Drugs, Chinese Herbal ; Humans ; Integrative Medicine ; Medicine, Chinese Traditional

In 1989, Fitz defined and classified severe acute pancreatitis (SAP) and he thought that surgical treatment of SAP is feasible. Since then, the strategy of treating SAP was a controversial issue over past decades. Currently, relevant literatures reported that medical or minimally invasive treatments are superior to surgical treatment, while it is too early to make the final conclusion because of the complexity of SAP. From 1989 to 2008, 1852 patients with acute pancreatitis were treated at the General Hospital of PLA, and the clinical data of 18 patients who died of SAP were retrospectively analyzed. Based on the analysis, we concluded that medical conservative therapy and surgical operative therapy should not be opposed to each other. Selecting ideal timing and appropriate operation on the basis of pathological changes of retroperitoneum and pancreatitis should be considered seriously, and the new concept of "miniaturization of trauma" should also be introduced in treating SAP.

In order to construct and express human cardiac troponin C-linker-troponin I(P) [ cTnC-linker-TnI(P)] fusion protein, detect its activity and prepare lyophilized protein, we searched the CDs of human cTnC and cTnI from GenBank, synthesized cTnC and cTnI(30-110aa) into cloning vector by a short DNA sequence coding for 15 neutral amino acid residues. pCold I-cTnC-linker-TnI(P) was constructed and transformed into E. coli BL21(DE3). Then, cTnC-linker-TnI(P) fusion protein was induced by isopropyl-β-D-thiogalactopyranoside (IPTG). Soluable expression of cTnC-linker-TnI(P) in prokaryotic system was successfully obtained. The fusion protein was purified by Ni²⁺ Sepharose 6 Fast Flow affinity chromatography with over 95% purity and prepared into lyophilized protein. The activity of purified cTnC-linker-TnI(P) and its lyophilized protein were detected by Wondfo Finecare™ cTnI Test. Lyophilized protein of cTnC-linker-TnI(P) was stable for 10 or more days at 37 °C and 4 or more months at 25 °C and 4 °C. The expression system established in this research is feasible and efficient. Lyophilized protein is stable enough to be provided as biological raw materials for further research.
Escherichia coli ; Freeze Drying ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; Troponin C ; biosynthesis ; Troponin I ; biosynthesis

A 37-year-old male patient presented with persistent generalized itching erythema and papules for more than 1 month.The patient had received surgical treatment for type B3 thymoma in stage Ⅳ.Five months prior to the presentation,he developed myasthenia gravis.He also complained of chronic diarrhea for two years.Physical examination revealed white patches in the oral mucosa as well as scaly erythema of varying size on the face,trunk and extremities.Lamellar scales could be seen after scratching,while removal of scales could not result in the appearance of bleeding points.There was scaly hyperkeratotic erythema of palms and soles.Histopathological examination revealed psoriasis-like epidermal hyperplasia and parakeratosis with multiple dyskeratocytes,and some of the dyskeratocytes were surrounded by lymphocytes.There was a perivascular infiltration with a small number of lymphocytes.Immunohistochemical study showed positive staining for CD3,CD4 (dermis),CD8 (epidermis) and CD45RO,but negative staining for CD20,CD68 and CD30.The patient was diagnosed as thymoma-associated graftversus-host-like disease,myasthenia gravis and mucosal candidiasis.After treatment with tacrolimus and prednisone,the condition was gradually relieved.

Objective　To prevent the after cataract induced by lens epithelial cells proliferation from postoperative cataract, monoclonal antibody (McAb) specific for rabbit lens epithelial cell is made, it will be the carrier further to be conjugated with cytotoxin. The conjugations will inhibit lens epithelial cells growth and not damage the other tissues of eye. Thereby McAb is the experimental bases of preventing after cataract.Methods　BALB/c mice were immunized by mixture of rabbit lens epithelial cells and Freund's adjuant. The immunized mouse spleen cells were fused with parental mouse myeloma cells (BALB/c　SP2/0) using polyethylene glycol (PEG-4000). The fused cells were selectively cultured by hypoxanthine, aminopterin and thymidine (HAT) culture medium. The specificities of the supernatant from hybridomas were tested by indirect immunofluorescence and immunohistochemistry SP (Streptavidin peroxdase conjugated method). The positive hybridomas were further cloned three times by methylcellulose culture medium to ensure monoclonality. At last, the consensual reaction of McAb was tested on human eye tissues.Results　Hybridomas were produced by fusion of spleen cells of immunized mice and mouse myoloma cells (SP2/0) with PEG-4000, and grown selectively in medium containing HAT after 16 days. Antibodies of the supernatant from hybridomas were tested on frozen sections of rabbit lens epithelial cell by indirect immunofluorescence. Only a positive clone secreted McAb against antigen of rabbit lens epithelial cell. The specificity of McAb was tested on paraffin sections of whole rabbit eye and whole human eye by immunohistochemistry SP. The results indicated that McAb was only positive to rabbit lens epithelial cell membrane and it was negative to the other tissues of rabbit or whole human eye tissues.Conclusions　McAb specific for rabbit lens epithelial cell was manufactured successfully. The specificity of McAb was strong. There was no consensual reaction on human eye tissues. It might be the experimental bases of further targeting chemotherapy on after cataract.

BACKGROUND:Bone nonunion may occur after limb trauma fracture. Internal fixation of implant is a common mode of repair, but fixation of different implants has different effects. OBJECTIVE:To explore the application value of different implant fixation in limb trauma of nonunion after femoral shaft fracture. METHODS:A retrospective analysis was performed on 72 cases of nonunion after femoral shaft fractures in Bozhou People’s Hospital from November 2012 to November 2013. They were divided into the observation group (36 cases) and control group (36 cases) according to the way of treatment, which were given interlocking intramedul ary nail and dynamic compression plate fixation. Length of incision, intraoperative blood transfusion volume, postoperative drainage volume, operation time, fracture healing time and functional recovery of knee joint were observed and compared between the two groups. RESULTS AND CONCLUSION:There was no statistical significance in operation time and length of the incision between the two groups (both P>0.05). Intraoperative amount of blood transfusion and postoperative drainage were significantly higher in the control group than in the observation group;the fracture healing time was significantly longer in the control group than in the observation group;infection rate in final fol ow-up was significantly higher in the control group than in the observation group (al P<0.05). No significant differences in preoperative knee joint International Knee Documentation Committee knee evaluation form and Lysholm score were found between the two groups (al P>0.05), but above two scores were significantly higher in the observation group than in the control group in final fol ow-up (al P<0.05). These findings suggest that compared with the dynamic compression plate, interlocking intramedul ary nail in treatment of bone nonunion after femoral shaft fracture can obtain good effect, firm fixation, low infection rate, and is more in line with the physiological and biomechanical requirements.

Background Vitreous hemorrhage in long-term produces toxic substances and influent the metabolism of eye tissue.Lens capsule is found more thin and transparent in the eyes with chronic vitreous hemorrhage.To research the effect of vitreous hemorrhage to lens is very important for the choose of the phaco operative timing.Objective The aim of this work was to investigate the ultrastructural change of lens epithelial cells(LECs) in the eyes with experimental vitreous hemorrhage.Methods The autologous blood of 0.1 ml was intravitreally injected in the left eyes of 8 general New Zealand white rabbits,and the equal amount of phosphate buffered saline(pH7.4) was used at the same way in the right eyes.Vitreous and fundus were examined with direct ophthalmoscope on 1,3,5,9,15,20,25,30 days to assess the inflammatory response after intravitreal injection.The specimens of lens anterior capsule were obtained in 30 days after injection and the ultrastructure and apoptosis of LECs were evaluated under the transmission electron microscope. Results No obvious ocular inflammatory response was seen throughout the experimental duration,and there was no vitreous hemorrhage in the right eyes after intravitreal injection.The vitreous hemorrhage agglutinated with clear boundary in the left eyes on 1 day after intravitreal injection,and the hemorrhage turned into dark-red color on the fifth day.On the fifteenth day after injection,the hemorrhage mass became to be grey color and the vitreous liquefaction occurred in the left eyes.The hemorrhage disappeared until 25 days.But in the one month after injection of self-blood,the vitreous showed the deeper red color.The early apoptosis appeared in the LECs of the left eyes in the thirty day,presenting the enlargement and broaden of intercellular space,the decrease of mitochondria number,vacuolar change,expanse of endoplasmic reticulum and disappearance of the nuclear membrane structure. Conclusions Vitreous hemorrhage leads to the ultrastructural pathological changes of lens.

Objective To investigate the treatment effects of Kansui root on severe acute pancreatitis (SAP) with abdominal compartment syndrome (ACS).Methods 16 cases of SAP were randomly divided into kansui root treatment group and control group according to random number table.Patients in control group received routine treatment including fasting, anti-shock, antibiotics and nutritional support.And the patients in kansui root group received routine treatment plus kansui root therapy.The clinical and laboratory parameters were determined and compared between the two groups.Results The relieving time of abdominal pain, bowel sound, the recovery time of hyperamylasemia, body temperature and leukocyte count in treatment group was (7.6±2.3)d, (6.1 ±3.1)d, (5.9±3.3)d, (5.2 ±3.2) d, (6.3 ±2.1)d, which were significantly shorter than those in control group [ ( 11.7 ± 2.1 ) d, ( 11.2 ± 2.3d, ( 10.2 ± 2.7) d, (9.2 ± 3.5 ) d, ( 11.1 ±3.3)d, P＜0.01 ) ].At the 3rd, 4th and 5th day, the intra-abdominal pressure in treatment group were also significantly lower than those in control group[ ( 19.8 ±3.1 )cmH2O vs(23.7 ±2.9) cmH2O, ( 12.3 ±2.7) cmH2O vs (21.3±1.5)cmH2O,(8.2±3.1)cmH2O vs (17.3 ±2.3)cmH2O,P＜0.05].Conclusions Severe acute pancreatitis has close relationship with Jiexiong syndrome in traditional chinese medicine.Kansui root is an effective therapy for alleviating high intra-abdominal pressure.

OBJECTIVETo investigate clinical effects of three-column reconstruction via single posterior approach for the treatment of unstable thoracolumbar fractures accompanied by posterior column injury.

METHODSFrom December 2008 to May 2010,three-column reconstruction via posterior approach was implemented to 21 patients with unstable thoracolumbar fractures accompanied by posterior column injuries. There were 13 males and 8 females, ranging in age from 23 to 54 years old(averaged,35.5 years old). Injured vertebrae: 1 patient had injury in T11, 4 patients had injuries in T12, 8 patients had injuries in L1, 5 patients had injuries in L2, 3 patients had injuries in L3. The Cobb angle was (25.34 +/- 3.42) degrees. The operation time,blood loss during operation, Cobb angle and the bony fusion were observed.

RESULTSTwenty-one patients were followed up, and the duration ranged from 24 to 27 years old, with an average of 25.6 months. The operation time ranged from 135 to 275 min, with a mean of 185 min. The blood loss during operation ranged from 700 to 1 650 ml (averaged, 870 ml). All the patients had complete decompression. Postoperative Cobb angle was (4.01 +/- 2.03) degrees, and (4.34 +/- 2.38) degrees at the latest follow-up. All the patients got bony fusion.

CONCLUSIONTo the patients with unstable thoracolumbar fractures accompanied by posterior column injuries, three-column reconstruction via single posterior approach has both anterior approach and posterior approach advantages, which can obtain excellent clinical outcomes.

Adult ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; adverse effects ; methods ; Spinal Fractures ; diagnostic imaging ; surgery ; Thoracic Vertebrae ; injuries ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

Metabolic engineering provide powerful tools for the systematic manipulation of cellular metabolic activities. The ptsG gene for glucose-specific transporter Enzyme II CBGlc of the phosphotransferase system was knock-out so as to reduce the accumulation of acetic acid in the high cell-density culture of Escherichia coli on excess glucose. The chloramphenicol-resistant cassette with short shared sequences on both ends generated by PCR was electroporated into Escherichia coli DH5alpha and JM109. Recombination between linear DNA cassettes and Escherichia coli chromosomes took place by Red recombinase functions. Therefore, the ptsG gene was disrupted to construct the mutants called DH5alphaP and JM109P. There was no difference between the mutants and parent strains in LB media.However, in LB media supplemented with glucose, the mutants of Escherichia coli deficient in ptsG showed greater biomass, together with exploiting more glucose. The maximal cell density obtained with DH5alphaP was approximately 3 times more than that of DH5alpha, then the result of JM109P increased fourfold. The products of recombinant protein TNF respectively accounted for 24.3% of total cellular protein in DH5alphaP with A600 8.28 and 20.8% of total cellular protein in JM109P with A600 7.62. The specific volume expression amount of TNF was greater in the ptsG mutant than in its parent strain. These results demonstrate that the ptsG-mutant strains will be available for high cell-density culture.
Culture Media ; Escherichia coli ; genetics ; growth & development ; Fermentation ; Mutation ; Phosphoenolpyruvate Sugar Phosphotransferase System ; genetics ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Tumor Necrosis Factor-alpha ; biosynthesis