Objective To investigate the expression of Clq complement receptor(ClqRp and gClqR)in the peripheral blood mononuclear cells(PBMC)of systemic lupus erythematosus(SLE), and the corre|ation between serum levels of complement lq(Clq)and anti-Clq autoantibodies(ClqAb)with ClqRp and gClqR is analyzed. The probable mechanism of ClqRp and gClqR in the development of SLE is explored. Methods The peripherial blood monouclear cells of 58 SLE patients and 30 healthy subjects were collected for the detection of the expression of ClqRp and gClqR by flow cytometry. The serum levels of Clq and ClqAb were detected by single radial immunodiffusion and enzyme-linked immunosorbent assay(ELISA)re- spectively. The correlation between ClqRp and gClqR and other disease activity parameters, such as Clq, ClqAb, SLEDAI score, anti-dsDNA antibody, C3, C4 levels were analyzed. Results The expression of ClqRp on mononuclear and neutrophiles of SLE was(7.2?2.3)% and(3.4?2.1)%, lower than that in healthy individuals [(10.6?2.1)% and(9.0?8.7)% ], P

Humans ; Lung Neoplasms ; therapy ; Medicine, Chinese Traditional ; methods ; trends

OBJECTIVE: To construct the expressing vector of siRNA which can inhibit the Smad3 activity. METHODS: Sixty-four bases of 2 pair oligos for hairp in RNA expression which targeted Smad3 gene were chemically synthesized and annealed. pSUPER vector was linearized with BgL II and Hin d III treated with alkaline phosphatase (CIP). Anneled oligos were inserted into the downstream of the treated pSUPER's pol III H1 promoter to construct RNAi plasmid (pSUPER Smad3). Oligos with a scrambled sequence were used as a negative control. pSUPER Smad3 was transfected into human renal tubular epithelial cells (HKC). RESULTS: Recombinant pSUPER Smad3 vector was identified by the digestion with Eco R I and Hin d III, and confirmed by the sequencing analysis with T3 primer. Sixty-four bases had been inserted into the expected site. Furthermore, the insertion sequence was exactly corrected. The activity evaluation indicated that mRNA and protein of Smad3 but not Smad2 were inhibited by pSUPER Smad3 in HKC. CONCLUSION The pSUPER Smad3 system has been constructed successfully, and has high inhibition and specificity in vitro.
Epithelial Cells ; metabolism ; Humans ; Kidney Tubules ; cytology ; Plasmids ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Smad3 Protein ; antagonists & inhibitors ; genetics ; Transfection

OBJECTIVETo study the change in ultrastructure of C6 glioma cells after photodynamic therapy (PDT), to compare morphological differences in necrosis and apoptosis before and after PDT treatment, and to evaluate the effect of photodynamic therapy on the blood brain tumor barrier (BTB) of C6 glioma.

METHODSThe model was produced by transplanting C6 glioma cells cultured in vitro using Peterson method into the caudate nuclei of Wister rats. The experiment group received PDT for two weeks after the operation. The sub-cellular structure, blood-brain-barrier (BBB) and BTB in both groups were observed under electron microscope.

RESULTSApoptosis in different phases and necrosis could be observed in some C6 glioma cells. Swelling occurred on the ultrastructure of cellular organs such as mitochondria and endoplasmic reticulum in most of the cells. Damage to the BTB, reduction of the number of cellular organs in endothelial cells of the capillary blood vessels, stretch of the tight junction, and enlargement of the gaps between endothelial cells were also seen in the experiment group. Meanwhile, limited impact on the normal sub-cellular structures and BBB was observed.

CONCLUSIONPDT could induce apoptosis and necrosis of C6 glioma cells due to the damage to the ultrastructure of mitochondria and endoplasmic reticulum. The weakened function of C6 glioma BTB initiated by PDT makes it possible to perform a combined therapy of PDT and chemotherapy for glioma.

Animals ; Blood-Brain Barrier ; Brain Neoplasms ; drug therapy ; ultrastructure ; Cell Line, Tumor ; Glioma ; drug therapy ; ultrastructure ; Photochemotherapy ; Rats

The purpose of this study was to investigate the effect of bisphosphonate combined with chemotherapy on bone mineral density (BMD) of patients with multiple myeloma (MM) and analyse its value of BMD detection in clinic of these patients. 53 MM cases were enrolled in this study, including 33 newly diagnosed, 10 refractory/relapsed and 10 stable cases. They were divided randomly into two groups, 33 MM cases were treated with bisphosphonates combined with chemotherapy and 20 MM cases were treated with chemotherapy alone. The chemotherapy schedules for all patients were same. BMD was tested using the dual energy X-ray absorptiometry at 2 time points, i.e. pretreatment (basal level) and 12 months after treatment with chemotherapy and bisphosphonates. Comparisons was performed with t tests using SPSS 11.0 software. The results indicated that there was minor difference between 2 groups for BMD scores of whole body and lumbar vertebra (L1-L4), but no difference for scores of the near-end of left femur. After treatment for 12 months, all BMD scores (whole body, lumbar vertebra and the near-end of left femur) increased significantly in the bisphosphonate combined with chemotherapy group (P < 0.05). In contrast, only minor changes were seen in chemotherapy alone group. It is concluded that the bisphosphonate combined with chemotherapy has displayed promotive effect on BMD of MM patients, the detection of BMD is sensitive and special method for monitoring therapeutic effect of bisphosphonate in MM patients, thus has useful value in clinic of these patients.
Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Bone Density ; drug effects ; Diphosphonates ; administration & dosage ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; metabolism ; pathology

OBJECTIVETo design and synthesize ribozymes targeting 138 and 218 sites of the mRNA nucleotide of mouse caspase-12, a key intermedium of ER stress mediated apoptosis, and to identify their activities through in vitro transcription and cleavage.

METHODSThe mouse caspase-12 gene fragment was obtained by RT-PCR and cloned into the PGEM-T vector under the control of T7 RNA polymerase promoter. The transcription product of the target was labeled with a-32P UTP, while ribozymes were not labeled. Ribozyme and target RNA were incubated for 90 min at 37 degree C in a reaction buffer to perform the cleavage reaction.

RESULTSIt was found that under a condition of 37 degree C, pH 7.5 and with Mg2+ in a concentration of 10 mmol/L, Rz138 and Rz218 both cleaved targets at predicted sites, and the cleavage efficiency of Rz138 was 100%.

CONCLUSIONRz138 and Rz218 prepared in vitro possess the perfect specific catalytic cleavage activity. Rz138 has excellent cleavage efficiency. It may be a promising tool to prevent ER stress induced apoptosis through catalytic cleavage of caspase-12 mRNA in vivo. It also can be used to verify whether caspase-12 is necessary in ER stress induced apoptosis.

Animals ; Base Sequence ; Caspase 12 ; genetics ; Endoplasmic Reticulum ; metabolism ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Oxidative Stress ; genetics ; RNA, Catalytic ; chemistry ; genetics ; RNA, Messenger ; genetics

OBJECTIVETo design hammerhead ribozymes against mouse caspase-7 and to study their expression and cleavage activity in vitro.

METHODSThe secondary structures of ribozyme and caspase-7 genes were analyzed and simulated by computer. Ribozymes DNA sequences were synthesized by automatic synthetic apparatus. Caspase-7 DNA sequence was acquired by reverse transcription PCR. Ribozymes and caspase-7 DNA sequences were separately cloned into pBSKneo U6 and pGEM-T vectors. Ribozymes and caspase-7 mRNA were obtained by transcription in vitro, and ribozymes cleavage activity was identified by cleavage experiment in vitro.

RESULTSTwo ribozymes named Rz333 and Rz394 targeting 333 and 394 sites in caspase-7 mRNA were designed by computer software, and their DNA sequences were synthesized. The expression vector of caspase-7 and plasmids containing Rz333 and Rz394 were reconstructed successfully. Ribozymes and caspase-7 mRNA were expressed by in vitro transcription. In vitro cleavage experiments showed that Rz333 cleaved caspase-7 mRNA and produced 243nt and 744nt segments. The cleavage efficiency is 67.98%, while Rz394 cannot cleave caspase-7 mRNA.

CONCLUSIONSRz333 can site-specifically cleave caspase-7 mRNA.

Animals ; Base Sequence ; Caspase 7 ; Caspase Inhibitors ; Caspases ; genetics ; Cloning, Molecular ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; RNA, Catalytic ; biosynthesis ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics

Computer Graphics ; Computer-Assisted Instruction ; Forensic Pathology ; Humans ; Imaging, Three-Dimensional ; Pathology ; education ; Pathology, Clinical ; Specimen Handling ; methods

The first hand documents, the process and achievements of Mr. CHENG' s study in Japan in the 1930s were investigated in order to provide basic information to explore the original academic thoughts of Mr. CHENG and the Chengjiang Acupuncture School. It was concluded that the experiences of Mr. CHENG's study in Japan had significant influence on enriching and developing his academic thoughts. It has not only enhanced his confidence in therapeutic effect of moxibustion and revival of Chinese acupuncture, but also established the foundation for the reform of needles and expansion of the domain of Chinese acupuncture and its development after returning home.
Acupuncture ; education ; history ; Acupuncture Therapy ; history ; China ; History, 19th Century ; History, 20th Century ; Humans ; Japan ; Male

OBJECTIVETo investigate the secondary operation methods and the effects on the prognosis of unexpected gallbladder cancer (UGC).

METHODSA retrospective analysis on the clinical data was made for 41 patients who underwent extended radical excision from June 1995 to December 2002. Among the patients, 12 were male, 29 were female. The average age was 51 years old. The 41 patients had undergone gallbladder excision because of cholecystitis complicated lithiasis of gallbladder (32 cases), polypi of gallbladder or adenoma (9 cases). Postoperative pathology showed that 32 cases were adenocarcinoma of gallbladder, 6 cases were squamous carcinoma, 3 cases were squamous adenocarcinoma. Six cases were on the stage of Nevin I, 16 on Nevin II, 17 on Nevin III, 2 on Nevin IV. The second operation was performed after 6-30 d of the first operation. The second operation chose the improved method of Glenn excision of carcinoma of gallbladder.

RESULTSOn the second operation, 14 cases were with lymphatic metastasis, 14 with gallbladder metastasis, 6 with bile duct metastasis, 2 with pancreas metastasis. Fourteen cases were on the stage of Nevin IV, 9 on Nevin V, none on Nevin I, II and III. After the second operation, 1 year survival rate was 100% (41 cases); The three-year survival rate was 53.8% (22 cases); The five-year survival rate was 17.5% (7 cases).

CONCLUSIONExtended radical excision is one of the most important methods for the treatment of UGC.

Adult ; Aged ; Cholecystectomy ; methods ; Diagnostic Errors ; Female ; Gallbladder Neoplasms ; diagnosis ; mortality ; surgery ; Humans ; Male ; Middle Aged ; Reoperation ; Retrospective Studies ; Survival Rate