Volume 110  ·   Issue 05  ·  1997

OBJECTIVE To determine whether human cytomegalovirus (CMV) matrix protein PP150 is efficiently presented for CMV-specific CTL recognition. METHODS Recombinant vaccinia virus (Vac. PP150) encoding human CMV structural matrix protein PP150 was constructed with vaccinia vector PSC11 and it was used to stimulate peripheral blond mononuclear cells from 5 CMV seropositive individuals. RESULTS PP150-specific CTLs could be generated in all of them, which not only lysed Vac. PP150-infected fibroblasts, but also lysed CMV-infected targets. In the presence of RNA synthesis inhibitor Actinomycin D (Act D) or at very early stage of infection, PP150-specific CTL lysed CMV-infected targets as efficiently as in the absence of Act D or at late stage of infection. CONCLUSIONS PP150 exogenously introduced with the virus infection could be efficiently presented prior to viral DNA replication and PP150 is one of the major target antigens for CTL recognition.

OBJECTIVE To investigate the histologic and histogenetic relationship between various types of rhabdomyosarcomas (RMS) and embryonal skeletal muscle (ESM) for further understanding of the histogenesis, classification and pattern of differentiation of RMS. METHODS Fifty cases of variant types of RMS and 20 cases of ESM at different gestational ages were available. All specimens were stained with HE, PAS, Van Gieson, Masson, phosphotungstic acid hematoxilin and with antibodies for the demonstration of vimentin, desmin, HHF-35 and myoglobin by ABC method. RESULTS The results showed that the order of positive expression and the intensity of positive reaction of the different immunohistochemical staining were consistent with the degree of differentiation of the tumor and the development of the ESM. It is obvious that each type of RMS is composed of tumor cells in different degree of differentiation and is derived from primitive mesodermal cells which are capable of potential differentiation towards mature skeletal muscles. CONCLUSIONS Based on the results of this study, an ideal histologic classification of RMS should reflect not only the cell morphology and histologic structures but also the degree of differentiation of the tumor cells.

OBJECTIVE To investigate the relationship between exogenous estrogen and breast cancer risk. METHODS Female rats were randomly divided into three groups, namely diethylstilbestrol (DES), norethindrone compositae (CoNET) and control group. The histological structure and ultrastructural changes of mammae were observed. The levels of sexual hormones in serum were determined and the AgNOR counts, DNA contents and steroid receptor contents in mammary epithelium were also detected. RESULTS In DES group, the level of progesterone (10.38 ng/ml) was obviously lower than that in the control group (13.37 ng/ml); the incidence of hyperplasia of mammary gland (73.33%) was significantly higher than that in the control group (7.69%); and the degree of hyperplasia was obviously more serious than that in the control group. Moreover, there were 13.33% of rats with atypical hyperplasia in DES group. The DNA contents, AgNOR counts and estrogen receptor (ER) positive rate were markedly higher in DES group (95.60, 2.43 and 71.71% respectively) than in the control group (83.07, 1.88 and 40% respectively). However, in CoNET group, there were no obvious influences on ER, AgNOR and DNA in mammary epithelium. CONCLUSIONS Exogenous estrogen (DES) could affect the levels of sexual hormones in serum, accelerate the DNA duplication, increase the AgNOR counts and ER contents, and induce atypical hyperplasia and ultrastructural changes of mammary gland, hence becoming a latent risk factor of breast cancer. However, the RESULTS failed to suggest that the contraceptive, CoNET, could increase the risk of breast cancer.

OBJECTIVE To determine the profile of lipidemia in patients with gallstones and the characteristics of lipidemia in different kinds of gallstone in gallbladder for predicting high risk subjects predisposed to gallstone formation by means of some serum parameters. METHODS Serum lipids and apolipoproteins levels in 47 patients with stone in gallbladder (stone group) were compared with those in 19 inpatients without stone (control group). The characteristics of lipidemia in different kinds of gallbladder stones were also compared. RESULTS Serum apolipoprotein (Apo) A1, C2 and E levels in the stone group significantly increased as compared with the control group (P<0.01), but there were no statistically significant differences in TC, TG, LDL-C, HDL-C, HDL-C/TC, LDL-C/HDL-C, Apo A2, B and C3 and Apo A1/Apo B levels between the stone and control groups (P>0.05). Increased levels of serum LDL-C and Apo B and the LDL-C/HDL-C ratio (P<0.05) were characterized as an index of cholesterol stones, while elevated levels of serum TG and Apo C2 (P<0.05) as an index of pigment stones. CONCLUSIONS Serum apolipoproteins might be more sensitive parameters as compared with serum lipids in distinguishing patients with stones from subjects without stones. There are different profiles of serum lipids and apolipoproteins in different chemical types of gallbladder stones.

OBJECTIVE To investigate the effect of electrochemical therapy (ECT) in the treatment of middle and late-stage lung cancers. METHODS Three hundred and eighty-six cases (287 males md 99 females aged 25-78, with an average age of 51 years) with middle and late-stage lung cancers were treated with ECT. Two hundred and three patients had squamous cell carcinoma, 138 adenocarcinoma and 45 undifferentiated cancer. Diameters of the cancers were respectively 4.0-6.0 cm in 153 cases, 6.1-8.0 cm in 82 cases, 8.1-10.0 cm in 102 cases and over 10.0 cm in 49 cases. In this group, none was at stage I, 103 cases were at stage Ⅱ, 89 cases at Ⅲa, 122 cases at Ⅲb and 72 cases Ⅳ. Of the 386 cases, 152 cases (39.4%) also suffered from hypertension, heart disease etc. Anode and cathode platinum electrodes were inserted accurately into the tumour mass. Distance between two electrodes was 2.0-2.5 cm. Electrodes were connected to a special ECT instrument. The current was maintained at 6-8 V and 80-100 mA. One hundred coulombs are used for treating 1 cm diameter of tumour mass. RESULTS Of the 386 cases, 99 cases (25.6%) achieved complete remission (CR). 179 cases (46.4%) achieved partial remission (PR), 59 cases (15.3%) had no change (NC) and 49 cases (12.7%) had progression of disease (PD). Effective rate CR + PR) was 72% (278 cases). One to five year survival rates were 86.3%, 76.4%, 58.8%, 39.9% and 29.5%, respectively. CONCLUSIONS ECT is easy to use. It is effective, safe, less traumatic and makes patients recover quickly. This is a new and effective method to treat patients with tumours who are inoperable and can not receive chemotherapy or radiotherapy.

OBJECTIVE To ascertain the prevalence and clinical features of mitochondrial tRNALeu(UUR) gene nucleotide (nt) 3243 A→G mutation diabetes mellitus in Chinese and to establish the approaches for genetic diagnosis of this subtype of diabetes during routine daily clinical practice. METHODS Mitochondrial nt 3243 A→G mutation was screened among 207 unrelated non-insulin-dependent diabetes mellitus (NIDDM) patients by using polymerase chain reaction (PCR)/Apa Ⅰ restriction endonuclease digestion. Samples with positive result were confirmed by DNA sequence analysis. Genetic and clinical analysis were carried out in family members of the proband with positive genetic diagnosis. RESULTS Positive results were found in two subjects during screening of unrelated NIDDM patients, which account for 2.4%-11.1% of NIDDM subjects with uncommon clinical features (with early age-of-onset, with low body-mass-index and on insulin therapy) or with positive family history of diabetes consistent with or unable to exclude from maternal inheritance. Genetic diagnosis was positive in 10 out of 25 family members in the pedigrees of these two probands. CONCLUSIONS Mitochondrial tRNALeu(UUR) gene nt 3242 A→G mutation should be considered in diabetic patients with the above-mentioned clinical features. The genetic diagnosis in daily clinical practice can be performed with allele specific priming amplification, or PCR/Hae Ⅲ or Apa Ⅰ digestion which are technically simple to perform and diagnostically easy to define.

OBJECTIVE To explore the distribution frequencies of vitamin D receptor (VDR) gene polymorphisms in Chinese and the relationship between VDR genotypes and bone mineral density (BMD) or serum osteocalcin level in Chinese women. METHODS Polymorphisms of VDR gene were analyzed by polymerase chain reaction (PCR) to amplify the DNA sequence and three restriction enzymes (namely BsmI, ApaI and TaqI) to digest the PCR products in 223 subjects. Some of the VDR genotypes were also confirmed by Southern hybridization analysis. BMD was measured at the spine and proximal femur by dual-energy X-ray absorptiometry (DEXA). Serum osteocalcin concentrations were determined by radioimmunoassay. RESULTS Chinese had high frequencies of "b,a,T" alleles, 95%, 75% and 95% respectively, which were much different from Caucasians. "BB" or "AA" genotype had tendency for higher BMD at some sites in the group of young women, whereas in the group of postmenopausal women, "bb" or "aa" genotype had relationship with lower BMD at femoral neck and trochanter. Furthermore, no relationship were found between VDR genotypes and the serum osteocalcin level in Chinese women. CONCLUSIONS Allele frequencies of VDR gene in Chinese are different from those in Caucasians. VDR gene polymorphisms are associated with BMD in Chinese, but in a different pattern from other reports. Further study on predicting function of VDR for osteoporosis is necessary.

OBJECTIVE To detect the cDNA sequence of immunoreactive growth hormone (irGH) in human lymphocytes and to compare it with that of human growth hormone (hGH) in the pituitary. METHODS The mRNA of irGH in human peripheral blood mononuclear cells (PBMC) was detected by Northern blot analysis. The cDNA of irGH was obtained by reverse-transcriptase and polymerase chain reaction amplification and obtained by screening from a cDNA library of human T lymphocytes. The sequence of irGH cDNA was determined and compared with that of the pituitary hGH. RESULTS Very low level of irGH mRNA was detected in human PBMCs by Northern blot analysis. The cDNA sequence of the entire open reading frame of human irGH was 651 bp, which is identical to that of pituitary hGH. CONCLUSIONS The same hGH molecule can be produced by human pituitary somatotrophic cells and human peripheral lymphocytes.

OBJECTIVE To study the distribution of nitric oxide synthase (NOS) in the enteric nervous system of children with cardiac achalasia. METHODS Biopsy specimens of the lower esophagus, cardia, gastric fundus and pylorus from 13 patients with cardiac achalasia and 6 controls were obtained and studied histochemically with nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) and immunohistochemically with a specific polyclonal antiserum. RESULTS NOS was abundant in the myenteric plexus and the nerve fibers of musculatures in the esophagus, cardia and gastric fundus in control group, while it was nearly absent in the patient group. The distribution of NOS in the pylorus was similar in the two groups. CONCLUSIONS These findings suggest that a lack of NOS in the lower esophagus, cardia and gastric fundus is involved in the pathophysiology of cardiac achalasia in children.

OBJECTIVE To investigate the anatomical and histological changes of the lengthened gut by a special intraluminal tubular expander on the basis of the theory of "tension-stress". METHODS Nineteen New Zealand white rabbits were used. A 5 cm segment of the terminal ileum with a pedicle was put underneath the abdominal wall, with its proximal end closed and its distal end exteriorized as a stoma. One week later, as the wound healed well, an expander made of a small rubber tube was inserted into the short gut loop through the stoma. Three milliliters of water was injected as an initial dose. Then 0.5 ml water as an increment was injected subsequently every 12 hours until the total amount of water reached 15 ml and kept stable for two weeks. RESULTS Anatomically, the length and the capacity of the intestinal loop were recorded, after 15 ml expanding, as an average of 150% lengthening of the original and an average of 293% expansion. Two weeks after the removal of the expander, partial shrinking was observed. A stable lengthening had 123% of the original length, while the capacity reduced to 200% of the original one. Histologically, there was no remarkable changes of the mucosa or submucosa. But significant hypertrophy of the musculature and serosa layer was observed. The total thickness of the musculatures was 618% of the normal control's. Ultrastructurally, there were enlargement of the smooth muscle cells, increase in number and size of the mitochondria in the cytoplasm and widening of the intercellular space. CONCLUSIONS A short gut can be lengthened by a tubular expander following the rule of gradual tension and stress.

OBJECTIVE To investigate the expression of proliferating cell nuclear antigen (PCNA) and its clinical significance in Wilms' tumor. METHODS PCNA immunoreactivity on paraffin-embedded tissues was assessed retrospectively in 35 cases using labelled streptaridin biotin (LSAB) procedure, and its association with clinical and other biological features was studied. RESULTS Twelve tumors (34.29%) expressed PCNA. Expression rate increased with the advance of stage (P<0.05). The S-phase fraction and proliferation index (PI) in children with PCNA positive expression were higher than those in children with negative PCNA expression (P<0.05). The survival rate (16.67%) in patients with positive PCNA expression was significantly lower than that (73.91%) in no-expression ones (P<0.01). CONCLUSIONS PCNA is a sensitive indicator for cell proliferation and is clinically useful in identifying a poor prognostic group of patients with Wilms' tumor.

OBJECTIVE To investigate whether or not estrogen receptor and other sex hormone receptors are present in hemangioma tissues of children. METHODS Fifty-two specimens of hemangiomas were taken from 52 children. The specimens were doubly stained with hematoxylin-eosin and enzyme-linking affinity immunohistochemistry. The prepared tissue slides were observed under the microscope to search for estrogen receptors (ER), progestogen receptors (PR) and androgen receptors (AR) in all the cells. RESULTS ER, PR and AR were detected in various kinds of hemangiomas. The mean positive cell rate of each sex hormone receptor in different hemangioma tissues revealed no significant distinction in statistics (P>0.05). CONCLUSIONS The results suggest that hemangiomas are one of the target tissues of estrogen. Estrogen, ER, PR, and AR may play an important role in the growth and development of hemangiomas.

OBJECTIVE To observe the developmental changes of the acetabulum after reduction of developmental dislocation of the hip and require evolutionary regularity of acetabular dysplasia. METHODS A follow-up in an average of 7.4 years was carried out in 117 patients (161 hips) with developmental dislocation of the hip after reduction. By the series of X-ray films, acetabular index, acetabular-head index, Sharp's angle, ACM angle (Idelberger's angle) and anteversion were observed. At the same time, centre-head distance discrepancy was measured. RESULTS Acetabular index returned to normal gradually as the time went by and significantly within one year after the reduction of developmental dislocation of the hip. The process of recovery was nearly stable three years later. Acetabular index in the dysplasia group was above 39 degrees before reduction and decreased slowly after the reduction. However, it was still up to 30 degrees after 3 years observation. The value of centre-head distance discrepancy decreased gradually as the years went by after reduction. CONCLUSIONS If acetabular index before reduction is above 39 degrees and is still up to 30 degrees three years after reduction, acetabular dysplasia can be diagnosed.

OBJECTIVE To determine whether circulating tumor cells are present in patients with localized or disseminated neuroblastoma (NB). METHODS Sixteen pediatric malignant tumor cell lines, including 10 NB, 4 primitive neuroectodermal tumor (PENT), 2 Wilms' tumor and 39 samples (tumor tissue, bone marrow and peripheral blood) from 8 patients with NB, were evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) using two markers: neuroendocrine protein gene products 9.5 (PGP 9.5) and tyrosine hydroxylase (TH). RESULTS Analysis of RT-PCR products by agarose electrophoresis demonstrated that in culture cell lines the specificity for NB was highest in TH (9/10 with NB, 3/4 with PNET, 0/2 with Wilms' tumor) and the sensitivity was highest in PGP 9.5 (10-7). PGP 9.5 and TH mRNA were undetectable in the peripheral blood mononuclear cells (PBMNC) from normal children. Expression of these markers in 8 patients with NB at diagnosis and during treatment was found positive in 5 out of 8, all had been proved paralleling with their disease status. Two of the 5 became negative after further treatment. CONCLUSIONS These systems allow the detection of circulating NB cells with higher sensitivity and specificity. When RT-PCR is clinically used for the detection of NB cells in blood, that both PGP 9.5 and TH should be selected is recommended. The assessment of bone marrow alone might not be used as a single index of tumor dissemination.

OBJECTIVE To evaluate regeneration and proliferation of the mucosa cells following intestinal ischemia/reperfusion (I/R) injury. METHODS Sixty rats (200-250 g) were examined after intestinal I/R injury with immunohistochemical (BrdU) method and flow cytometric analysis. RESULTS Severe damage to the villi was evident at 2 hours reperfusion and the rate of cell proliferation was lower (P<0.01). All the specimens has healed mucosa with re-epithelialized villi tips after 2 days regeneration, but the villi were scarce and tiny. After two and four days regeneration, the rate of cell proliferation was significantly higher than that of controls (P<0.01). CONCLUSIONS Our study suggests that the intestinal regeneration following I/R injury in rats occurs early. The immunohistochemical method with BrdU-labeled technique is a rapid method for quantifying the rate of cellular DNA synthesis.

OBJECTIVE To investigate the clinical manifestations and factors relating to prognosis in biliary atresia (BA). METHODS Twenty-four cases were reviewed on clinical manifestations and bile flow after operation. All patients underwent hepatic portoenterostomy. RESULTS Twenty-three cases (95%) got bile flow postoperatively. Sixteen (69%) of 23 patients had 27 episodes of cholangitis. There were significant differences (P<0.05) in the amount and bilirubin concentration of bile flow before and after cholangitis. Cholangitis also affected the life quality of BA patients. CONCLUSIONS We consider that early finding and prevention of cholangitis can improve the prognosis of BA patients.

OBJECTIVE To determine the pathological classification and the clinical characteristics of intestinal duplication. METHODS Eighty-four pediatric patients with intestinal duplication were proved on operation in the last 20 years in the Second Hospital of Harbin Medical University and Harbin Children's Hospital. The pathological classifications were as follows: enteral septum type (2), parietal cyst type (9), parenteral canal type (31), parenteral cyst type (35), and solitary type (1). The clinical presentations and complications included: abdominal mass (24), illius (41), hemorrhage of digestive tract (15) and intestinal perforation with peritonitis (3). Two asymptomatic cases were discovered incidentally. RESULTS Among the 84 patients, 82 healed, 1 newborn complicated by intestinal fistula died of septicemia, and the other one died of toxic shock. CONCLUSIONS Recognizing the pathological types, common presentations and complications, and applying the corresponding examination methods timely, doctors may improve the diagnostic accuracy.

OBJECTIVE To evaluate the effect of continuous balloon distension on the treatment of anorectal stenosis. METHODS Animal experiment on 17 guinea pigs and clinical study on 13 patients with anorectal stenosis were carried out. The procedure of the continuous balloon distension was followed to stimulate the growth of the anus or stenotic segment. RESULTS In animal experiment, the procedure induced muscle growth of the external anal sphincter at a rate of 2 mm per day. The grown muscle showed good contractile ability. By histological examination, it was found that the muscle cells were hypertrophic and Schwann's cells of the nerve fibers in the muscle became hyperplastic. Under electron-microscope, the muscle sarcomeres were found splitting both transversely and longitudinally. Twelve of the 13 patients were successfully treated by continuous balloon distension within 5 to 15 days (average: 6.7 days). During 4 to 14 months follow-up, all patients defecated without difficulty. CONCLUSIONS Continuous balloon distension may induce growth of the external anal sphincter. This procedure is an acceptable therapy for anorectal stenosis.

Advances in the field of molecular biology in medicine has provided new insights into the working of cells. In the recent 10 years, molecular endocrinology has radically transformed the understanding of endocrine physiology and pathophysiology.