Objective: To investigate the effects of novel targeted non-viral vector in gene therapy of human glioma. Methods: The EGF-R targeting gene delivery system GE7 was constructed. Human Glioma cell line U251 was transfected in vitro with ?－gal as reportor gene and p21 as therapeutic gene using this gene delivery system. By means of the assay of ?-galactosidase staining, Western blotting, in situ end labeling apoptosis cells and DNA ladder, the transferring of exogenous genes and the apoptosis of the tumor cells were examined.Results: It was showed that gene transfer efficiency is over 80%. When transfected with p21 gene, the growth of cells was inhibited significantly, and the apoptosis was detected in the transfected cell by the methods of in situ end labeling and DNA ladder. Conclusion: The GE7 gene delivery system has the ability to transfer exogenous gene to tumor cells and the expression of the therapeutic gene can inhibit the growth of the cells.

Objective To investigate gene transfer efficiency of a novel target non-viral vector GE7 and effects of herpes simplex virus thymidine kinase (HSV 1-tk)/ganciclovir(GCV) mediated by it in vitro. Methods The epidermal growth factor receptor (EGF-R) target gene delivery system GE7 was constructed.Human ovarian cancer cell line CAOV3 was transfected in vitro with ?-galactosidase(?-gal) as reporter gene and HSV 1-tk gene as therapeutic gene using this gene delivery system.By means of the assay of X-gal staining, Northern blotting, cell growth-inhibiting curve and so on,the transferring efficiency of exogenous genes and killing effects are observed. Results It showed that gene transfer efficiency is over 80%.When 10 mg/L GCV was put into ovarian cells transfected with HSV 1-tk gene, 95% of cells were killed, and the apoptosis ratio reached up to 30. Conclusions The GE7 gene delivery system is an effective and safe delivery system.GE7/ HSV 1-tk /GCV therapeutic gene system is appraising for ovarian cancer.

Objective To sum up our experience on the diagnosis and treatment of rare hepatic tumors. Methods The data of 25 patients with rare liver tumors admitted in our hospital from May 2005 to January 2010 were analyzed retrospectively. Results The final pathologic diagnosis of focal nodular hyperplasia was made in 6 cases, and the diagnosis of vascular leiomyoma, hilar neurilemoma, intrahepatic aneurysm, biliary cystadenoma, hepatic hamartoma, biliary villous adenoma, and hepatic diffuse large B-cell lymphoma was established in one each case, respectively. The diagnosis of angiomyolipoma in 2patients, primary liver gastroimestinal stromal tumor in 2 patients, hepatoblastoma in 5 patients and liver undifferentiated sarcoma in 3 patients was established. Preoperative ultrasonography, CT and MRI were performed in 24, 22 and 6 patients respectively. Preoperative tentative diagnosis was finally confirmed by pathology in only 3 (16.7%) cases, all by CT report. Preoperative diagnosis was consistent with postoperative pathology in 5 patients (20%); All patients underwent liver resection including hemihepatectomy in 7 patients, hepatic lobectomy in 7 patients, segmentectomy in 9 patients and tumor enucleation in 2 patients; There was no recurrence after resection of benign, low malignant tumors and hepatic diffuse large B-cell lymphoma; Postoperative follow-up was made for all the 5 cases of malignant tumours, and there was recurrence in 3 cases. These 3 eases underwent second resection and there were no recurrences after reoperation. The two recurrent patients died with a mean survival of 4 months.Conclusions The preoperative correct imaging diagnostic rate for rare hepatic tumors is low. Surgery is the most effective therapy and reoperation should always be attempted for tumor recurrence in order to prolong survival.

Objective To explore further the abilities of cerebellar lesions in skills of time estimation, and to test the hypothesis that cerebellum is involved in the special network of time perception. Methods Time reproduction was required for 3 time intervals of 600-milliseconds, 3 and 5 seconds with visual discrimination for control. Participants reproduce those time intervals after 1-second or 5-seconds of delay. Twenty-six patients with cerebellar lesions were compared to 26 healthy controls, matched for age, handedness, education. Results 600-milliseconds was overestimated by both cerebellar lesions and healthy controls, however, both 3-seconds and 5-seconds were underestimated. Patients with cerebellar lesions were significantly impaired on 600-milliseconds reproduction task ( delaying 1 s or 5 s, 1.37 ± 0.24, 1.26 ± 0.29 respectively, Z=-5.347, -4.230, both P<0.01). No group differences were found for the 3-seconds and 5-seconds time reproduction (delaying 1 s or 5 s, Z=-1.200,-0.092,-1.519, -0.723, all P>0.05). Conclusion The findings suggest that patients with cerebellar lesions perform poorly during measurement of the shorter interval, but show no impairment of longer intervals perception, supporting the hypotheses that cerebellum is specifically involved in the perception of sub-second intervals.

OBJECTIVESTo construct a hepatoma directed gene delivery system which could transfer preS2 antisense RNA to liver cancer cells specifically, and to explore a new therapeutic strategy for hepatocellular carcinoma by blocking hepatitis B virus (HBV) with antisense RNA targeting hepatocellular carcinoma.

METHODSGE7 and HA20 were synthesized and mixed with pEBAF-as-preS2, a hepatocarcinoma specific HBV antisense expression vector, to construct a novel HBV antisense RNA delivery system named AFP-enhancing 4-element complex. Nude mice bearing hepatocelluar carcinoma cells HepG2.2.15 were injected with AFP-enhancing 4-element complex via a tail vein. Total RNA from tissues was extracted, and reversal transcription-ploymerase chain reaction (RT-PCR) was used to detect the expression of preS2. Different doses of AFP-enhancing 4-element complex was injected into nude mice at different time points, and tumor diameter was measured.

RESULTSAFP-enhancing 4-element complex was constructed successfully. RT-PCR showed preS2 antisense RNA delivered by AFP-enhancing 4-element complex only expressed in liver tumor HepG2.2.15 cells of the mice. After the treatment of AFP-enhancing 4-element complex with dose of 0.2 micro g per mouse (once a week for 4 weeks), the mean tumor diameter of nude mice was significantly shorter than that of the control groups (0.995 +/- 0.35 cm vs 2.125 +/- 0.25 cm, P < 0.01).

CONCLUSIONSAn HBV antisense RNA gene delivery system targeting hepatocellular carcinoma, AFP-enhancing 4-element complex, was constructed successfully. PreS2 antisense RNA expressed specifically in hepatocelluar carcinoma cells significantly inhibits tumor growth of mice bearing hepatocarcinoma HepG2.2.15 and may have therapeutic potential in HBV related hepatocarcinoma.

Animals ; Drug Delivery Systems ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; Hepatitis B Surface Antigens ; therapeutic use ; Hepatitis B virus ; genetics ; Liver Neoplasms, Experimental ; pathology ; therapy ; Male ; Mice ; Mice, Nude ; Protein Precursors ; therapeutic use ; RNA, Antisense ; therapeutic use

Objective:To investigate the curative efficacy and application value of convalescent plasma (CP)in severe and critical coronavirus disease 2019 (COVID-19) caused by Delta variant.Methods:The treatment process and results of CP therapy for a patient with critical COVID-19 caused by Delta variant were reported. The clinical application value of CP for COVID-19 caused by Delta variant was analyzed along with the literature review.Results:Our case was a 50-year-old male, who was imported from abroad and had not been vaccinated against COVID-19. The novel coronavirus nucleic acid test was negative before entry. On the second day after entry, fever occurred, novel coronavirus nucleic acid test was positive. Chest CT images showed bilateral multiple mottling and ground-glass opacity with symptoms of nausea, headache, loss of appetite, diarrhea, but no running nose, nasal obstruction, dyspnea, abnormal smell and taste. The infection rapidly developed from medium to critical. On the basis of standard treatment, Delta variant CP was intravenous dripped on the 10th day of hospital admission (the 6th day after becoming severe). The patient's condition improved rapidly.Conclusion:The curative efficacy evaluation of this patient proved that CP therapy is of great value in the treatment of severe and critical COVID-19.

Objective:To compare the diagnostic accuracy between multiparametric magnetic resonance imaging (mp-MRI) and biparametric magnetic resonance imaging (bp-MRI) in muscle-invasive bladder cancer (MIBC).Methods:The clinical data of 195 patients with bladder cancer at the First Affiliated Hospital of Nanjing Medical University from July 2020 to June 2022, were retrospectively reviewed. There were 160 males and 35 females, with the median age of 68(61, 76)years old. Mp-MRI was performed on each patient within 6 weeks before transurethral resection of bladder tumor or radical cystectomy. Each patients’ images were divided into two sets. Set 1 (bp-MRI) included the axial, sagittal, coronal T2-weighted images (T2WI), and axial diffusion-weighted images (DWI) or apparent diffusion coefficient maps. Set 2 (mp-MRI) included Set 1 images in addition to dynamic contrast-enhanced images. All images were independently reviewed and evaluated by two radiologists. Mp-MRI was evaluated according to the Vesical Imaging-Reporting and Data System (VI-RADS)guideline, and bp-MRI was evaluated according to two types of criteria. Bp-MRI (Criterion A): VI-RADS scoring is determined 2 when T2WI 3-point with DWI 2-point. Bp-MRI (Criterion B): VI-RADS scoring is determined 3 when T2WI 3-point with DWI 2-point. VI-RADS scoring ≥ 3 or ≥ 4 was used as the cut-off value to predict MIBC. The sensitivity, specificity, positive predictive value, and negative predictive value of mp-MRI, bp-MRI (Criterion A), and bp-MRI (Criterion B) were calculated, as well as receiver operating characteristic curves and the areas under the curve (AUC).Results:Of 195 patients, 135 patients (69.2%) were pathologically confirmed as NMIBC and 60 patients (30.8%) were MIBC. When the VI-RADS cut-off value was ≥ 3, the sensitivity of mp-MRI, bp-MRI (Criterion A), and bp-MRI (Criterion B) were identical, all at 88.3% (53/60). The specificity of bp-MRI (Criterion A), bp-MRI (Criterion B), and mp-MRI were 88.9% (120/135), 73.3% (99/13), and 86.7% (117/135), respectively. When the VI-RADS cut-off value was ≥ 4, both bp-MRI (Criterion A) and bp-MRI (Criterion B) were classified as the same criterion. The sensitivity of bp-MRI and mp-MRI were 70.0% (42/60) and 75.0% (45/60), respectively. The specificity of bp-MRI and mp-MRI were identical, at 95.6% (129/135). The AUC for bp-MRI (Criterion A), bp-MRI (Criterion B), and mp-MRI were 0.927 (95% CI 0.881-0.959), 0.904 (95% CI 0.853-0.941), and 0.927 (95% CI 0.881-0.959), respectively. The AUC for bp-MRI (Criterion A) and mp-MRI were significantly higher than that of bp-MRI (Criterion B) ( P<0.001). There was no significant difference in AUC between bp-MRI (Criterion A) and mp-MRI ( P=0.939). Conclusions:Bp-MRI (Criterion A), VI-RADS scoring is determined 2 when T2WI 3-point with DWI 2-point, shows comparable diagnostic accuracy in predicting MIBC with mp-MRI. Compared to bp-MRI (Criterion B), the corresponding situation when VI-RADS scoring is determined 3, bp-MRI (Criterion A) may have better diagnostic accuracy than bp-MRI (Criterion B) in predicting MIBC.

Objective To compare the efficacy of retroperitoneal robot-assisted laparoscopic partial nephrectomy(RALPN)in the treatment of ventral and dorsal renal tumor.Methods The clinical data of 131 patients with renal tumor who underwent retroperitoneal RALPN at our hospital during Jan.2021 and Feb.2024 were retrospectively analyzed.The patients were divided into the ventral renal tumor group(n=35)and dorsal renal tumor group(n=96)according to preoperative images.Perioperative outcomes and prognosis were compared between the two groups.Results All 131 surgeries were successfully completed without conversion to open surgery or abdominal organ injury.There were no significant differences in warm ischemia time[median(interquartile range,IQR):20(16.75)min vs.22(IQR:15.25)min],operation time[57.0(IQR:29.5)min vs.58.5(IQR:39.75)min],estimated intraoperative blood loss[50(IQR:80)mL vs.50(IQR:80)mL],proportion of patients with intraoperative blood transfusion(0 vs.2.1％),proportion of patients using four arms during operation(42.9％vs.37.5％),postoperative reduction of hemoglobin(Hb)[(-12.9±9.0)g/L vs.(-11.5±9.4)g/L],reduction of estimated glomerular filtration rate(eGFR)[(-3.8±12.4)mL/min vs.(-7.0±13.6)mL/min],postoperative hospital stay[4(IQR:0)d vs.4(IQR:2)d],and proportion of meeting"Trifecta"criteria(94.3％vs.86.5％,P＞0.05).During the median follow-up of 9.8(5.5,24.0)months,no death occurred,and no recurrence or metastasis were observed except that bone metastasis occurred in 1 patient in the dorsal renal tumor group.Conclusion Both ventral and dorsal renal tumors can be treated with retroperitoneal RALPN,and the surgical outcomes of both are comparable.

Objective:To discuss the feasibility and safety of robot-assisted laparoscopic retroperitoneal tumor resection in prone position.Methods:From August 2023 to January 2024, a total of eight patients with retroperitoneal tumors from the First Affiliated Hospital of Nanjing Medical University were retrospectively analyzed, including two males and six females. The average age was (47.4±12.5) years, average BMI was (24.4±3.5) kg/m 2 and median ASA grade was 2(2, 3). Retroperitoneal tumors were identified preoperatively through CT or MRI. The imaging revealed 4 cases of adrenal tumors located on the left side, 2 on the right side, and 2 non-adrenal tumors situated on the left side. The preoperative diagnoses included 2 cases of non-functional adrenal tumors, 2 cases of pheochromocytoma, 1 case of Cushing’s syndrome, 1 case of metastatic renal cell carcinoma, and 2 cases of non-adrenal tumors. Robot-assisted laparoscopic retroperitoneal tumor resection was performed with all patients in prone position. The inferior margin of the fourth lumbar vertebra (L4) was determined by the line connecting the highest points of the iliac crests bilaterally. Subsequently, the inferior margins of the L1-L3 vertebrae were sequentially identified. The surgical field was then divided into three equal segments, utilizing the posterior midline of the spine and the midaxillary line as boundaries. The medial division was situated approximately at the lateral border of the vertical spinal muscles, while the lateral division was placed near the tip of the 12th rib. A longitudinal incision of approximately 3 cm in length was created within the lateral division between L2 and L3 for the insertion of a camera trocar. The extraperitoneal space was subsequently dilated using a self-made balloon, and two 8 mm trocars were placed as operative ports along the medial division and the midaxillary line, respectively, under finger guidance. Assistance trocars, one or two 12 mm in diameter, were introduced above the level of the iliac crest. During the operation, the extraperitoneal adipose tissue was removed and the Gerota's fascia was opened. For non-adrenal retroperitoneal tumours, the major blood vessels around the tumour were suspended and fixed, by titanium clips or Hem-o-lok clips to dissect the small arteries and veins, and the tumour was carefully isolated and completely resected. For adrenal tumours, the fat capsule around the upper pole of the kidney were removed, the adrenal gland was exposed, and then the tumour was removed completely along its capsule. If total adrenalectomy is performed, the central adrenal vein was clamped and dissected. The periphery of the adrenal gland was gradually dissected until the adrenal gland was completely removed.The perioperative data, including patient positioning time, trocar placement time, operation time, intraoperative blood loss, postoperative complications, postoperative hospital stay, and postoperative drainage tube removal time, as well as recurrence and metastasis, were recorded.Two patients underwent partial nephrectomy due to renal tumor, and only the time for retroperitoneal tumor resection was included in calculating operation time. Results:All 8 surgeries were successfully completed without dramatic blood pressure fluctuations.There was no conversion to open surgery or abdominal organ injury. The patient positioning time was (5.1±0.4) minutes, trocar placement time was (16.6±1.3) minutes, operation time was (28.8±13.8) minutes, intraoperative blood loss was (65.0±28.7) ml, postoperative hospital stay was (3.6±0.9) days, and drainage tube removal time was (2.8±1.0) days. No intraoperative or postoperative blood transfusions were required. Postoperatively diagnosed pathologies included: 2 cases of adrenal pheochromocytoma, 2 cases of adrenal sebaceous adenoma, 2 cases of retroperitoneal schwannoma, 1 case of adrenal myelolipoma, and 1 case of adrenal metastatic renal cell carcinoma. The average tumor size for all patients was (4.3±1.5) cm. After a follow-up of 2.0-7.2 months, there were no recorded postoperative complications, including haemorrhage, infections, acute hypotension, or adrenocortical insufficiency. Additionally, no evidence of tumor recurrence or metastasis was observed up during foolow-up.Conclusions:Robot-assisted laparoscopic retroperitoneal tumor resection in prone position could be a safe and feasible surgical approach with short operative time, low bleeding, and fast postoperative recovery.

Aspergillosis ; Humans ; Mucor ; Mucormycosis/drug therapy*