Objective To investigate the gene therapy of hypoparathyroidism (HPT) with hemopoietic stem cell s(HSCs) integrated with parathyroid hormone (PTH) gene.Methods Human PTH gene was amplified by polymerase chain reaction from pcDNA 3.1 -PTH templates and inserted into murine stem cell virus (MSCV) vectors. The plasmids recombinated with PTH gene were transfected into PA317 packaging cells and screened for positive clones with Geneticin. The condensed recombinant retroviruses were obtained and used to infect HSCs coming from umbilical cord blood. The infected HSCs were injected into the mice suffering from HPT. The change of symptoms and serum level of PTH and calcium in each group of mice was observed.Results Human PTH gene was proved to be inserted into MSCV vectors and the titres were up to 2?10 7 colony forming unit (CFU)/ml in condensed retroviral solution. The secretion of PTH reached 15 ng per 10 6 cells per 48 h. The wild type viruses were not detected by PCR amplification. The mice suffering from HPT recovered quickly and the serum level of calcium and PTH remained normal for about 3 months after the HSCs recombinated with PTH gene were injected into them. In the groups of mice subject to the injection of the recombinant retroviruses containing PTH gene, the serum level of PTH and Calcium was significantly increased in a short time, and then declined slowly. And the mice took on the manifestation of hypoparathyroidism again. The mice only subject to the implantation of HSCs were all dead within 20 days.Conclusion A long term therapeutic effect could be obtained by infusing the HSCs which were integrated with PTH gene into the mice suffered from hypoparathyroidism.

Taxus canadensis distributed mainly in North America, such as northern Minnesota, Newfoundland, south to Wisconsin and Pennsylvania. Its composition has been shown to be very different from other species, and in recent years, some new skeletons also have been found in Canada yew. Through analysis of the taxanes content on various Taxus plants containing taxanes, the results showed a higher content of taxol in T. canadensis. Based on the current research on T. canadensis (from the research results of the author in recent years, as well as from studies of scholars in the field), the paper outlined the research progress in recent years on the chemical constituents of taxane of T. canadensis and the spectral characteristics of various types of compounds. Besides, this paper analyzed the present research about solutions for the taxol drug source crisis.
Animals ; Bridged-Ring Compounds ; chemical synthesis ; chemistry ; pharmacology ; Humans ; Molecular Structure ; Plant Extracts ; chemical synthesis ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry ; Taxoids ; chemical synthesis ; chemistry ; pharmacology ; Taxus ; chemistry

Objective To investigate factors influencing islet isolation and purification in dogs and to explore how to obtain massive purified viable islets from canine pancreas.To provide experimental evidence for pancreatic islet transplantation in clinical practice.Methods Islets were isolated from pancreas of 22 dogs by modified automated techniques,followed by purification using continuous density gradients.The purity of islets was determined by dithizone (DTZ) staining,and the function of islets was evaluated by glucose stimulating insulin release test.Results The crude islets yield was ( 155 040 ±310) IEQ/pancreas while the yield was (74 200 ±185) IEQ/pancreas after purification.The average purity was (89.4 ±2.6) ％.The recovery rate was(47.8 ±1.3) ％.Islet viability was(93.0 ± 1.7)％.The modified techniques improved both the yield and the quality in islet isolation and purification.The islets were morphologically intact after purification.Glucose stimulating insulin release test showed that the difference of the secreted insulin concentration had statistical significance ( P ＜0.001 ) between low concentration glucose group and high concentration glucose group.Conclusion It is of great value for clinical research to obtain enough islets through islet isolation and purification in dogs.

Objective To study an improved automated method for isolation and purification of large amounts of human pancreatic islets from large mammals,and try to create conditions for preparation and isolation of large amounts of human islets.Methods An improved automated system was used to isolate and purify panreatic islets of sogs.Under the general anaesthesia(GA) condition,the pancreas of the dogs was via in situ vascular perfused using cold HC-A solution and then removed en bloc with duodenum and spleen.Then they were placed in cold UW aolutian.Intraductal collagenase-Ⅴ and pefabloc(4 ℃) was delivered with controlled perfusion.lslets were dissociated in system of Ricordi Chamber and purified islets separated with Ficoll density gradient centrifugation under controlled temperature.Digestion time,islets remaining trapped in exocrine tissue,final islet purity,insulin and C-pep secretory activity,and islet recovery were observed.The purified islets were observed under light microscope and electronic microscope after 24 h culture.Results The digestion time rate was(25.0?6.0) min,islets remaining trapped in exocrine tissue was(9.4?2.4)%,final islet purify rate was(89.7?3.5)%,islet recovery after digestion was(17.2?3.6)?104 IEQ/pancreas.Islet recovery after purification was(8.3?2.0)?104IEQ/pancreas.Insulin and C-pep secretory activity of the purified islets and their ultrastructure after 24 h culture were ideal.Conclusions Our improved automated method and facilities for isolation of canine pancreatic islets are reliable,The morphology and function of the procured islets are excellent and they can foreseeably be used for large-scale preparation of huma islets for clinical use.

OBJECTIVETo investigate the factors influencing the prognosis of penis-sparing surgery (PSS) for early-stage penile cancer.

METHODSWe retrospectively studied the clinical data about 45 cases of early-stage penile cancer treated by PSS from January 2007 to December 2014. We calculated the rate of local recurrence-free survival by the Kaplan-Meier method, and conducted univariate and multivariate COX regression analyses on the relevant factors including the patient's age, marital status, tumor location, tumor size, postoperative sexual life, histological grade, and TNM stage.

RESULTSOne-year and three-year local recurrence-free survival rates were 95.5% and 52.2%, respectively. Multivariate analysis demonstrated that the histological grade (P = 0.039) and postoperative sexual life (P = 0.049) were independent factors for the prognosis of PSS. Logistic regression showed the patients age to be significantly associated with histological grade (P = 0.014).

CONCLUSIONHistological grade and postoperative sexual life are important independent prognostic factors of PSS for early-stage penile cancer, and the patients age is associated with the prognosis of PSS through its influence on the tumor grade.

Age Factors ; Disease-Free Survival ; Humans ; Kaplan-Meier Estimate ; Logistic Models ; Male ; Multivariate Analysis ; Neoplasm Grading ; Organ Sparing Treatments ; Penile Neoplasms ; surgery ; Penis ; surgery ; Prognosis ; Proportional Hazards Models ; Quality of Life ; Retrospective Studies

OBJECTIVETo explore the analgesic effect of Zhixin Formula (ZF) and its effects on spinal glial fibrillary acidic protein (glial fibillary acidic protein, GFAP, a marker of astrocyte) , CD11b (a maker of microglia), Toll like receptors (TLR2 and TLR4) and nuclear factor κB (NF-κB) in bone cancer pain model rats.

METHODSTotally 20 male SD rats were randomly divided into the blank control group and the bone cancer pain group, 10 in each group. The bone cancer pain model was induced by injecting ascites tumor fluid containing 3 x 10(3) Walker256 cell line from the left tibia. Ethological tests, X-ray test, and HE staining were performed to confirm a successful modeling. After model was successfully established, 70 male SD rats were randomly divided into seven groups, 10 in each group: the blank control group, the bone cancer pain group (as the model group), the Western medicine (WM) group (Tramadol Hydrochloride), the high dose ZF group, the middle dose ZF group, the low dose ZF group, and the Chinese medicine (CM) group (Wulin Zhitong Capsule). Fourteen days after modeling, rats in the high, middle, and low dose ZF groups were administrated by gastrogavage with 9, 4.5, and 2.25 g/kg ZF water condensed preparation respectively, once a day for seven consecutive days. On day 21 MS typical protein expressions including GFAP, CD11b, TLR (2,4) and NF-κB from cornu dorsal medullae spindis L4-L5 were detected by immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.

RESULTSCompare with the blank control group, increased weight in the model group was slow and showed a decreasing trend (P < 0.01), spontaneous ambulatory pain score (SAPS) obviously increased (P < 0.01), paw withdrawal threshold (PWT) obviously decreased in the model group (P < 0.05, P < 0.01). Results of lateral tibial X-ray and HE staining showed obvious changes and damage occurred in bone structures of rats in the model group. Immunohistochemistry showed that GFAP expression significantly increased in the model group (P < 0.05). Protein levels of NF-κB also significantly increased in the model group (P < 0.05). Compared with the model group, CD11b expressions obviously decreased in the middle and high dose ZF groups (P < 0.01). Meanwhile, protein expressions of TLR2 and TLR4, as well as NF-κB also obviously decreased (P < 0.05).

CONCLUSIONZF had analgesic effect, which might be probably related to inhibiting proliferation and activation of gliocytes, as well as activation of TLRs and NF-κB.

Animals ; Astrocytes ; Bone Neoplasms ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Glial Fibrillary Acidic Protein ; Male ; Microglia ; NF-kappa B ; metabolism ; Osteosarcoma ; Pain ; Rats ; Rats, Sprague-Dawley

Objective In view of being short of the mammalian model in neuroma-in-continuity,the experiment injured the part of peroneal nerve to the formation of the neuroma-in-continuity and was applied to the foundation of farther research.Methods Twelve New Zeland rabbits were selected as experimental sub- jects randomly.One lateral peroneal nerves of twelve rabbits were resected,the damaged nervous tissues' slice were showed to the typical pathological changes of neuroma by the stain of HE,luxol fast blue after six weeks. As compared with the health sides of six model rabbits,the methods of real-time PCR and Western blot were used to evaluate the expression of CNTF,CGRP mRNA and protein in injured nerves and L_7、S_1 dorsal root ganglions respectively.Results The injured nerve formed the typical pathological changes of neuroma at six weeks.Compared with hea|thg side the expression of CNTF mRNA and protein was down-graded at the lateral of neuroma(P＜0.05),and the expression of CGRP mRNA and protein was up-graded(P＜0.05).Con- clusion The method of partly injuring the peroneal nerve could effectively set up the model of the neuroma-in- continuity,furthermore,resulted to the expression changes of the CNTF,CGRP mRNA and protein.

BACKGROUND: The study aimed to explore the effects of hypothermia state induced by 4 oC normal saline (NS) on liver biochemistry, enzymology and morphology after restoration of spontaneous circulation (ROSC) by cardiopulmonary resuscitation (CPR) in swine. METHODS: After 4 minutes of ventricular fibrillation (VF), standard CPR was carried out. Then the survivors were divided into two groups: low temperature group and normal temperature group. The low temperature (LT) group (n=5) received continuously 4 oC NS at the speed of 1.33 mL/kg per minute for 22 minutes, then at the speed lowering to 10 mL/kg per hour. The normal temperature (NT) group (n=5) received NS with normal room temperature at the same speed of the LT group. Hemodynamic status and oxygen metabolism were monitored and the levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were measured in blood samples obtained at baseline and at 10 minutes, 2 hours and 4 hours after ROSC. At 24 hours after ROSC, the animals were killed and the liver was removed to determine the Na+-K+-ATPase and Ca2+-ATPase enzyme activities and histological changes under a light or electron microscope. RESULTS: Core temperature was decreased in the LT group (P<0.05), while HR, MAP and CPP were not significantly decreased (P>0.05) compared with the NT group (P>0.05). The oxygen extraction ratio was lower in the LT group than in the NT group (P<0.05). The serum levels of ALT, AST and LDH increased in both groups but not significantly in the LT group. The enzyme activity of liver ATP was much higher in the LT group (Na+-K+-ATP enzyme: 8.64±3.32 U vs. 3.28±0.71 U; Ca2+-ATP enzyme: 10.92±2.12 U vs. 2.75±0.78 U, P<0.05). The LT group showed less cellular edema, inflammation and few damaged mitochondria as compared with the NT group. CONCLUSION: These data suggested that infusing 4 oC NS continuously after ROSC could quickly lower the core body temperature, while maintaining a stable hemodynamic state and balancing oxygen metabolism, which protect the liver in terms of biochemistry, enzymology and histology after CPR.

Biopsy ; Biopsy, Needle ; Bone Marrow ; pathology ; Bone Marrow Examination ; methods ; Bone Marrow Neoplasms ; pathology ; secondary ; Breast Neoplasms ; pathology ; Cytological Techniques ; Female ; Humans ; Lung Neoplasms ; pathology ; Male ; Prostatic Neoplasms ; pathology ; Retrospective Studies ; Stomach Neoplasms ; pathology

The infant (a girl aged 6 months) was admitted to the hospital because of oliguria and acute renal dysfunction. The laboratory examination results showed serious metabolic acidosis and increased blood urea nitrogen and serum creatinine levels. The patient continued to be anuric after 10 days of treatment with continuous renal replacement therapy (CRRT). she died a day later. The family history showed that the patient's sister died of acute renal failure 6 months after birth. The genomic sequencing results showed AGXT mutation in the patient and confirmed the diagnosis of primary hyperoxaluria type 1 (PH1). Her parents were heterozygous carriers. PH1 should be considered when the children have abnormal renal function or recurrent renal calculi or have a family history of these symptoms. AGXT gene analysis is an important method for PH1 diagnosis.
Acute Kidney Injury ; etiology ; Female ; Humans ; Hyperoxaluria, Primary ; complications ; Infant ; Mutation ; Oliguria ; etiology ; Transaminases ; genetics