vaccines target one or both of them. In this study, we fabricated DC vaccines by transducing replication-defective recombinant adenoviruses expressing E6/E7 fusion gene of HPV-16, to investigate the lethal effects of specific cytotoxic T lymphocytes (CTL) against CaSki cells in vitro. Mouse immature dendritic cells (DC) were generated from bone marrow, and transfected with pAd-E6/E7 to prepare a DC vaccine and to induce specific CTL. The surface expression of CD40, CD68, MHC II and CD11c was assessed by flow cytometry (FCM), and the lethal effects of CTL against CaSki cells were determined by DAPI, FCM and CCK-8 methods. Immature mouse DC was successfully transfected by pAd-E6/E7 in vitro, and the transfecting efficiency was 40%-50%. A DC vaccine was successfully prepared and was used to induce specific CTL. Experimental results showed that the percentage of apoptosis and killing rate of CaSki cells were significantly increased by coculturing with the specific CTL (p <0.05). These results illustrated that a DC vaccine modified by HPV-16 E6/E7 gene can induce apoptosis of CaSki cells by inducing CTL, which may be used as a new strategy for biological treatment of cervical cancer.

Expression of p-Akt in ovarian serous carcinoma and its association with proliferation and apoptosis.

The aim of the present study was to determine the expression of p-Akt in ovarian serous carcinoma (OSC) and its association with proliferation and apoptosis. Paraffin-embedded tissues of patients aged between 35 and 64 years old without history of radiotherapy, chemotherapy and hormone therapy prior to surgery were collected. In total, samples included 12 ovarian serous cystadenomas (OSAs), 18 ovarian serous borderline tumors (OS-BTs) and 46 OSCs. Of the 46 OSC samples, 16 were well-differentiated, 20 were moderately differentiated and 10 were poorly differentiated, while 22 developed lymphatic metastases and 24 were metastasis-free. An additional 10 paraffin-embedded normal ovarian tissues (NOTs) were used as controls. Streptavidin-peroxidase immunohistochemistry assays were used to investigate the expression of p-Akt and cyclin D1 in the collected samples. Compared with NOT, p-Akt expression in the OS-BT and OSC groups, as well as cyclin D1 expression in the OSA and OSC groups, was significantly elevated (P<0.05). Compared with the OSA group, p-Akt expression in the OSC group was significantly elevated (P<0.01) and reversely associated with tumor differentiation (P<0.01), whereas cyclin D1 expression showed no correlation with tumor differentiation (P>0.05). The expression of p-Akt, caspase-3 and cyclin D1 was positively associated with lymphatic metastasis (r=0.334; P=0.023). The expression of p-Akt gradually increased with carcinoma development and was associated with differentiation and metastasis of OSC, revealing that the activation of the PI3K/Akt signaling pathway is involved in the development of OSC. Furthermore, the expression of cyclin D1 gradually increased in the NOT, OSA, OS-BT and OSC groups and was associated with tumor metastasis.

Silencing of Bmi-1 gene by RNA interference enhances sensitivity to doxorubicin in breast cancer cells.

The oncogene Bmi-1 is highly up-regulated in breast carcinoma and is found to be efficient in preventing apoptosis of the cancer cells. Doxorubicin is an important chemotherapeutic agent against breast carcinoma. However, the effective therapeutic response to doxorubicin is often associated with severe toxicity. The present study is targetted at developing a strategy to increase doxorubicin sensitivity to lower doses without compromising its efficacy. A stable cell line with a persistent silencing of Bmi-1 was established. MTT assay was performed to evaluate 50% inhibitory concentration (IC50) values of doxorubicin. Apoptosis was detected by FCM and the expression of related genes [phosphor-Akt (pAkt), totle-Akt (tAkt), Bcl-2 and Bax] was studied by Western blot. In vivo, the sensitivity of the tumor tissues against doxorubicin was evaluated by transplanted MCF-7 nude mice model and the apoptosis of tissue cells was detected by TUNEL assay. The expression of pAkt and Bcl-2 was down-regulated, whereas Bax was up-regulated in Bmi-1 silencing cells. The results obtained indicated that silencing of Bmi-1 can render MCF-7 cells more sensitive to doxorubicin which induced a significantly higher percentage of apoptosis cells in vitro and in vivo. All together these results clearly demonstrate that Bmi-1 siliencing combined treatment of doxorubicin might be a new strategy for biological treatment on breast cancer.

Spontaneous spinal epidural hematoma: analysis of 23 cases.

BACKGROUND: Spontaneous spinal epidural hematoma is a rare but disabling disease. To explore its characters and find out what factors influence the prognosis, we gave a retrospective analysis of 23 patients in our department in the past 8 years. METHODS: Spontaneous spinal epidural hematoma was diagnosed by taking MRIs of patients without surgical management or by histopathological examination. We registered patient's case history, laboratory examination, radiological image, treatment, pathological result, and prognosis after 3 months and gave them nonparameter analysis. RESULTS: Primary neurological status and progressive intervals have correlation with prognosis (P< .01), and the latter less than 12 hours predict worse prognosis (P= .032). Spinal edema in MRI predicts pessimistic prognosis (P= .013). Long hematoma predicts worse prognosis (P= .02). Preoperative interval, emphasized by other authors, has no statistical correlation with prognosis in this study (P= .832). Finally, patients with a single hematoma or hematoma mingled with other spinal disturbance have the same prognosis (P= .065). CONCLUSIONS: The primary neurological status, progressive interval, spinal edema, and size of hematoma will influence the prognosis of the patient with SSEH. The major treatment is surgical intervention, and it should be operated as soon as possible to avoid the aggravation of neurological status. Conservative treatment is not considered unless patient's neurological deficiency has relieved in the early period.

[Clinical features of spontaneous spinal epidural hematoma and influential factors of its prognosis].

OBJECTIVE: To explore the clinical features of spontaneous spinal epidural hematoma (SSEH) and to find out factors influencing its prognosis. METHODS: From September 1998 to October 2006, 23 patients with SSEH (10 males and 13 females) were treated. Their ages ranged from 10 to 69 years. The primary neurological status were classified as grade A in 7 patients, B in 2 patients, C in 4 patients, D in 9 patients and E in 1 patients according to ASIA grading system. The progressive intervals of their symptoms were divided as four period: less than 12 hours (12 patients), 12 to 24 hours (2 patients), 24 to 48 hours (3 patients) and more than 48 hours (6 patients). SSEH was diagnosed by MRI or by histopathological examination. The cases history, laboratory examination, radiological image, treatment, pathological result and prognosis were recorded and analyzed after 3 month. RESULTS: In 23 patients, there were 1 case of deterioration, 8 cases of no change, 9 cases of improvement and 5 cases of complete recovery. The gender had no correlation with prognosis (P>0.05). In the patients who had shorter progressive interval and more severe edema of spinal cord, the prognosis was worse (P<0.05). In the patients who had mild neurological deficit, the prognosis was good (P<0.01). In 17 patients undergoing surgery, the scores for prognosis was 1 point in 1 case, 2 points in 5 cases, 3 points in 6 cases and 4 points in 5 cases; the operation time had no correlation with prognosis (r = 0.056, P>0.05). In 6 patients undergoing conservative treatment, the scores for prognosis were 2 points and 3 points in 3 cases respectively. CONCLUSION: Prognosis of patient with SSEH is influenced by his primary neurological status, progressive interval, spinal edema and size of hematoma. The major treatment is surgical evacuation of hematoma as early as possible to break the aggravation of spinal function. Conservative treatment is not considered unless the neurological defects recovered in the early period.

Influencing factors for posttraumatic hydrocephalus in patients suffering from severe traumatic brain injuries.

OBJECTIVE: To detect the influencing factors for posttraumatic hydrocephalus in patients with severe traumatic brain injuries and provide theoretical reference for clinical treatment. METHODS: Retrospective study was made on 139 patients with severe traumatic brain injuries in our hospital. The patients were divided into two groups: hydrocephalus group and non-hydrocephalus group. Single factor analysis and multiple factor analysis were used to determine the related factors and hydrocephalus. Multiple factor analysis was conducted with logistic regression. RESULTS: Posttraumatic hydrocephalus was found in 19.42% of patients. Age(OR equal to 1.050, 95%CI: 1.012-1.090), decompressive craniectomy (OR equal to 4.312, 95%CI: 1.127-16.503), subarachnoid hemorrhage(OR equal to 43.421, 95%CI: 7.835-240.652) and continuous lumbar drainage of cerebrospinal fluid (OR equal to 0.045, 95%CI: 0.011-0.175) were screened out from nine factors as the influencing factors for posttraumatic hydrocephalus. CONCLUSIONS: Risk factors for PTH are as follows: age, decompressive craniectomy and subarachnoid hemorrhage (SAH). Continuous lumbar drainage of cerebrospinal fluid can greatly reduce posttraumatic hydrocephalus.

A comparative study on therapeutic method of traumatic epidural hematoma.

OBJECTIVE: To explore the therapeutic methods, surgical indications and clinical practice of minimally invasive surgery on traumatic epidural hematoma (EDH). METHODS: Retrospective study was made on 135 patients with traumatic EDH admitted into our hospital from June 2002 to August 2005. Sixty-five patients were treated with mini-invasive negative pressure drainage (treatment group), 70 patients with comparable condition used traditional craniotomy (control group). The mean time of operation, average days in hospital, expenditure and prognosis of two groups were recorded and analyzed. RESULTS: There was no significant difference in therapeutic efficacy between two groups. Patients in treatment group had a shorter hospital stay and less expenditure than those in control group. CONCLUSION: Mini-invasive negative pressure drainage is simple, effective, economical and applicable to some traumatic EDH patients.

Effect of hyperbaric oxygen on cytochrome C, Bcl-2 and Bax expression after experimental traumatic brain injury in rats.

OBJECTIVE: To explore the effects of hyperbaric oxygen (HBO) treatment on the neuronal apoptosis at an earlier stage and the expressions of Cytochrome C (Cyt C), Bcl-2 (B-cell lymphoma-2 family) and Bax (Bcl-2 associated X protein) in rat brain tissues after traumatic brain injury (TBI). METHODS: Forty adult rats were divided into two groups, i.e., Group A (the rats with untreated TBI) and Group B (rats with HBO treatment after TBI). Sections of brain tissues of these two groups were then detected at 3, 6, 12, 24, 72 hours after TBI by immunohistochemistry and electronmicroscope, respectively. RESULTS: HBO treatment could up-regulate the expression of Bcl-2 within 72 hours, reduce the release of Cyt C from mitochondria, attenuate the formation of dimeric Bax and alleviate the mitochondrial edema within 24 hours after TBI. CONCLUSIONS: HBO treatment can alleviate neuronal apoptosis after TBI by reducing the release of Cyt C and the dimers of Bax and up-regulating the expression of Bcl-2.

[Changes in P-selectin expression after brain injury in rats].

OBJECTIVE: To explore the relationship between expression thange of P-selectin after brain injury and secondary brain damage. METHODS: Sixty SD rats were randomized into 3 equal groups, namely the control group, mild injury group and severe injury group and animal models of brain injury were established in SD rats according to the method of Feeney. P-selectin expression in the brain tissues were determined at 6 h and l, 3, and 7 days following brain injury (n=5 for each time point). Imaging analysis was performed using computerized imaging technique. RESULTS: P-selectin expression and neutrophil infiltration in the brain tissues increased significantly 6 h after brain injury (P<0.05), reaching the peak level at postoperative 24 h and then gradually decreased. CONCLUSION: P-selectin expression and neutrophil infiltration increase significantly following brain injury, and the time course and distribution of P-selectin expression are consistent with the secondary damage of the brain, strongly suggesting the involvement of P-selectin upregulation in the secondary insult after brain injury.

[Modified hemispherectomy for intractable epilepsy in patients with infantile hemiplegia].

OBJECTIVE: To explore the effectiveness of modified hemispherectomy for intractable epilepsy in patients with infantile hemiplegia. METHODS: Eighteen cases of patients were treated with modified hemispherectomy and the effectiveness was studied and followed up. RESULTS: The seizures in all 18 cases of patients were controlled effectively and stopped completely in 16 cases of them, without nervous disfunction worsened. The patients' cerebral peduncles on healthy side were much thicker than those on sick side (t = 58.32, P < 0.001) and healthy peoples' (t = 14.63, P < 0.001) and the patients' cerebral peduncles on sick side were much thinner than those of healthy peoples' (t = 51.27, P < 0.001). CONCLUSION: The modified hemispherectomy can effectively control the seizures of patients with infantile hemiplegia without superficial cerebral hemosiderosis happened.

[Effects of extract of Ginkgo biloba on intercellular adhesion molecule-1 and mRNA expression in rats with cerebral injury].

OBJECTIVE: To investigate the effects of Extract of Ginkgo biloba on ICAM-1 and mRNA expression in rats with cerebral injury, and discuss its protective mechanism. METHODS: The acute closed brain injury model was set up in rats according Feeney's method. Seventy-two rats were randomly divided into four groups. The levels of ICAM-1 expression were observed using immunohistochemistry and computerized imaging technique and the expression of mRNA were studied using RT-PCR techniques. RESULTS: ICAM-1 positive cells located in the transitional zone between the necrotic core and normal cortex. ICAM-1 protein and mRNA expression began to show 6 hours after cerebral injury, peaked at 24 hours. As compared with the model group, ICAM-1 and mRNA expression was significantly reduced in the group EGb (P < 0.01). CONCLUSION: EGb can markedly reduce the injury of nerve cell in the transitional zone, alleviate rat cerebral injury, and decrease ICAM-1 and mRNA expression and neutrophils infiltration. The protection may be related with its inhibition on ICAM-1 and mRNA expression.