The pol gene of HIV-1 encodes mainly three enzymes: reverse transcriptase (RT), protease (PR) and integrase (IN). Currently, FDA approved drugs targeting RT and PR are available and administered in various combinations, while no anti-IN drug was approved. HIV-1 integrase is an essential enzyme for the viral replication and an interesting target for the design of new pharmaceuticals for multi-drug therapy of AIDS. The 288 amino acids of IN (32kDa) recognizes specific sequences in the long terminal repeats (LTRs) of the retroviral DNA. The IN protein catalyzes the 3′-processing step and the 5′-strand transfer step reaction in vivo, which was called integration and this reaction could be analysed by ELISA Assay in vitro. It has been reported that F185K and C280S mutations of HIV-1 integrase would improve the enzyme solubility, and the catalytical activity of the enzyme was the same as that of the wild-type enzyme in vitro. In order to build the platform of screening inhibitor against integrase of HIV-1 virus, the IN enzyme was expressed and the function of integrase protein was assayed. The cDNA of clade B HIV-1 genome was used as a template, overlapped PCR was used to construct site mutagenesis of F185K/C280S and NdeI/Xho I restriction sites were brought in. The PCR product was cloned into the prokaryotic vector pET-28a(+) to form a recombined plasmid, transferred into the host cell E.coli(BL21 DE3). The recombined clones were identified by PCR and Nde I/Xho I digestion .The positive plasmid was sequenced, and the successfully recombined plasmid in the host cell was induced by IPTG. The expressed IN protein was puriied sy the Co+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombined IN protein. The recombinant protein was soluble, and expressed highly and stably in E.coli. The molecular weight of the expression product was identical to the expectation. The IN protein was proved to be functional in 3′ processing and 5′strand transfer by ELISA. It will be helpful to build the platform of screening inhibitors against HIV-1 integrase.

OBJECTIVETo investigate the potential of gene therapy of rat prolactinomas mediated by adenoviral vectors with a gene encoding rat tyrosine hydroxylase.

METHODSRecombinant replication-deficient adenovirus named Ad-GFP-TH with rat TH-cDNA and control adenovirus named Ad-GFP were constructed by homologous recombination in bacterial cells. The rat pituitary prolactinoma cell line MMQ are chosen as the target cells to study the effect of gene therapy on their growth and prolactin secretion mediated by Ad-GFP-TH.

RESULTSRecombinant Ad-GFP-TH and Ad-GFP were successfully reconstructed. Transfection of MMQ cells with Ad-GFP-TH not only restrained their growth but also decreased their PRL secretion.

CONCLUSIONGene therapy may serve for a potential treatment for prolactinomas, especially invasive prolactinomas.

Adenoviridae ; genetics ; Animals ; Genetic Therapy ; Genetic Vectors ; Pituitary Neoplasms ; therapy ; Prolactinoma ; therapy ; Rats ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Tyrosine 3-Monooxygenase ; biosynthesis ; genetics

OBJECTIVETo evaluate clinical outcome after anterior cruciate ligament (ACL) reconstruction with double-bundle or single-bundle by meta-analysis.

METHODSRandomized controlled trials (RCTs) on differences of clinical outcomes of ACL reconstruction were retrieved in Ovid Medline, PubMed, Embase, Cochrane Library, CBM and VIP database. Relevant journals or conference proceedings were also searched manually. Then extracted the date of KT-1000 arthrometer, pivot-shift testing, Lysholm score and IKDC final score in these researches. RevMan 5.0.23 software was used for data analyses.

RESULTSEight prospective RCTs met the inclusion criteria. The combined results of meta-analysis indicated that there was statistical difference between two operative procedures on postoperative KT-1000 arthrometer side-to-side [WMD = -0.35, 95%CI (-0.61, -0.08), P = 0.01], Lysholm score [WMD = -1.91, 95%CI (-3.45, -0.37), P = 0.01]. But the difference of KT-1000 arthrometer side-to-side is demonstrated to be clinically insignificant. Others indicated that there was no statistical differences with respect to IKDC final score [OR = 1.80, 95%CI (0.98, 3.31), P = 0.06], having a normal or nearly normal pivot-shift testing [OR = 1.64, 95%CI (0.85, 3.16), P = 0.14].

CONCLUSIONSDouble-bundle reconstruction does not result in clinically significant advantage when compared with single-bundle. The results do not support the theory that double-bundle reconstruction controls knee rotation better.

Anterior Cruciate Ligament ; surgery ; Humans ; Joint Instability ; surgery ; Reconstructive Surgical Procedures ; methods ; Treatment Outcome

The purpose of this study is to summarize teaching experiences on channel and acupoints. The method of classified teaching is put forward and approached to explore the law of acupoint teaching. It is held that the method could reduce randomness and enhance appropriateness, rationality and scientific validity of the teaching. Class hours are thus optimized and the best teaching result achieved. Acupoints are classified into 4 categories of core points, focal points, regular points and ordinary points according to their use frequency and theoretical importance. Classification methods, principles and categories are then determined. And content of courses, teaching techniques and requirements are introduced as well. Suggestions are offered on enriching teaching contents, for instance, paraphrasing of the acupoint names, indications, point prescriptions and clinical experiences. Thus the conclusion is made that it is imperative to reform the conventional sort of acupoints classified by only focal points and ordinary points as well as the related teaching methods.
Acupuncture ; education ; standards ; Acupuncture Points ; Cyclobutanes ; Humans ; Teaching ; standards

Objective To explore the effect and indication of splenectomy in liver transplantation.Methods From January 2001 to April 2006,260 patients underwent piggyback orthotopic liver transplantation(PBOLT),and 28 patients had undergone combined PBOLT and splenectomy(splenectomy group).These patients were compared to 56 randomly selected non-splenectomy patients from the same transplant period,meaningly two controls were se- lected for every non-spleneetomy case.Two groups were analyzed with respect to rate of infection and survival rate, as well as biopsy-proven acute allograft rejection within 30 days after transplantation.Results Rate of infection in the splenectomy group was higher than that in the non-splenectomy patients(85.7% vs 55.4%,P

That eleven patients with sellar germinomas were confirmed by pathological examinations were retrospectively reviewed. In the 11 patients, 2 were males and 9 were females. Mean age was 16.8 years old and mean history was 2.6 years. Among them, 11 patients had diabetes insipidus, 6 patients had visual disturbances, 5 patients had retarded growth, and 5 patients had hypopituitary function. The diameter of tumor was 1-3 cm in 8 cases and greater than 3 cm in 3 cases. MRI showed hypointensity or homogeneous on T1-weighted images in 8 cases. The tumors were fibrous, purple grey or reddish in color with rich blood supply. Germinomas were confirmed by postoperative pathological examinations. All patients received postoperative irradiation. Postoperative follow-up was from 3 months to 6 years. Diabetes insipidus was improved in 9 cases. Visual function were improved in 4 cases, but hypopituitarism and retarded growth were unchanged. In conclusion, when MRI shows sellar parenchyma tumor with unclear boundary in young patients with diabetes insipidus and hypopituitarism, sellar germinoma should be highly suspected. Otherwise, radiation therapy may profitably aid to confirm the diagnosis since irradiation is the main treatment of sellar germinomas.
Adolescent ; Adult ; Brain Neoplasms ; diagnosis ; surgery ; Diagnosis, Differential ; Female ; Germinoma ; diagnosis ; surgery ; Humans ; Magnetic Resonance Imaging ; Male ; Pineal Gland ; Pinealoma ; diagnosis ; surgery ; Pituitary Neoplasms ; diagnosis ; surgery ; Retrospective Studies ; Sella Turcica

OBJECTIVETo analyze the relationship between invasive pituitary adenomas and cavernus sinus and cariod artery and to predict their surgical outcomes.

METHODSTotally 270 patients with pituitary tumors were investigated in this retrospective study, including 113 men and 157 women, with a mean age of 40.8 years. The mean disease history was 3.6 years. Pituitary microadenomas were diagnosed in 56 cases, macroadenomas in 118 cases, and hugeadenomas in 96 cases. Adrenocorticotropic hormone-releasing adenomas (Cushing's diseases) were diagnosed in 40 cases, growth hormone-releasing adenomas in 58 cases, prolactinomas in 57 cases, and non-functional pituitary adenomas in 115 cases. Transsphenoidal microsurgery were performed on 260 patients, while transcranial microsurgery on 6 patients.

RESULTSThe percentage of invasive pituitary adenomas was about 3.6% in microadenomas, 20.4% in macroadenomas, and 61.4% in hugeadenomas. Rate of total removal was 94.1% in non-invasive pituitary adenomas, and was 58.8% in invasive pituitary adenomas.

CONCLUSIONSIt is important to analyze the grade of invasive pituitary adenomas to improve the removal of pituitary adenomas, avoid injuring cariod artery, and increase the rate of total removal.

Adenoma ; pathology ; surgery ; Adolescent ; Adult ; Aged ; Carotid Arteries ; pathology ; surgery ; Cavernous Sinus ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Hypophysectomy ; methods ; Magnetic Resonance Imaging ; Male ; Microsurgery ; Middle Aged ; Neoplasm Invasiveness ; Pituitary ACTH Hypersecretion ; pathology ; surgery ; Pituitary Neoplasms ; pathology ; surgery ; Radiotherapy, Adjuvant ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo analyze the transsphenoidal microsurgical results of non-invasive prolactinomas, in order to provide reference for their treatment choice.

METHODSTo review the transsphenoidal microsurgical results of 234 non-invasive prolactinomas treated in our department in recent 10 years, and to analyze the prognostic factors. There were 18 males and 216 females, aged between 13 and 58 years, averaged (31.1 +/- 8.5) years. The course ranged from half a month to 20 years, averaged (47.3 +/- 44.9) months. The preoperative serum PRL level ranged between 41 and 8406 ng/ml, averaged (400.5 +/- 888.0) ng/ml, with a median of 164.1 ng/ml. The primary symptoms were amenorrhea, galactorrhea and/or infertility in 211 cases. The tumor size was small (< 1 cm) in 100, large (> or = 1 cm) in 116 and giant (> or = 3 cm) in 18 cases. All the patients received transsphenoidal microsurgery and were followed-up for 12 to 132 months, averaged (43.8 +/- 35.0) months.

RESULTSThere was no mortality. One hundred and twenty-seven (54.3%) cases had transient postoperative imbalance of water and electrolytes. One hundred and eighty-eight cases (80.3%) were cured, 12 (5.1%) experienced clinical remission, 20 (8.5%) were improved, and 14 (6.0%) were ineffective. The male patient, the giant prolactinomas and those with higher preoperative serum PRL level had a relative poor postoperative prognosis. While the other factors had no influence on prognosis, including the course, preoperative bromocriptine intake, tumor texture, tumor apoplexy and intraoperative descending extent of the diaphragm of sella. The overall operative expense for transsphenoidal microsurgery ranged from 8323.8 to 22898.5 yuan, averaged (12912.0 +/- 2361.2) yuan.

CONCLUSIONSTranssphenoidal microsurgery may be chosen as the primary therapy for non-invasive prolactinomas, with the purposes of therapeutical efficacy, facilitating the patients, re-establishing the patients' self-confidence and reducing the overall expense.

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Hypophysectomy ; methods ; Male ; Microsurgery ; Middle Aged ; Pituitary Neoplasms ; surgery ; Prolactinoma ; surgery ; Retrospective Studies ; Sphenoid Sinus ; surgery ; Treatment Outcome

OBJECTIVETo study the clinical characteristics, diagnosis and surgical effects of thyroid-stimulating hormone pituitary adenomas (TSH-omas).

METHODSThe clinical data of 19 patients (14 female and 5 male) with TSH-omas were analyzed retrospectively in this study from January 2001 to December 2008. The patients ranged from 20 to 70 years old (average 40.5 years old) and had disease histories from 1 to 228 months (average 55 months). Among these patients, 15 of them complained of thyrotoxicosis symptoms, while the other 4 patients' symptoms were associated with headache and/or visual disturbance caused by the tumor mass effect. Initially, 12 of the 15 patients with thyrotoxicosis symptoms were misdiagnosed with Grave's disease. As a result 2 of them received (131) Iodine, and one received subtotal thyroidectomy. All of these patients underwent transsphenoidal microsurgery.

RESULTSAverage follow-up period was 3.6 years (6 months-7 years). Pathological analysis of the surgical specimen showed pituitary adenoma in all patients, immunohistochemistry were positive for TSH in 17 cases, negative for TSH in 2, positive for growth hormone in 2, positive for prolactin in 1, and positive for adrenocorticotrophic hormone in 1. Postoperative MRI revealed that the tumors in 15 patients were removed totally, though 4 patients still had residual tumors. The thyroid hormone level tests suggested that 13 patients could be considered normal 3 months after their tumors were removed, though 2 of patients with normal postoperative MRI and thyroid hormones showed increased levels of TSH. For these 2 patients, tumors did not recur and their thyroid hormone levels returned to normal after pituitary radiotherapy. The cure rate was 11/19 after surgery and 13/19 after surgery plus pituitary radiotherapy.

CONCLUSIONSThe screening test for hyperthyroidism patients with high TSH levels is a key point to improve the accuracy rate in early diagnoses of TSH-omas. The transsphenoidal microsurgery is first choice to treat TSH-omas, while pituitary radiotherapy and somatostatin analogs are beneficially adjunctive therapies.

Adult ; Aged ; Female ; Humans ; Hyperthyroidism ; metabolism ; Male ; Middle Aged ; Pituitary Neoplasms ; diagnosis ; metabolism ; surgery ; Retrospective Studies ; Thyrotropin ; metabolism ; Young Adult

BACKGROUNDRecurrent ischemic events occurred even during routine use of 75 mg clopidogrel in addition to aspirin, that indicated a potentially insufficient maintenance dosage of clopidogrel. The aim of the present study was to evaluate the short-term efficacy and safety of a 150 mg maintenance dose of clopidogrel following a 600 mg loading dose in patients with an acute coronary syndrome (ACS) undergoing drug eluting stent (DES) implantation.

METHODSBetween November 2005 and November 2006, a total of 813 consecutive ACS patients undergoing DES implantation were enrolled. A 600 mg loading dose was administered before percutaneous coronary intervention (PCI) and patients were randomized to receive clopidogrel 75 mg or 150 mg for 30 days in addition to 300 mg aspirin daily. Primary end points were the composite of cardiac death, non-fatal myocardial infarction (MI) and urgent target vessel revascularization (UTVR). Secondary end points included stent thrombosis (ST), major and minor bleeding events at 30 days.

RESULTSAt a follow-up period of 30 days, 4 (1.0%) patients in the 150 mg group and 9 (2.2%) patients in the 75 mg group (P > 0.05) reached the primary end points. There was no significant difference in the incidences of MI (0.5% vs 1.2%, P > 0.05), UTVR (0.7% vs 2.0%, P > 0.05), and cardiac death (0.2% vs 0.2%, P > 0.05) between the two groups. The incidence of ST (0 vs 1.5%, P < 0.05) was significantly lower in the 150 mg group than that in the 75 mg group. There were no significant differences between both groups regarding the risk of major (0.2% vs 0, P > 0.05) or minor (0.5% vs 0.2%, P > 0.05) bleedings.

CONCLUSIONA high clopidogrel maintenance dose of 150 mg daily following a 600 mg loading dose for the first month after PCI procedure reduces the risk of ST and appears to be safe in patients with ACS undergoing DES implantation.

Acute Coronary Syndrome ; therapy ; Aged ; Angioplasty, Balloon, Coronary ; methods ; Coronary Angiography ; Drug-Eluting Stents ; Female ; Humans ; Male ; Middle Aged ; Platelet Aggregation Inhibitors ; administration & dosage ; therapeutic use ; Ticlopidine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Treatment Outcome