Objective To clone NT-3 gene from normal rat brain and to purify its fusion protein and to prepare specific high titer antibody so that to provide a foundation for further study for peripheral nerve injury.MethodsWe amplified target gene by RT-PCR and cloned it into the vector of pMD-18T,then analyzed its sequence and compared it with the sequence from GenBank.We subcloned it into pRSET-A vector and introduced it into Escherichia coli BL21.The expression was induced by IPTG,and identified by SDS-PAGE.The fusion protein was purified by niccolum purify kit.We immuned rabbits with immunological adjuvant for specificity antibody preparation.Results We got a 777 bp gene segment by RT-PCR.The DNA sequence was identical to rat NT-3 gene sequence in GenBank.It proved that the target gene was correctly inserted into the vector.A new protein band of about 34 ku appeared on SDS-PAGE after induction of IPTG.A specific high titer antibody of 1∶64000 was gained by immunizing rabbits with adjuvant.

Objective To investigate the urinary endothelin-1 (ET-1) excretion and urinary sodium excretion,microalbuminuria and ambulatory blood pressure(ABP) in salt-sensitive(SS) hypertension patients. Methods Twenty-one cases of normotensive subjects and 32 cases of uncomplicated hypertensive patients were recruited in this study. Salt sensitivity was determined by acute venous saline loading test. Before saline loading, 24-hour ABP measurements were performed. Urine samples were collected to assay ET-1 ,urinary sodium excretion and urinary albumin excretion(UAF). Results Compared to slat-resistant(SR) subgroup, SS showed low urinary ET-1 excretion in normotensive group (P＜0.05) or hypertensive group (P＜0.01) ,regardless of saline loading or not. The nighttime MAP of SS was higher than SR subgroup in normotensive or hypertensive group. Urinary sodium excretion during 4h of saline loading was significantly lower in SS than that in SR hypertensive patients (P＜0. 05). Twenty-four-hour UAE of SS patients was higher than SR group (P＜0.01). Results of further correlation analysis indicated that the urinary ET-1 excretion was positively related to urinary sodium content and negatively to ABP and UAE. Conclusion Urinary ET-1 is low in SS normotensives or hypertension patients,which may play a role in renal sodium retention and renal impairment of SS hypertension patients.

Objective:To study the detection value of platelet activation indexes and platelet parameters in patients with hepatitis B-induced cirrhosis.Methods:40 patients with hepatitis B-induced cirrhosis in our hospital from November 2015 to August 2016 were selected as the observation group.40 healthy subjects went through physical examination during the same period were chosen as the control group.The platelet activation indexes and platelet parameters of subjects with same age in two groups were detected and compared.The detection results of the two items in patients with different stages of hepatitis B-induced cirrhosis were compared,too.Results:The PLT of observation group was lower than that of control group,while the other platelet parameters and platelet activation indexes were all higher than those of control group.The detection results of two items in patients with different stages of hepatitis B-induced cirrhosis were with significant differences,all P＜0.05.Conclusion:The platelet activation indexes and platelet parameters in patients with hepatitis B-induced cirrhosis are obviously abnormally expressed.There are significant differences among the detection results in patients with different stages.Therefore,surveillance of those indexes in patients with hepatitis B-induced cirrhosis should be strengthened.

BACKGROUND:It has been studied that the distribution of bone morphogenetic protein 2 is regular under bone defect situation. OBJECTIVE:To observe the expression of bone morphogenetic protein 2 in rabbit radial defect site with different lengths. METHODS:Forty-eight New Zealand rabbits were divided into two groups randomly, 0.5 cm bone defect and 3.0 cm bone defect were made by wire saw at the middle part of radius bone after anaesthesia. RESULTS AND CONCLUSION:Western blot results showed that in the 0.5 cm bone defect group, the expression of bone morphogenetic protein 2 of the tissues in the bone defect site was increased gradual y at 1, 3, 4 weeks after operation, and the expression in each defect group was increased when compared with that immediately after injury (P<0.05). In the 3.0 cm bone defect group, the expression of bone morphogenetic protein 2 of tissues in bone defect site was increased gradual y and reached to its peak at 3 weeks after the operation (P<0.05), and the peak value in the 3.0 cm bone defect group was significantly higher than that in 0.5 cm bone defect group (P<0.05). The peak value was maintained in high level. The comparison of bone cal us formation showed that the bone cal us formation of 3.0 cm bone defect group was less than that of the 0.5 cm bone defect group at 3 and 4 weeks after operation (P<0.05). The results indicate that expression of the bone morphogenetic protein 2 in 3.0 cm bone defect site is increased significantly, but the expression level cannot make the bone defect heal itself.

OBJECTIVETo evaluate the hemostatic role of hemocoagulase in abdominal operation and its effects on coagulation.

METHODS180 patients receiving abdominal operation were studied prospectively by randomized, double-blind controlled and multicenter design. They were divided into Hemocoagulase group (60 patients), lizhixue group (60), and manitol hexanitrate group (60). The groups were, observed in terms of the effects on hemostatic time, hemorrhagic volume, hemorrhagic volume per square unit, and body coagulation (BT, CT, PT, APTT and PLT) parameters.

RESULTSThe groups received different drugs. The average hemostatic time in the hemocoagulase group was 121.6 s, hemorrhagic volume was 9.6 g, and hemorrhagic volume per square unit was 0.2 g. The similar results were observed in the lizhixue group (P > 0.05), but they were significantly different (P < 0.05) from those of the manitol hexanitrate group (159.2 s, 12.49 g, 0.3 g). In the hemocoagulase and lizhixue groups hemorrhagic and hemoagglution time decreased 30 minutes and 1 day after operation. This finding was significantly differenct from that in the manitol hexanitrate group (P < 0.05).

CONCLUSIONHemocoagulase plays a good hemostatic role in the hemorrhagic capillary at abdominal incision.

Abdomen ; Adolescent ; Adult ; Aged ; Batroxobin ; adverse effects ; pharmacology ; therapeutic use ; Blood Coagulation ; drug effects ; Double-Blind Method ; Female ; Hemostasis, Surgical ; methods ; Hemostatics ; therapeutic use ; Humans ; Male ; Middle Aged ; Prospective Studies

OBJECTIVE：To investigate the effects of metformin on malignant phenotype of pancreatic cancer BxPC- 3 cells. METHODS：Using human pancreatic cancer BxPC- 3 cells with natural deletion of Smad4 gene as reaserch objects ，CCK-8 assay and flow cytometry were used to detect the proliferation and apoptosis of BxPC- 3 cells after treated with different doses of metformin（5，10，20 mmol/L）for 24 h. The cell survival rate and apoptosis rate were calculated. Transwell assay was used to test the migration of cells after treated with different doses of metformin （10，20 mmol/L）for 24 h. The number of migrating cells was recorded. qRT-PCR and Western blotting assay were performed to determine mRNA and protein expression of E-cadherin ，Vimentin and RGC- 32 in cells. RESULTS ：Compared with control group and 5 mmol/L metformin group ，survival rate of cells were decreased significantly in 10，20 mmol/L metformin groups ，while apoptosis rate was increased significantly ；the apoptosis rate in 20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group （P＜0.05）. Compared with control group ， the number of migrating cells was decreased significantly in 10，20 mmol/L metformin groups ，and the 20 mmol/L metformin group was significantly lower than 10 mmol/L metformin group （P＜0.05）. Relative mRNA and protein expression of E-cadherin were increased significantly in 10，20 mmol/L metformin groups ，and relative mRNA expression of E-cadherin in 20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group. Relative mRNA expression of Vimentin in 10 mmol/L metformin group ，relative mRNA and protein expression of Vimentin in 20 mmol/L metformin group ，relative mRNA and protein expression of RGC- 32 in 10，20 mmol/L metformin groups were decreased significantly ；relative mRNA and protein expression of Vimentin as well as mRNA expression of RGC- 32 in 20 mmol/L metformin group were significantly lower than 10 mmol/L metformin group （P＜0.05 or P＜0.01）. CONCLUSIONS ：Metformin can inhibit the proliferation and migration of pancreatic cancer cells through smael-independent pathways in a dose- dependent manner ，and promote their apoptosis ，which is associated with the inhibition epithelial- mesenchymal transition and the expression of RGC- 32 of pancreatic cancer.

Objective To evaluate the clinical value of holographic image navigation in urological laparoscopic and robotic surgery.Methods The data of patients were reviewed retrospectively for whom accepted holographic image navigation laparoscopic and robotic surgery from Jan.2019 to Dec.2019 in Beijing United Family Hospital and other 18 medical centers,including 78 cases of renal tumor,2 cases of bladder cancer,2 cases of adrenal gland tumor,1 cases of renal cyst,1 case of prostate cancer,1 case of sweat gland carcinoma with lymph node metastasis,1 case of pelvic metastasis after radical cystectomy.All the patients underwent operations.In the laparoscopic surgery group,there were 27 cases of partial nephrectomy,1 case of radical prostatectomy,2 cases of radical cystectomy and 2 cases of adrenalectomy.In the da Vinci robotic surgery group of 54 cases,there were 51 cases of partial nephrectomy,1 case of retroperitoneal lymph node dissection,1 case of retroperitoneal bilateral renal cyst deroofing and 1 case of resection of pelvic metastasis.There were 41 partial nephrectomy patients with available clinical data for statistic,with a median age of 53.5 years (range 24-76),including 26 males and 15 females.The median R.E.N.A.L score was 7.8 (range 4-11).Before the operation,the engineers established the holographic image based on the contrast CT images and reports.The surgeon applied the holographic image for preoperative planning.During the operation,the navigation was achieved by real time fusing holographic images with the laparoscopic surgery images in the screen.Results All the procedures had been complete uneventfully.The holographic images helped surgeon in understanding the visual three-dimension structure and relation of vessels supplying tumor or resection tissue,lymph nodes and nerves.By manipulating the holographic images extracorporeally,the fused image guide surgeons about location vessel,lymph node and other important structure and then facilitate the delicate dissection.For the 41 cases with available clinical data including 23 cases of robotic-assisted partial nephrectomy and 18 cases of laparoscopic nephrectomy,the median operation time was 140 (range 50-225) min,the median warm ischemia time was 23 (range 14-60) min,the median blood loss was 80(range 5-1 200) ml.In the robotic surgery group,the median operation time was 140 (range 50-215)min,the median warm i schemia time was 21 (range 17-40)min,the median blood loss was 150(range 30-1 200)ml.In the laparoscopic surgery group,the median operation time was 160(range 80-225)min,the median warm ischemia time was 25 (range 14-60)min,the median blood loss was 50 (range 5-1 200) ml.All the patients had no adjacent organ injury during operation.There were 2 cases with Clavien Ⅱ complications.One required transfusion and the other one suffered hematoma post-operation.However,the tumors were located in the renal hilus for these 2 cases and the R.E.N.A.L scores were both 11.Conclusions Holographic image navigation can help location and recognize important anatomic structures during the surgical procedures..This technique will reduce the tissue injury,decrease the complications and improve the success rate of surgery.

Objective: To investigate and analyze the clinical features, epidemiologic information and pathogenic characteristics of a rabies patient. Methods: Clinical data of the patient(boy) was collected and epidemiological survey was conducted, fluorescence quantitative reverse transcription-polymerase chain reaction (FQRT-PCR) and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the samples of saliva, cerebrospinal fluid (CSF), skin tissue with hair follicle at the back of the neck for rabies laboratory diagnosis. Results: Early symptoms of the boy were vomiting, diarrhea, fever and irritability, followed by coma and death. The boy had nasal trauma one month ago and the domestic dog died of illness during the same period. He did not accept the rabies post-exposure prophylaxis (PEP). The result of the saliva sample was positive by FQRT-PCR. The predicted segments of the glycoprotein(G), nucleoprotein (N) genes of rabies virus were amplified from the positive saliva sample of the patient by RT-PCR. Compared with rabies virus strains in Henan province, the nucleotide homology and amino acid homology in G gene segment were 96.5%-98.8% and 96.5%-99.2% respectively. Conclusions The case was diagnosed in laboratory as rabies case. The pathogenic rabies virus strain was endemic in Henan province. The nasal trauma, the dead domestic dog were probably related to the infection of the boy.

【Objective】 To establish a blood quality monitoring indicator system, in order to continuously improve blood quality and standardized management. 【Methods】 Based on the research of literature and standards, and guided by the key control points of blood collection and supply process, the blood quality monitoring indicator system was developed. Through two rounds of Delphi expert consultation, the indicator content was further revised and improved according to expert opinions after six months of trial implementation. The indicator weight was calculated by questionnaire and analytic hierarchy process. 【Results】 A blood quality monitoring indicator system covering the whole process of blood collection and supply was constructed, including five primary indicators, namely blood donation service, blood component preparation, blood testing, blood supply and quality control, as well as 72 secondary indicators, including definitions, calculation formulas, etc. Two rounds of expert consultation and two rounds of feasibility study meeting were held to revise 17 items and the weight of each indicator was obtained through the analytic hierarchy process. After partial adjustments, a blood quality monitoring indicator system was formed. 【Conclusion】 A blood quality monitoring indicator system covering the whole process of blood collection and supply has been established for the first time, which can effectively evaluate the quality management level of blood banks and coordinate blood quality control activities of blood banks in Shandong like pieces in a chess game, thus improving the standardized management level

With the increasing proportion of human papilloma virus(HPV)infection in the pathogenic factors of oro-pharyngeal cancer,a series of changes have occurred in the surgical treatment.While the treatment mode has been im-proved,there are still many problems,including the inconsistency between diagnosis and treatment modes,the lack of popularization of reconstruction technology,the imperfect post-treatment rehabilitation system,and the lack of effective preventive measures.Especially in terms of treatment mode for early oropharyngeal cancer,there is no unified conclu-sion whether it is surgery alone or radiotherapy alone,and whether robotic minimally invasive surgery has better func-tional protection than radiotherapy.For advanced oropharyngeal cancer,there is greater controversy over the treatment mode.It is still unclear whether to adopt a non-surgical treatment mode of synchronous chemoradiotherapy or induction chemotherapy combined with synchronous chemoradiotherapy,or a treatment mode of surgery combined with postopera-tive chemoradiotherapy.In order to standardize the surgical treatment of oropharyngeal cancer in China and clarify the indications for surgical treatment of oropharyngeal cancer,this expert consensus,based on the characteristics and treat-ment status of oropharyngeal cancer in China and combined with the international latest theories and practices,forms consensus opinions in multiple aspects of preoperative evaluation,surgical indication determination,primary tumor re-section,neck lymph node dissection,postoperative defect repair,postoperative complication management prognosis and follow-up of oropharyngeal cancer patients.The key points include:① Before the treatment of oropharyngeal cancer,the expression of P16 protein should be detected to clarify HPV status;② Perform enhanced magnetic resonance imaging of the maxillofacial region before surgery to evaluate the invasion of oropharyngeal cancer and guide precise surgical resec-tion of oropharyngeal cancer.Evaluating mouth opening and airway status is crucial for surgical approach decisions and postoperative risk prediction;③ For oropharyngeal cancer patients who have to undergo major surgery and cannot eat for one to two months,it is recommended to undergo percutaneous endoscopic gastrostomy before surgery to effectively improve their nutritional intake during treatment;④ Early-stage oropharyngeal cancer patients may opt for either sur-gery alone or radiation therapy alone.For intermediate and advanced stages,HPV-related oropharyngeal cancer general-ly prioritizes radiation therapy,with concurrent chemotherapy considered based on tumor staging.Surgical treatment is recommended as the first choice for HPV unrelated oropharyngeal squamous cell carcinoma(including primary and re-current)and recurrent HPV related oropharyngeal squamous cell carcinoma after radiotherapy and chemotherapy;⑤ For primary exogenous T1-2 oropharyngeal cancer,direct surgery through the oral approach or da Vinci robotic sur-gery is preferred.For T3-4 patients with advanced oropharyngeal cancer,it is recommended to use temporary mandibu-lectomy approach and lateral pharyngotomy approach for surgery as appropriate;⑥ For cT1-2N0 oropharyngeal cancer patients with tumor invasion depth＞3 mm and cT3-4N0 HPV unrelated oropharyngeal cancer patients,selective neck dissection of levels ⅠB to Ⅳ is recommended.For cN+HPV unrelated oropharyngeal cancer patients,therapeutic neck dissection in regions Ⅰ-Ⅴ is advised;⑦ If PET-CT scan at 12 or more weeks after completion of radiation shows intense FDG uptake in any node,or imaging suggests continuous enlargement of lymph nodes,the patient should undergo neck dissection;⑧ For patients with suspected extracapsular invasion preoperatively,lymph node dissection should include removal of surrounding muscle and adipose connective tissue;⑨ The reconstruction of oropharyngeal cancer defects should follow the principle of reconstruction steps,with priority given to adjacent flaps,followed by distal pedicled flaps,and finally free flaps.The anterolateral thigh flap with abundant tissue can be used as the preferred flap for large-scale postoperative defects.