Objective To study the method of the cultured human conjunctival epithelium combined with amniotic membrane transplantation for treatment of conjunctival defect. Methods 20 eyes of 10 rabbits were divided into study group (10 right eyes ) and contoal group (10 left eyes ). The conjunctival epithelium got from conjunctiva of fornix of study group about 1mm?1mm was explanted on human amniotic membrane and was cultured in vitro by tissue inoculation 2 weeks before operation. The bulbar conjunctivals of 20 eyes were excised. The conjunctival defect was covered by the graft in the study group and uncovered in the control group. The growth features of the grafts were observed. Results The conjunctival epithelium explanted on human amniotic membrane connected each other into membrane in 6 days, and multi-layer cells were observed. In the study group, all grafts were inosculated with and were similar to normal conjunctival in 2 weeks. There was not immune rejection occurred. In the control group, the conjunctival defect was recovered by scar. Conclusion It is effective of the cultured conjunctival epithelium combined with amniotic membrane transplantation for treatment of conjunctival defect.

Objective To study the culture method of conjunctival epithelium in vitro.Methods The rabbit conjunctival epithelium was cultured by tissue inoculation and was explanted in culture dish or on human amnionic membrane for fwo weeks.The cell morphology and growth features were observed.Results The rabbit conjunctival epithelium explanted in culture dish connected each other into membrane at eighth days around,which were monolayer cells was observed.The rabbit conjunctival epithelium explanted on human amnionic membrane connected each other into membrane at sixth day,and multi-layer cells were observed.All cultured cells showed positive in CK13 staining by indirect immunofluorescence staining.Conclusions Tissue inoculation is a good approach of conjunctival epithelium culture,and multi-layer cells can be gained if the tissue be exlpanted on human amnionic membrane.

[Objective]To explore the treatment of intertrochanteric fracture with expandable proximal femoral nail and prevention and cure of some related complications.[Method]Fifty-eight cases of closed intertrochanteric fracture,including 28 male cases and 30 female,average 73.2 years old(range from 42 to 88)accepted treatment.The type of fracture according to Evans classification:5 cases were type Ⅰ,7 were type Ⅱ,28 were type Ⅲ,13 were type Ⅳ,and 5 were type Ⅴ.All cases were treated with expandable proximal femoral intramedullary nail after close reduction.[Result]Thirty-eight cases were followed up,from 12 to 36 months,average 18.5 months,all of the cases got clinical healing after 8-10 weeks postoperatively.Hip Harris score average was 91.8.Some complications occurred in 6 cases:1 case had secondary fracture nearby the greater trochanter needling insertion point,1 case femoral head peg was outside of the lock hole,and 1 case femoral head peg tip was broken during operation.And after operation,1 case had coxa vara deformation,1 case had femoral middle and superior segment fracture,and 1 case femoral head peg was exited.[Conclusion]Expandable proximal femoral intramedullary nail has some advantages as:minimal invasive,manipulation convenient,fixation reliability,remove easily,but surgeons must be familiar with the construction and fixation principle of the nail in the primary stage,and perform the operating instruction strictly,to reduce or avoid the complications occurrence because of mis-manipulation.

Ten adult rabbits (7 and ♀3; body weights 1.5～2 kg) were selected for thee present study. A solution of 2～10％ HRP (RZ=2.9) was injected into the subserosa of the caecum in seven rabbits and a solution of 5～10％ HRP into the deep peroneal nerve of Tsusanli (足三里) region in the other three. The uptake and retrograds transmission of HRP in the afferent fibres of both the somatic and visceral nerves were traced to the spinal ganglia. The range of segments where the neurons from which these two afferent fibres originate overlap each other. The results are a follows:1. Labelled sensory neurons from the region of the caecum where HRP was injected are observed in the spinal ganglia C_8～S_3 with a higher concentration in T_(11)～L_2.2. Labelled neurons from the region of Tsusanli are found in the spinal ganglia L_1～S_3 with a higher concentration in L_6～S_2.3. The ranges of distribution of labelled neurons from the two groups of afferent fibres overlap in the segments L_1～S_3.4. Most of the labelled cells are small and medium in size and the Iabelled cells are found more concentrated in the lumbosacral segments.

The afferent projections to the hippocampus were studied with HRP. Thirty three per cent solution of HRP was slowly injected into the dorsal or ventral hippocampus on its right side in 25 albino rats and HRP-labelled cells were found in the following nuclei:1. On the ipsilateral side: entorhinal cortex, medial septal nucleus, supramammillary nucleus, submammillothalamic nucleus, dorsal raphe nucleus, median raphe nucleus, locus coeruleus, diagonal band and nucleus of superior colliculus.2. On the contralateral side: areas CA_3 and CA_4 of hippocampus and locus coeruleus.

Objective To investigate the effect of auxiliary heterotopic partial liver transplantation in treating acute ischemic liver failure. Methods Auxiliary heterotopic partial liver transplantation was performed on pigs.The pigs were divided into two groups.Group I: The host′ liver was preserved in situ, liver artery was ligated, portal vein was constricted,and donor patial right liver was transplanted under the right lobe of the liver of the host.Portal venous blood supply was constructed only, and arterial blood supply was not constructed .Group B:Except both arterial blood supply and portal venous blood supply of the donor liver were constructed, and the other operative procedures were as the same as in Group A. The living condition, liver function, blood supply, pathology and bile secretion of the donor liver were observed. Results The survival rates in Group B before and after operation were higher than that in Group A .Serum bilirubin(SB) after the operation was higher than that before the operation in Group A, but showed no change in Group B.After the operation, SB in Group A was higher than that in Group B. In Group A, the donor liver didn't secret bile,and hepatocytes were necrosis.In Group B, the bile secretion and blood supply of donor liver were good,and hepatocyte of the donor liver were alive and proliferating actively.The hosts' liver was necrosis obviously in both groups. Conclusions Hosts' liver artery ligation and portal vein constriction can result in acute ischemic liver failure.Auxiliary heterotopic partial liver transplantation is effective in correcting liver failure.The good arterial blood supply must be constructed in the donor liver to get good donor liver quality.

Objective To compare the different goal-directed fluid therapy(GDFT)strategies on the recovery of tissue perfusion and postoperative recovery in patients undergoing hepatectomy. Methods Ninety patients undergoing hepatectomy with ASA physical status Ⅱor Ⅲ were divided into three groups based on fluid treatment by random number table method:control group(conventional fluid therapy), observation group Ⅰ(GDFT filled with colloi) and observation group Ⅱ(GDFT filled with crystalloids),with 30 cases in each group.The ScvO2,Lac and Glu were tested at 5 min before anesthesia induction(T1),5 min before hepatectomy(T2),5 min after hepatectomy(T3)and the end of operation(T4). The duration of operation,fluid requirement,urinary output, bleeding volume and the use of vasoactive agent were recorded. The exhaust time, ambulation time and postoperative hospital stay were recorded. Preoperative and postoperative liver and kidney function tests and postoperative complications were followed up.Results Compared with that in control group,the amount of crystalloids[(1408.5 ± 348.4) ml vs. (1 819.4 ± 315.1) ml],the amount of colloids [(468.6 ± 193.4) ml vs. (1 009.7 ± 440.9) ml],the total volume[(1 867.3 ± 370.4)ml vs.(2 821.3 ± 264.6)ml]were all lower,first flatus time[(51.8 ± 8.5)h vs.(63.6 ± 9.2) h], ambulation time [(3.4 ± 0.7) d vs. (4.3 ± 0.7) d] and postoperative hospital stay [(7.8 ± 1.7)d vs.(10.5 ± 2.9)d]were all shorter,ScvO2at T3,T4was higher,Lac at T2-T4were lower,Glu at T3,T4was lower, and ALT and AST on the third day and the fifth day after surgery were lower in observation groupⅠ(P<0.05).Compared with those in observation groupⅠ,the amount of crystalloids [(2 014.7 ± 388.2)ml vs.(1 408.5 ± 348.4)ml]was increased,and the incidences of postoperative nausea and vomiting[33.3%(10/30)vs.10.0%(3/30)]were significantly higher than those in observation groupⅡ(P<0.05). Conclusions GDFT using colloids in hepatectomy probably improves the microcirculation and tissue oxygenation, protects liver function, promotes gastrointestinal function recovery and shortens postoperative hospital stay. It has a much lower incidence of postoperative nausea and vomiting, compared with crystalloids.

OBJECTIVETo investigate the mutation characteristics of ATM gene in Chinese patients with ataxia-telangiectasia (AT).

METHODSMutation of ATM gene was screened by polymerase chain reaction, reverse transcription-polymerase chain reaction, polyacrylamide gel electrophoresis combined with DNA direct sequencing in two Chinese AT patients.

RESULTSA missense mutation of 1346(G>C) in exon 11, which was a homozygotic mutation, was identified in one patient; a nonsense mutation of 610 (G>T) in exon 6 combined with a missense mutation of 6679 (C>T) in exon 47, which was a compound heterozygotic mutation, were identified in the other patient. They were co-segregated with the disease and were localized within the functional domain of ATM gene.

CONCLUSIONTotally three novel ATM gene mutations were identified in two Chinese AT patients.

Ataxia Telangiectasia Mutated Proteins ; Base Sequence ; Cell Cycle Proteins ; genetics ; Child ; China ; Codon, Nonsense ; DNA Mutational Analysis ; DNA-Binding Proteins ; genetics ; Female ; Gene Frequency ; Humans ; Male ; Mutation ; Mutation, Missense ; Protein-Serine-Threonine Kinases ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Spinocerebellar Ataxias ; genetics ; Tumor Suppressor Proteins ; genetics

Objective To analyze the clinical manifestation and therapeutic method in patients with acute mushroom poisoning. Methods A retrospective study was conducted. The clinical data of 48 patients with acute mushroom poisoning admitted to Department of Poisoning Treatment of the 307th Hospital of PLA from January 2016 to May 2017 were analyzed. The clinical data including gender, age, clinical symptoms, onset season, initial symptoms, incubation time, the length of hospital stay, treatment, and prognosis. In addition to the conventional treatment, the patients with severe liver damage were treated with continuous blood purification (CBP). The changes in routine blood test, biochemical parameters, blood ammonia and coagulation function before and 1, 3 and 7 days after CBP were observed. Results There were 29 of male (60.4%) and 19 of female (39.6%) in 48 patients with acute mushroom poisoning, with an average age of (48.10±13.14) years. There were 9 patients suffering from gastroenteritis type, 26 suffering from liver damage type, 8 suffering from neuro-psychosis type, 2 suffering from hemolytic type, and 3 suffering from renal damage type. All of the poisoned patients had evident seasonal characteristic, mainly concentrated in the autumn, especially in August, according for 66.7% (32/48). The initial symptoms of poisoning patients were mainly manifested as nausea and vomiting (50.0%). In five kinds of poisoned patients, the incubation time [(1.44±1.15) hours] and the length of hospital stay [(3.50±2.33) days] of neuro-psychosis type was the shortest, and the incubation time of liver-damaged type [(10.63±3.50) hours] and the length of hospital stay of renal damage type [(20.67±0.58) days] was the longest. Patients received symptomatic treatment according to different types, among whom 12 patients with severe liver damage received additional treatment for CBP. After the treatment, alanine aminotransferase (ALT), aspartate aminotransferase (AST), MB isoenzyme of creatine kinase (CK-MB), lactate dehydrogenase (LDH), and prothrombin activity (PTA) were significantly improved as compared with those before CBP treatment, with significant differences between 7 days after CBP and before CBP [ALT (U/L): 213.08±127.30 vs. 2 766.83±1 909.66, AST (U/L): 50.00 (41.00, 85.00) vs. 2 142.00 (1 225.00, 3 126.00), CK-MB (U/L): 24.09±8.87 vs. 44.75±22.09, LDH (μmol·s-1·L-1):3.70±1.46 vs. 13.03±12.77, PTA: (79.08±24.29)% vs. (35.25±19.85)%, all P < 0.01]. Among 48 patients, 47 were cured and discharged, and 1 patient with liver failure died due to aggravation of liver dysfunction, abnormal coagulation and bleeding, and massive hemorrhage of gastrointestinal tract. Conclusions Acute mushroom poisoning patients demonstrated obvious seasonal characteristics, mostly liver-damaged type, and its initial symptoms were mainly presented as nausea, vomiting and other gastrointestinal manifestations. Early clarification of diagnosis, timely treatment, as well as providence with CBP treatment in severe patients should be carried out as soon as possible. In such a way the curative effect can be enhanced, the mortality can be reduced, and the prognosis of the patients could be improved.

Objective: To investigate the efficacy of prussian blue (PB) or its combination with hemoperfusion (HP) in the treatment of acute thallium poisoning. Methods: Forty-seven patients with acute thallium poisoning with complete data hospitalized in the 307th Hospital of PLA from September 2002 to December 2017 were enrolled, and they were divided into mild poisoning group (blood thallium < 150 μg/L, urinary thallium < 1 000 μg/L) and moderate-severe poisoning group (blood thallium ≥ 150 μg/L, urinary thallium ≥ 1 000 μg/L) according to the toxic degrees. All patients were given symptomatic supportive treatments such as potassium supplementation, catharsis, vital organ protections, neurotrophic drugs, and circulation support. The mild poisoning patients were given PB with an oral dose of 250 mg·kg^-1·d^-1, while moderate-severe poisoning patients were given PB combined HP continued 2-4 hours each time. The PB dose or frequency of HP application was adjusted according to the monitoring results of blood and urine thallium. Data of gender, age, pain grading (numeric rating scale NRS), clinical manifestations, blood and urine thallium before and after treatment, length of hospitalization and prognosis were collected. Results: Of the 47 patients, patients with incomplete blood and urine test results, and used non-single HP treatment such as plasmapheresis and hemodialysis for treatment were excluded, and a total of 29 patients were enrolled in the analysis. ①Among 29 patients, there were 20 males and 9 females, median age of 40.0 (34.0, 49.0) years old; the main clinical manifestations were nervous system and alopecia, some patients had digestive system symptoms. There were 13 patients (44.8%) in the mild poisoning group with painless (grade 0) or mild pain (grade 1-3) with mild clinical symptoms, the length of hospitalization was 17.0 (14.2, 21.5) days. There were 16 patients (55.2%) in the moderate-severe poisoning group with moderate pain (grade 4-6) or severe pain (grade 7-10) with severe clinical symptoms, the length of hospitalization was 24.0 (18.0, 29.0) days. ② After treatment, the thallium concentrations in blood and urine in the mild poisoning group were significantly lower than those before treatment [μg/L: blood thallium was 0.80 (0, 8.83) vs. 60.00 (40.00, 120.00), urine thallium was 11.30 (0, 70.10) vs. 370.00 (168.30, 610.00), both P < 0.01], the thallium concentrations in blood and urine in the moderate-severe poisoning group were also significantly lower than those before treatment [μg/L: blood thallium was 6.95 (0, 50.50) vs. 614.50 (245.00, 922.00), urinary thallium was 20.70 (1.95, 283.00) vs. 5 434.00 (4 077.20, 10 273.00), both P < 0.01]. None of the 29 patients died, and their clinical symptoms were improved significantly. All the 27 patients had good prognosis without sequela in half a year follow-up, and 2 patients with severe acute thallium poisoning suffered from nervous system injury. Conclusion In the acute thallium poisoning patients, on the basis of general treatment, additional PB in mild poisoning group and PB combined with HP in moderate-severe poisoning group can obtain satisfactory curative effects.