Objective To approach the surgical timing for the conjoined twins, location of the separation line for the joined liver, and the separation method.Methods The common bile ducts of the conjoined twins were considered as two vertical lines, and a vertical line running parallel to the two lines was set as the separation line for the joined liver.Local blood flow blocking method was then used to separate the joined liver.Results Among all the three cases of the conjoined twins, one case was with sternoxiphopagus and the other two with thoracoabdominalpagus.All the three cases of con-joined twins shared the common livers, but each case had respectively separated gallbladders and bile ducts.They underwent the surgical separation at the age of 28 d and 96 d and 89 d successfully.Their liver sections bled rarely by blocking the local blood flow.The liver function recovered successfully af-ter the operation.All the 6 sick children recovered and were discharged from our hospital.Conclusion Porvided the conjoined twins shared the joined liver with respectively separated common bile ducts, in most cases, the injuries of the important liver vascular as well as bile ducts could be avoided when the separation line for the joined liver was selected with the common bile ducts of the conjoined twins as the longitudinal coordinate.The local blood flow blocking method only blocked the local blood flow, in-terfering to the liver blood flow in the non-operating areas rarely, which was instrumental in the recovery of the liver function and increase of the survival rate of the conjoined twins after the operation.

Objective To evaluate the safety and efficacy of transumbilical single port laparoscopic cholecystectomy. Methods The clinical data of 16 patients who received transumbilical single port laparoscopic cholecystectomy at Xinqiao Hospital from January 2008 to May 2010 were retrospectively analysed. An incision with a length of 1.5 cm was made adjacent to the umbilicus, and then two 5 mm trocars and one 10 mm trocar were installed. After the establishment of pneumoperitoneum, a laparoscopic camera was placed via the 10 mm trocar,and laparoscopic instruments and a 5 mm ultrasonic scalpel were placed via the two 5 mm trocars, respectively.Cholecystectomy was performed in the same manner as for the conventional laparoscopic procedure. Results All the operations were successfully carried out. The operation time was 50-150 minutes. No drainage tube was inserted,and no complications such as bleeding or bile leakage were observed after the operation. Patients recovered well,and no scarring was observed around the umbilicus. Conclusions Transumbilical single-port laparoscopic cholecystectomy is safe and feasible, but it is more difficult than laparoscopic cholecystectomy in terms of manipulation.Transumbilical single-port laparoscopic cholecystectomy has the potential to replace laparoscopic cholecystectomy if the operative instruments are improved.

Algorithms ; Carrier Proteins ; chemistry ; Crystallography, X-Ray ; Humans ; Magnetic Resonance Spectroscopy ; Molecular Dynamics Simulation ; Nuclear Proteins ; chemistry ; Principal Component Analysis ; Protein Structure, Tertiary ; Proteins ; chemistry ; Scattering, Small Angle ; X-Ray Diffraction ; methods

Uch37 is a de-ubiquitinating enzyme that is activated by Rpn13 and involved in the proteasomal degradation of proteins. The full-length Uch37 was shown to exhibit low iso-peptidase activity and is thought to be auto-inhibited. Structural comparisons revealed that within a homo-dimer of Uch37, each of the catalytic domains was blocking the other's ubiquitin (Ub)-binding site. This blockage likely prevented Ub from entering the active site of Uch37 and might form the basis of auto-inhibition. To understand the mode of auto-inhibition clearly and shed light on the activation mechanism of Uch37 by Rpn13, we investigated the Uch37-Rpn13 complex using a combination of mutagenesis, biochemical, NMR, and small-angle X-ray scattering (SAXS) techniques. Our results also proved that Uch37 oligomerized in solution and had very low activity against the fluorogenic substrate ubiquitin-7-amino-4-methylcoumarin (Ub-AMC) of de-ubiquitinating enzymes. Uch37Δ(Hb,Hc,KEKE), a truncation removal of the C-terminal extension region (residues 256-329) converted oligomeric Uch37 into a monomeric form that exhibited iso-peptidase activity comparable to that of a truncation-containing the Uch37 catalytic domain only. We also demonstrated that Rpn13C (Rpn13 residues 270-407) could disrupt the oligomerization of Uch37 by sequestering Uch37 and forming a Uch37-Rpn13 complex. Uch37 was activated in such a complex, exhibiting 12-fold-higher activity than Uch37 alone. Time-resolved SAXS (TR-SAXS) and FRET experiments supported the proposed mode of auto-inhibition and the activation mechanism of Uch37 by Rpn13. Rpn13 activated Uch37 by forming a 1:1 stoichiometric complex in which the active site of Uch37 was accessible to Ub.
Binding Sites ; Catalytic Domain ; Chromatography, Gel ; Crystallography, X-Ray ; Humans ; Membrane Glycoproteins ; chemistry ; genetics ; metabolism ; Nuclear Magnetic Resonance, Biomolecular ; Protein Binding ; Protein Conformation ; Protein Multimerization ; Scattering, Small Angle ; Ubiquitin Thiolesterase ; chemistry ; genetics ; metabolism ; Ultracentrifugation

Papaya, which is mainly cultivated in the southeastern region of China, is one of the four famous fruits in Lingnan. It is favored by people because of its edible and medicinal value. Fructose-6-phosphate, 2-kinase/fructose-2, 6-bisphosphatase (F2KP) is a unique bifunctional enzyme with a kinase domain and an esterase domain that catalyzes the synthesis and degradation of fructose-2, 6-bisphosphate (Fru-2, 6-P₂), an important regulator of glucose metabolism in organisms. In order to study the function of the gene CpF2KP encoding the enzyme in papaya, it is particularly important to obtain the target protein. In this study, the coding sequence (CDS) of CpF2KP, with a full-length of 2 274 bp, was got from the papaya genome. The amplified sequence of full-length CDS was cloned into the vector PGEX-4T-1 which was double digested with EcoR I and BamH I. The amplified sequence was constructed into a prokaryotic expression vector by genetic recombination. After exploring the induction conditions, the results of SDS-PAGE showed that the size of the recombinant GST-CpF2KP protein was about 110 kDa. The optimum IPTG concentration and temperature for CpF2KP induction were 0.5 mmol/L and 28 ℃, respectively. The purified sin[A1] gle target protein was obtained after purifying the induced CpF2KP protein. In addition, the expression level of this gene was detected in different tissues, and showed that the gene was expressed at the highest level in seeds and the lowest in pulp. This study provides an important basis for further revealing the function of CpF2KP protein and studying the involved biological processes of this gene in papaya.
Humans ; Carica/genetics* ; Recombinant Proteins ; Carbohydrate Metabolism ; Cloning, Molecular ; China