Objective To establish tetracycline-controlled inducible system(Tet-on) in HepG2 cell for further research of the function of related gens.Methods The HepG2 cells were transfected with pWHE146 vector by using liposome transfection reagent.The transfected cells were selected in medium containing G418,and G418-resistant clones were isolated.All individual G418-resistant clones were screened by transient transfection with plasmid pTRE-hyg-luc for clones with low background and high induction of luciferase in response to Dox.Results One HepG2 cell line,which exhibited high levels of induction(154,106 times)and high gene expression levels,was obtained.Conclusions The HepG2 cell line can be used to highly express eukaryotic gene and this Tet-on system is available for use in eukaryotic gene function studies.

Secondary hyperparathyroidism (SHPT) is a common complication in patients with chronic renal failure (CRF).Medical treatment may be the first choice for most SHPT patients,but parathyroidectomy (PTX) is neededwhen medical treatment doesn't work in patients with refractory SHPT.However,there is no strong evidence to support which surgical procedure is the most effective one for patients with SHPT.An interdisciplinary discussion between nephrologists and surgeons is needed when choosing a preferred surgical method for refractory SHPT patients.The factors must be discussed including:the patient's age,the underlying kidney disease,clinical symptoms,the level of intact parathyroid hormones (iPTH) and blood calcium,the patient's ability to obtain and comply with medication treatment and the estimated duration of dialysis before kidney transplantation.Refractory SHPT patients will benefit from the surgical operation treatment and reoperation of recurrent SHPT is still an ideal treatment method.

70 patients with gastric carcinoma were studied by EUS prior to surgery. The results were correlated with the histology of resected specimens according to the new TNM classification. EUS was accurate in assessing the depth of tumor infiltration, the overall accuracy of EUS was 74.3%. The cancerous ulcer and obstruction are the main causes of over- and understaging, respectively. EUS was relatively accurate in the assessment of lymph node metastasis, the overall accuracy was 57.1%. However, negative-predictive rate is lower, about 42.9%. It is difficult to distinguish between inflammatory and metastatic lymph nodes. EUS was not reliable in diagnosing distant metastasis, due to its limited depth of penetration. In our experience, in staging the gastric carcinoma, greater accuracy would be achieved if we use EUS for T and N factors, and CT for M factor.

Objective To investigate the value of endoscopic ultrasonography ( EUS) in diagnosing pancreatic pseudocyst. Methods EUS was performed in 35 cases with pancreatic pseudocyst. Results Of the series, 41 cysts found in the head, body, tail and both body and tail of the pancreas were 13 , 3 , 19 and 6 respectively. The cystic wall was smooth in 29 cysts and rough in 6; good signal in cystic fluid 19, floccu-lar echo 16, cystic septum 1 and calcification of cyst wall 1, associated ductus pancreaticus expansion in 7, pancreas parenchyma uneven echo in 21, pancreas parenchyma calcification in 4, pancreas atrophy in 2, pancreas carcinoma in 2 , and normal pancreas in 6, protuberances impression on gastrointestinal tract 5 ( duodenal descending part obstruction, n = 1 ) stomach varicose vein in fundus 4 and digestive tract hemorrhage 2. Conclusion EUS may clearly show cyst's size, position, configuration and relation with the pancreas, also EUS guided FNA can be performed, so it has significance in diagnosing and distinguishing pancreatic pseudocyst.

Objective By contrasting and analyzing the clinic manifestations, radiological features and pathological results of patients with bone invasion from skin of the lower limbs that were associated with malignancy, authors advanced X-ray diagnostic evidence to promote its diagnostic accuracy. Methods The radiological appearances of squamous cell carcinoma arising in chronic ulcers, sinus and scars of the lower limbs in 16 patients from February 1968 to April 2002 confirmed by pathology, were analyzed retrospectively. All cases were taken posterior-anterior and lateral radiograph before operation. There were 15 males and 1 females aging from 15 to 67 years (mean, 50.4 years). The lesion was located at the front of tibia in 15, and the middle inferior part of femur. The medical history of primary diseases varied from 1 to over 50 years with an average of 20 years and 3 months. Of 16 cases, the duration of 8 cases from a sudden change of state of an illness to the final diagnosis was 4 to 6 months, 4 cases above 1 year, and 4 cases were found with no any change of state of an illness. Results There were 14 cases observed as typical chronic osteomyelitis, which demonstrated peripheral osteolytic defect, 13 cases with osteolytic destruction spreading outwards from front of tibia, and 1 case with widespread osteolytic destruction in the middle and inferior part of femur. Floating-ice-like rudimental bone in the defect was found in 11 cases, and 4 case were observed with old fracture. 2 cases without obvious history of typical chronic osteomyelitis, 1 case with soft tissue trauma and 1 case of burn, showed small Saucer-like erosion of the cortex and huge lobar soft tissue masses. Moreover, patchy shadow and Codman's triangle were found in 1 case of burn. Furthermore, lobar soft tissue masses and huge ulcer accord with osteolytic destruction was observed in all cases, and soft tissue masses were larger than the area of bony destruction in 6 cases. Conclusion Roentgenogram could display shape, location and extension of bone invasion from skin of the lower limbs that were associated with malignancy, which contribute to diagnose the lesion and provide useful evidence for surgical plan.

Objective To study the effects of HIF-1α expression on the growth of subcutaneously implanted human hepatocellular carcinoma (HCC) cell lines in nude mice. Methods A nude mouse model of subcutaneously implanted HCC cell line HepG2Tet-on-HIF-1α was established. This cell lines characterizes the inducible expression of HIF-1α by doxycycline (Dos). The impact of HIF-1α expression induced by Dox on the growth of subcutaneously implanted HCC cell lines in the nude mouse model was observed. Results The mRNA and protein expression of HIF-1α were significantly up-regulated in the nude mouse model by oral administration of Dox. Compared with that model in which Dox was not administrated ie Dos( - ) group, the tumor volume(513.545 ± 276. 229) mm3 vs. ( 166. 506 ± 110. 142) mm3 ( P < 0. 05 ), tumor weight ( 1.251 ± 0. 438 ) g vs. (0. 640 ± 0. 296) g ( P < 0. 05 ), and tumor growth velocity were significantly enhanced in Dox ( + ) group, while tumor necrosis was inhibited ( 31. 360% ±2. 728% vs. 36. 640% ± 3. 804% ) (P<0. 05). The weight loss of nude mice was larger in Dox( + )group( P < 0. 01 ). There was no liver or lung metastasis in either group. Conclusion The expression of HIF-1αin subcutaneously implanted HCC in a nude mouse model is up-regulated by oral Dox. High grade expression of HIF-1α promotes the growth of implanted HCC.

Objective To investigate the effects of different doses of 1 ,25(OH)2D3 on rat Thy‐1 nephritis mesangial cell ap‐optosis .Methods 120 clean SD rats were divided into the blank control group(group A) and the experimental group .The experi‐mental group was re‐divided into the nephritis group(group B) ,0 .25μg 1 ,25(OH)2D3 intervention group(group C) and 0 .50μg 1 , 25(OH)2D3 group(group D) ,30 cases in each group .The group C and D were performed the medication intervention after success‐fully constructing the model .6 rats were randomly killed in each group on 1 ,3 ,7 ,14 ,21 d after intervention .The renal tissues were taken for determining the renal pathological injury classification after hematoxylin and eosin staining and PAS staining .The expres‐sion of Caspase‐3 in the renal tissues was detected by the immunohistochemistry method ,and the glomerular cell apoptosis was de‐tected by TUNEL .Results The immunohistochemistry results showed that compared with the group A ,the Caspase‐3 expression in the group B was increased (P<0 .05);compared with the group B ,the Caspase‐3 expression in the group C and D was increased , but there was no statistical differences between them (P> 0 .05);the Caspase‐3 expression in the group D was highest ,but the difference was not statistically significant compared with the group C .The TUNEL results showed that compared with the group A ,the apoptotic glomerular cells in the group B were increase obviously (P<0 .05) ,which was gradually enhanced over time ;com‐pared with the group B ,the apoptotic glomerular cells in the group C and D were increased significantly (P<0 .05) ,reaching a peak on 3 d ,then decreasing gradually on 7 ,14 d and tending to be normal on 21 d(P<0 .05);the comparison between the group C and D showed that the apoptotic cells in the group D were increased significantly ,but the difference between these two group ,was not statistically significant(P>0 .05) .Conclusion 1 ,25(OH)2D3 has the effect for inducing glomerular mesangial cell apoptosis in rat , which participates in the regulation of Caspase‐3 ,induces glomerular mesangial cell apoptosis ,promotes the recovery of nephritis and could delay the progression of renal disease .

Objective To investigate the clinical efficacy of a new-type moxibustion box in treating cervical spondylotic radiculopathy.Methods Sixty patients with cervical spondylotic radiculopathy were randomly allocated to treatment and control groups, 30 cases each. The treatment group received nape-eight-needle acupuncture plus new-type moxibustion box therapy and the control group, nape-eight-needle acupuncture plus conventional moxibustion box therapy. After 10 treatments, the NPQ score, the symptom and sign score and the VAS score were recorded in the two groups and the clinical therapeutic effects were compared between the two groups.Results The total effective rate was 100.0% in the treatment group versus 83.3% in the control group, and the difference was statistically significant (P<0.05). There were statistically significant pre-/post-treatment differences in the NPQ score, the symptom and sign score and the VAS score in the two groups (P<0.01,P<0.05). There were statistically significant post-treatment differences in the NPQ score, the symptom and sign score and the VAS score between the treatment and control groups (P<0.05).ConclusionNape-eight-needle acupuncture plus a new-type moxibustion box is an effective way to treat cervical spondylotic radiculopathy.

Objectives:To investigate the significance of diagnosis and treatment guided by EUS in locating lesion of pancreas. Methods: Transgastric aspiration biopsy and placeing plastic stent were done by endoscopic ultrasonography, Olympus GF UM30P with 18G needle. Results: The successful rate of puncture and accuracy of diagnosis are both 100%.The diameters of all of the pancreatic cysts were less than 50% of that after the operation,two pancreatic cysts disappeared.Transgastiric placeing plastic stent was done in one patient,and its cyst was less than 50% after 1 week. Five pancreatic cysts were less than 50% after three six.There is no early and delay complications. Conclusions: Diagnosis and treatment guided by EUS in location lesion of pancreas is useful.